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Fuchs B, Mert S, Kuhlmann C, Birt A, Hofmann D, Wiggenhauser PS, Giunta RE, Chavez MN, Nickelsen J, Schenck TL, Moellhoff N. In Vivo Biocompatibility of Synechococcus sp. PCC 7002-Integrated Scaffolds for Skin Regeneration. J Funct Biomater 2024; 15:295. [PMID: 39452593 PMCID: PMC11508603 DOI: 10.3390/jfb15100295] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2024] [Revised: 09/27/2024] [Accepted: 10/01/2024] [Indexed: 10/26/2024] Open
Abstract
Cyanobacteria, commonly known as blue-green algae, are prevalent in freshwater systems and have gained interest for their potential in medical applications, particularly in skin regeneration. Among these, Synechococcus sp. strain PCC 7002 stands out because of its rapid proliferation and capacity to be genetically modified to produce growth factors. This study investigates the safety of Synechococcus sp. PCC 7002 when used in scaffolds for skin regeneration, focusing on systemic inflammatory responses in a murine model. We evaluated the following three groups: scaffolds colonized with genetically engineered bacteria producing hyaluronic acid, scaffolds with wild-type bacteria, and control scaffolds without bacteria. After seven days, we assessed systemic inflammation by measuring changes in cytokine profiles and lymphatic organ sizes. The results showed no significant differences in spleen, thymus, and lymph node weights, indicating a lack of overt systemic toxicity. Blood cytokine analysis revealed elevated levels of IL-6 and IL-1β in scaffolds with bacteria, suggesting a systemic inflammatory response, while TNF-α levels remained unaffected. Proteome profiling identified distinct cytokine patterns associated with bacterial colonization, including elevated inflammatory proteins and products, indicative of acute inflammation. Conversely, control scaffolds exhibited protein profiles suggestive of a rejection response, characterized by increased levels of cytokines involved in T and B cell activation. Our findings suggest that Synechococcus sp. PCC 7002 does not appear to cause significant systemic toxicity, supporting its potential use in biomedical applications. Further research is necessary to explore the long-term effects and clinical implications of these responses.
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Affiliation(s)
- Benedikt Fuchs
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Sinan Mert
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Constanze Kuhlmann
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Alexandra Birt
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Daniel Hofmann
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Paul Severin Wiggenhauser
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Riccardo E. Giunta
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
| | - Myra N. Chavez
- Institute of Anatomy, University of Bern, 3012 Bern, Switzerland;
| | - Jörg Nickelsen
- Molecular Plant Science, Department Biology I, LMU Munich, 80336 Munich, Germany;
| | | | - Nicholas Moellhoff
- Division of Hand, Plastic and Aesthetic Surgery, LMU University Hospital, LMU Munich, 80336 Munich, Germany; (S.M.); (C.K.); (A.B.); (D.H.); (P.S.W.); (R.E.G.); (N.M.)
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Yang Y, Zheng S, Chu H, Du C, Chen M, Emran MY, Chen J, Yang F, Tian L. Subchronic Microcystin-LR Aggravates Colorectal Inflammatory Response and Barrier Disruption via Raf/ERK Signaling Pathway in Obese Mice. Toxins (Basel) 2023; 15:toxins15040262. [PMID: 37104200 PMCID: PMC10145857 DOI: 10.3390/toxins15040262] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/31/2022] [Revised: 03/03/2023] [Accepted: 03/14/2023] [Indexed: 04/05/2023] Open
Abstract
Microcystin-LR (MC-LR) is an extremely poisonous cyanotoxin that poses a threat to ecosystems and human health. MC-LR has been reported as an enterotoxin. The objective of this study was to determine the effect and the mechanism of subchronic MC-LR toxicity on preexisting diet-induced colorectal damage. C57BL/6J mice were given either a regular diet or a high-fat diet (HFD) for 8 weeks. After 8 weeks of feeding, animals were supplied with vehicle or 120 μg/L MC-LR via drinking water for another 8 weeks, and their colorectal were stained with H&E to detect microstructural alterations. Compared with the CT group, the HFD and MC-LR + HFD-treatment group induced a significant weight gain in the mice. Histopathological findings showed that the HFD- and MC-LR + HFD-treatment groups caused epithelial barrier disruption and infiltration of inflammatory cells. The HFD- and MC-LR + HFD-treatment groups raised the levels of inflammation mediator factors and decreased the expression of tight junction-related factors compared to the CT group. The expression levels of p-Raf/Raf and p-ERK/ERK in the HFD- and MC-LR + HFD-treatment groups were significantly increased compared with the CT group. Additionally, treated with MC-LR + HFD, the colorectal injury was further aggravated compared with the HFD-treatment group. These findings suggest that by stimulating the Raf/ERK signaling pathway, MC-LR may cause colorectal inflammation and barrier disruption. This study suggests that MC-LR treatment may exacerbate the colorectal toxicity caused by an HFD. These findings offer unique insights into the consequences and harmful mechanisms of MC-LR and provide strategies for preventing and treating intestinal disorders.
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Affiliation(s)
- Yue Yang
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
| | - Shuilin Zheng
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, The Key Laboratory of Ecological Environment and Critical Human Diseases Prevention of Hunan Province, Department of Education, Hengyang Medical School, University of South China, Hengyang 421001, China
- Changsha Center for Disease Control and Prevention, Changsha 410004, China
| | - Hanyu Chu
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, The Key Laboratory of Ecological Environment and Critical Human Diseases Prevention of Hunan Province, Department of Education, Hengyang Medical School, University of South China, Hengyang 421001, China
| | - Can Du
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
| | - Mengshi Chen
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
| | - Mohammed Y. Emran
- National Institute for Materials Science (NIMS), 1-2-1 Sengen, Tsukuba 305-0047, Japan
| | - Jihua Chen
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
| | - Fei Yang
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha 410078, China
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, The Key Laboratory of Ecological Environment and Critical Human Diseases Prevention of Hunan Province, Department of Education, Hengyang Medical School, University of South China, Hengyang 421001, China
| | - Li Tian
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, China
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Yang Y, Wang H, Wang X, Chen L, Liu W, Cai D, Deng S, Chu H, Liu Y, Feng X, Chen J, Chen M, Wang C, Liu R, Pu Y, Ding Z, Cao D, Long D, Cao Y, Yang F. Long-term environmental levels of microcystin-LR exposure induces colorectal chronic inflammation, fibrosis and barrier disruption via CSF1R/Rap1b signaling pathway. JOURNAL OF HAZARDOUS MATERIALS 2022; 440:129793. [PMID: 36029734 DOI: 10.1016/j.jhazmat.2022.129793] [Citation(s) in RCA: 49] [Impact Index Per Article: 16.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/29/2022] [Revised: 07/20/2022] [Accepted: 08/15/2022] [Indexed: 02/05/2023]
Abstract
Microcystin-LR (MC-LR) is a very common toxic cyanotoxins threating ecosystems and the public health. This study aims to explore the long-term effects and potential toxicity mechanisms of MC-LR exposure at environmental levels on colorectal injury. We performed histopathological, biochemical indicator and multi-omics analyses in mice with low-dose MC-LR exposure for 12 months. Long-term environmental levels of MC-LR exposure caused epithelial barrier disruption, inflammatory cell infiltration and an increase of collagen fibers in mouse colorectum. Integrated proteotranscriptomics revealed differential expression of genes/proteins, including CSF1R, which were mainly involved in oxidative stress-induced premature senescence and inflammatory response. MC-LR induced chronic inflammation and fibrosis through oxidative stress and CSF1R/Rap1b signaling pathway were confirmed in cell models. We found for the first time that long-term environmental levels of MC-LR exposure caused colorectal chronic inflammation, fibrosis and barrier disruption via a novel CSF1R/Rap1b signaling pathway. Moreover, MC-LR changed the gut microbiota and microbial-related metabolites in a vicious cycle aggravating colorectal injury. These findings provide novel insights into the effects and toxic mechanisms of MC-LR and suggest strategies for the prevention and treatment of MC-caused intestinal diseases.
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Affiliation(s)
- Yue Yang
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China
| | - Hui Wang
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Xiaoyan Wang
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha, China
| | - Ling Chen
- First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, China
| | - Wenya Liu
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China
| | - Danping Cai
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Shuxiang Deng
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Hanyu Chu
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Ying Liu
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Xiangling Feng
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China
| | - Jihua Chen
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China
| | - Mengshi Chen
- Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China
| | - Chengkun Wang
- Department of Medical Pathology, School of Basic Medical, Hengyang Medical School, University of South China, Hengyang, China
| | - Ran Liu
- Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China
| | - Yuepu Pu
- Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China
| | - Zhen Ding
- Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, China
| | - Deliang Cao
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Dingxin Long
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Yi Cao
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China
| | - Fei Yang
- Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang, China; Hunan Provincial Key Laboratory of Clinical Epidemiology, Xiangya School of Public Health, Central South University, Changsha, China; Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China.
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Lipopolysaccharide from the Cyanobacterium Geitlerinema sp. Induces Neutrophil Infiltration and Lung Inflammation. Toxins (Basel) 2022; 14:toxins14040267. [PMID: 35448876 PMCID: PMC9024439 DOI: 10.3390/toxins14040267] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2022] [Revised: 04/04/2022] [Accepted: 04/06/2022] [Indexed: 11/17/2022] Open
Abstract
Glucocorticoid-resistant asthma, which predominates with neutrophils instead of eosinophils, is an increasing health concern. One potential source for the induction of neutrophil-predominant asthma is aerosolized lipopolysaccharide (LPS). Cyanobacteria have recently caused significant tidal blooms, and aerosolized cyanobacterial LPS has been detected near the cyanobacterial overgrowth. We hypothesized that cyanobacterial LPS contributes to lung inflammation by increasing factors that promote lung inflammation and neutrophil recruitment. To test this hypothesis, c57Bl/6 mice were exposed intranasally to LPS from the cyanobacterium member, Geitlerinema sp., in vivo to assess neutrophil infiltration and the production of pro-inflammatory cytokines and chemokines from the bronchoalveolar fluid by ELISA. Additionally, we exposed the airway epithelial cell line, A549, to Geitlerinema sp. LPS in vitro to confirm that airway epithelial cells were stimulated by this LPS to increase cytokine production and the expression of the adhesion molecule, ICAM-1. Our data demonstrate that Geitlerinema sp. LPS induces lung neutrophil infiltration, the production of pro-inflammatory cytokines such as Interleukin (IL)-6, Tumor necrosis factor-alpha, and Interferongamma as well as the chemokines IL-8 and RANTES. Additionally, we demonstrate that Geitlerinema sp. LPS directly activates airway epithelial cells to produce pro-inflammatory cytokines and the adhesion molecule, Intercellular Adhesion Molecule-1 (ICAM-1), in vitro using the airway epithelial cell line, A549. Based on our findings that use Geitlerinema sp. LPS as a model system, the data indicate that cyanobacteria LPS may contribute to the development of glucocorticoid-resistant asthma seen near water sources that contain high levels of cyanobacteria.
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Immunotoxic Effects Induced by Microcystins and Cylindrospermopsin: A Review. Toxins (Basel) 2021; 13:toxins13100711. [PMID: 34679003 PMCID: PMC8540411 DOI: 10.3390/toxins13100711] [Citation(s) in RCA: 25] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2021] [Revised: 10/01/2021] [Accepted: 10/06/2021] [Indexed: 12/17/2022] Open
Abstract
Cyanotoxin occurrence is gaining importance due to anthropogenic activities, climate change and eutrophication. Among them, Microcystins (MCs) and Cylindrospermopsin (CYN) are the most frequently studied due to their ubiquity and toxicity. Although MCs are primary classified as hepatotoxins and CYN as a cytotoxin, they have been shown to induce deleterious effects in a wide range of organs. However, their effects on the immune system are as yet scarcely investigated. Thus, to know the impact of cyanotoxins on the immune system, due to its importance in organisms’ homeostasis, is considered of interest. A review of the scientific literature dealing with the immunotoxicity of MCs and CYN has been performed, and both in vitro and in vivo studies have been considered. Results have confirmed the scarcity of reports on the topic, particularly for CYN. Decreased cell viability, apoptosis or altered functions of immune cells, and changed levels and mRNA expression of cytokines are among the most common effects reported. Underlying mechanisms, however, are still not yet fully elucidated. Further research is needed in order to have a full picture of cyanotoxin immunotoxicity.
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Arman T, Lynch KD, Montonye ML, Goedken M, Clarke JD. Sub-Chronic Microcystin-LR Liver Toxicity in Preexisting Diet-Induced Nonalcoholic Steatohepatitis in Rats. Toxins (Basel) 2019; 11:E398. [PMID: 31323923 PMCID: PMC6669744 DOI: 10.3390/toxins11070398] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/13/2019] [Revised: 07/02/2019] [Accepted: 07/04/2019] [Indexed: 02/07/2023] Open
Abstract
Microcystin-LR (MCLR) is a hepatotoxic cyanotoxin reported to cause a phenotype similar to nonalcoholic steatohepatitis (NASH). NASH is a common progressive liver disease that advances in severity due to exogenous stressors such as poor diet and toxicant exposure. Our objective was to determine how sub-chronic MCLR toxicity affects preexisting diet-induced NASH. Sprague-Dawley rats were fed one of three diets for 10 weeks: control, methionine and choline deficient (MCD), or high fat/high cholesterol (HFHC). After six weeks of diet, animals received vehicle, 10 µg/kg, or 30 µg/kg MCLR via intraperitoneal injection every other day for the final 4 weeks. Incidence and severity scoring of histopathology endpoints suggested that MCLR toxicity drove NASH to a less fatty and more fibrotic state. In general, expression of genes involved in de novo lipogenesis and fatty acid esterification were altered in favor of decreased steatosis. The higher MCLR dose increased expression of genes involved in fibrosis and inflammation in the control and HFHC groups. These data suggest MCLR toxicity in the context of preexisting NASH may drive the liver to a more severe phenotype that resembles burnt-out NASH.
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Affiliation(s)
- Tarana Arman
- Department of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USA
| | - Katherine D Lynch
- Department of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USA
| | - Michelle L Montonye
- Department of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USA
| | - Michael Goedken
- Department of Pharmacology and Toxicology, Rutgers University, Piscataway, NJ 08901, USA
| | - John D Clarke
- Department of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USA.
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Wu JX, Huang H, Yang L, Zhang XF, Zhang SS, Liu HH, Wang YQ, Yuan L, Cheng XM, Zhuang DG, Zhang HZ. Gastrointestinal toxicity induced by microcystins. World J Clin Cases 2018; 6:344-354. [PMID: 30283797 PMCID: PMC6163130 DOI: 10.12998/wjcc.v6.i10.344] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/23/2018] [Revised: 06/08/2018] [Accepted: 06/28/2018] [Indexed: 02/05/2023] Open
Abstract
Microcystins (MCs) are produced by certain bloom-forming cyanobacteria that can induce toxicity in various organs, including renal toxicity, reproductive toxicity, cardiotoxicity, and immunosuppressive effects. It has been a significant global environmental issue due to its harm to the aquatic environment and human health. Numerous investigators have demonstrated that MC exposure can induce a widespread epidemic of enterogastritis with symptoms similar to food poisoning in areas close to lakes. Both in vivo and in vitro studies have provided evidence of positive associations between MC exposure and gastrointestinal toxicity. The toxicity of MCs on the gastrointestinal tract is multidimensional. MCs can affect gastrointestinal barrier function and shift the structure of gut microbiota in different gut regions. Furthermore, MCs can inhibit the secretion of gastrointestinal digestive enzymes and the release of inflammatory cytokines, which affects the expression of immune-related genes in the intestine. The damage of the intestine is closely correlated to MC exposure because the intestine is the main site for the digestion and absorption of nutrients. The damage to the gastrointestinal tract due to MCs was summarized from different aspects, which can be used as a foundation for further exploration of molecular damage mechanisms.
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Affiliation(s)
- Jin-Xia Wu
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Hui Huang
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Lei Yang
- Department of Nutriology, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Xiao-Feng Zhang
- Department of Nutriology, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Shen-Shen Zhang
- Department of Nutriology, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Hao-Hao Liu
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Yue-Qin Wang
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Le Yuan
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Xue-Min Cheng
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Dong-Gang Zhuang
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
| | - Hui-Zhen Zhang
- Department of Environmental Hygiene, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan Province, China
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Ma J, Li Y, Duan H, Sivakumar R, Li X. Chronic exposure of nanomolar MC-LR caused oxidative stress and inflammatory responses in HepG2 cells. CHEMOSPHERE 2018; 192:305-317. [PMID: 29117589 DOI: 10.1016/j.chemosphere.2017.10.158] [Citation(s) in RCA: 40] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/26/2017] [Revised: 10/26/2017] [Accepted: 10/27/2017] [Indexed: 06/07/2023]
Abstract
Low dose but long-term exposure of microcystin-LR (MC-LR) could induce human hepatitis and promote liver cancer according to epidemiological investigation results, but the exact mechanism has not been completely elucidated. In the present study, a chronic toxicity test of MC-LR exposure on HepG2 cells at 0.1-30 nM for 83 d was conducted under laboratory conditions. The western blot assay result revealed that MC-LR entered HepG2 cells, even at the concentration of 0.1 nM, after 83 d of exposure, but no cytotoxicity was observed in the HepG2 cells, as determined by the CCK-8 and LDH tests. However, the results of the DCF fluorescence assay showed that the intracellular ROS level in the 30 nM MC-LR-treated cells was significantly higher than that of the control cells, and 5 and 10 nM of MC-LR exposure totally increased the activity of SOD in HepG2 cells. These results indicate that MC-LR exposure at low concentration also induced excessive ROS in HepG2 cells. Additionally, long-term exposure of MC-LR at low concentration remarkably promoted the expression of NF-κB p65, COX-2, iNOS, TNF-α, IL-1β, and IL-6 in the cells, suggesting that long-term MC-LR exposure at low concentration can induce inflammatory reaction to HepG2 cells, which might account for MC-induced human hepatitis. Thus, we hypothesized that the pathogenesis of human hepatitis and hepatocarcinoma caused by MCs might be closely associated with oxidative stress and inflammation.
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Affiliation(s)
- Junguo Ma
- College of Life Science, Henan Normal University, Xinxiang, Henan 453007, China
| | - Yuanyuan Li
- College of Life Science, Henan Normal University, Xinxiang, Henan 453007, China
| | - Hongying Duan
- College of Life Science, Henan Normal University, Xinxiang, Henan 453007, China
| | | | - Xiaoyu Li
- College of Life Science, Henan Normal University, Xinxiang, Henan 453007, China.
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Chen C, Liu W, Wang L, Li J, Chen Y, Jin J, Kawan A, Zhang X. Pathological damage and immunomodulatory effects of zebrafish exposed to microcystin-LR. Toxicon 2016; 118:13-20. [PMID: 27085306 DOI: 10.1016/j.toxicon.2016.04.030] [Citation(s) in RCA: 60] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2016] [Revised: 04/01/2016] [Accepted: 04/12/2016] [Indexed: 12/18/2022]
Abstract
Cyanobacterial blooms caused by water eutrophication have become a worldwide problem. Microcystins (MCs), especially microcystin-LR (MC-LR), released during cyanobacterial blooms exert great toxicity on fish and even lead to massive death. The present study mainly investigated the pathological damage and immune response of spleen, gut and gill in zebrafish exposed to MC-LR. Fish were exposed to 0, 1, 5 and 20 μg/L of MC-LR for 30 d. In zebrafish exposed to 5 and 20 μg/L MC-LR, edematous mitochondria, deformation of the nucleus and compaction of chromatin were observed in lymphocyte of spleen; frayed gut villi, exfoliation of epithelial cells and widespread cell lyses were observed in intestines; hyperemia in gill lamellae, epithelial tissue edema and uplift and lamellar fusion were observed in gill. Varied changed gene expression was observed in spleen, intestine and gill of zebrafish. The transcriptional levels of IFN-1 and IL-8 in spleen significantly up-regulated in 20 μg/L group, and the transcription of IL-1β and TNFα in spleen increased in 1 μg/L MC-LR treated fish. In addition, the mRNA levels of IFN-1, IL-1β, IL-8, TGF-β and TNF-α dramatically increased in intestine and gill in all MC-LR treated groups. The present studies indicated that MC-LR exposure caused marked pathological damage, however, fish could adjust actively the expression of innate immune-related genes to resist the tissue damage. Our findings provided strong evidence of the recovery potential of fish exposed to microcystins.
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Affiliation(s)
- Chuanyue Chen
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Wanjing Liu
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Li Wang
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Jian Li
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Yuanyuan Chen
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Jienan Jin
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Atufa Kawan
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China
| | - Xuezhen Zhang
- College of Fisheries, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, PR China.
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He J, Li G, Chen J, Lin J, Zeng C, Chen J, Deng J, Xie P. Prolonged exposure to low-dose microcystin induces nonalcoholic steatohepatitis in mice: a systems toxicology study. Arch Toxicol 2016; 91:465-480. [PMID: 26984711 DOI: 10.1007/s00204-016-1681-3] [Citation(s) in RCA: 65] [Impact Index Per Article: 7.2] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/26/2015] [Accepted: 02/09/2016] [Indexed: 12/20/2022]
Abstract
Microcystin-LR (MCLR), a cyanotoxin widely present in freshwater, has been shown to have potent acute hepatotoxicity. However, the chronic toxicity of low-dose MCLR remains confusing by traditional measurements of toxicity. This has impeded understanding of the chronic liver damage of low-dose MCLR and corresponding safety risks of the human exposure guideline value. Here, iTRAQ-based proteomics and NMR-based metabonomics were used to decipher the molecular toxicological signatures of low doses of MCLR in mice exposed to this agent for 90 days. Low levels of MCLR, even under the reported no observed adverse effect level, significantly altered hepatic protein expression, especially of proteins associated with lipid metabolism, transport, immune and proteolysis. Coherently, MCLR induced marked perturbations in lipid metabolites in both liver and serum. Integrated analysis of proteomic, metabolic, histological and cytokine profiles revealed that MCLR significantly inhibited fatty acid β-oxidation and hepatic lipoprotein secretion and promoted hepatic inflammation, resulting in nonalcoholic steatohepatitis disease (NASH). These findings for the first time provide compelling evidence that chronic exposure to low-level MCLR can induce NASH. These results also indicate that current guidelines for MCs in drinking water may be inadequate and associated with risks to human health.
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Affiliation(s)
- Jun He
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan, 430072, People's Republic of China
| | - Guangyu Li
- College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China
| | - Jun Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan, 430072, People's Republic of China
| | - Juan Lin
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan, 430072, People's Republic of China
| | - Cheng Zeng
- College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China
| | - Jing Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan, 430072, People's Republic of China
| | - Junliang Deng
- Shanghai Biotree Biotech Co., Ltd, Shanghai, 200433, People's Republic of China
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan, 430072, People's Republic of China.
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11
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Adamovsky O, Moosova Z, Pekarova M, Basu A, Babica P, Svihalkova Sindlerova L, Kubala L, Blaha L. Immunomodulatory Potency of Microcystin, an Important Water-Polluting Cyanobacterial Toxin. ENVIRONMENTAL SCIENCE & TECHNOLOGY 2015; 49:12457-12464. [PMID: 26380879 DOI: 10.1021/acs.est.5b02049] [Citation(s) in RCA: 43] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/05/2023]
Abstract
Microcystins (MCs) are primarily hepatotoxins produced by cyanobacteria and are responsible for intoxication in humans and animals. There are many incidents of chronic exposure to MCs, which have been attributed to the inappropriate treatment of water supplies or contaminated food. Using RAW 264.7 macrophages, we showed the potency of microcystin-LR (MC-LR) to stimulate production of pro-inflammatory cytokines (tumor necrosis factor α and interleukin-6) as a consequence of fast nuclear factor κB and nitrogen-activated protein kinase activation. In contrast to other studies, the observed effects were not attributed to the intracellular inhibition of protein phosphatases 1/2A due to lack of specific transmembrane transporters for MCs. However, the MC-LR-induced activation of macrophages was effectively inhibited by a specific peptide that blocks signaling of receptors, which play a pivotal role in the innate immune responses. Taken together, we showed for the first time that MC-LR could interfere with macrophage receptors that are responsible for triggering the above-mentioned signaling pathways. These findings provide an interesting mechanistic explanation of some adverse health outcomes associated with toxic cyanobacteria and MCs.
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Affiliation(s)
- Ondrej Adamovsky
- Faculty of Science, RECETOX, Masaryk University , Kamenice 753/5, 62500 Brno, Czech Republic
| | - Zdena Moosova
- Faculty of Science, RECETOX, Masaryk University , Kamenice 753/5, 62500 Brno, Czech Republic
| | - Michaela Pekarova
- Institute of Biophysics, Academy of Sciences , Královopolská 135, 612 65 Brno, Czech Republic
| | - Amrita Basu
- Faculty of Science, RECETOX, Masaryk University , Kamenice 753/5, 62500 Brno, Czech Republic
| | - Pavel Babica
- Faculty of Science, RECETOX, Masaryk University , Kamenice 753/5, 62500 Brno, Czech Republic
| | | | - Lukas Kubala
- Institute of Biophysics, Academy of Sciences , Královopolská 135, 612 65 Brno, Czech Republic
| | - Ludek Blaha
- Faculty of Science, RECETOX, Masaryk University , Kamenice 753/5, 62500 Brno, Czech Republic
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12
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Sedan D, Laguens M, Copparoni G, Aranda JO, Giannuzzi L, Marra CA, Andrinolo D. Hepatic and intestine alterations in mice after prolonged exposure to low oral doses of Microcystin-LR. Toxicon 2015. [PMID: 26210502 DOI: 10.1016/j.toxicon.2015.07.011] [Citation(s) in RCA: 47] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
Oral intake of Microcystin-LR (MC-LR) is the principal route of exposure to this toxin, with prolonged exposure leading to liver damage of unspecific symptomatology. The aim of the present paper was therefore to investigate the liver and intestine damage generated by prolonged oral exposure to low MC-LR doses (50 and 100 μg MC-LR/kg body weight, administrated every 48 h during a month) in a murine model. We found alterations in TBARS, SOD activity and glutathione content in liver and intestine of mice exposed to both doses of MC-LR. Furthermore, the presence of MC-LR was detected in both organs. We also found hepatic steatosis (3.6 ± 0.6% and 15.3 ± 1.6%) and a decrease in intraepithelial lymphocytes (28.7 ± 5.0% and 44.2 ± 8.7%) in intestine of 50- and 100-μg MC-LR/kg treated animals, respectively. This result could have important implications for mucosal immunity, since intraepithelial lymphocytes are the principal effectors of this system. Our results indicate that prolonged oral exposure at 50 μg MC-LR/kg every 48 h generates significant damage not only in liver but also in intestine. This finding calls for a re-appraisal of the currently accepted NOAEL (No Observed Adverse Effect Level), 40 μg MC-LR/kg body weight, used to derive the guideline value for MC-LR in drinking water.
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Affiliation(s)
- Daniela Sedan
- Área de Toxicología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP-CONICET), La Plata, Argentina.
| | - Martín Laguens
- Cátedra de Patología B, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, La Plata, Argentina.
| | - Guido Copparoni
- Área de Toxicología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP-CONICET), La Plata, Argentina.
| | - Jorge Oswaldo Aranda
- Área de Toxicología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP-CONICET), La Plata, Argentina.
| | - Leda Giannuzzi
- Área de Toxicología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP-CONICET), La Plata, Argentina; Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA-CONICET), La Plata, Argentina.
| | - Carlos Alberto Marra
- INIBIOLP (Instituto de Investigaciones Bioquímicas de La Plata), Cátedra de Bioquímica y Biología Molecular, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, La Plata, Argentina.
| | - Darío Andrinolo
- Área de Toxicología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP-CONICET), La Plata, Argentina.
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Li H, Cai Y, Xie P, Li G, Hao L, Xiong Q. Identification and expression profiles of IL-8 in bighead carp (Aristichthys nobilis) in response to microcystin-LR. ARCHIVES OF ENVIRONMENTAL CONTAMINATION AND TOXICOLOGY 2013; 65:537-45. [PMID: 23797979 PMCID: PMC3763166 DOI: 10.1007/s00244-013-9910-8] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 02/06/2013] [Accepted: 04/22/2013] [Indexed: 06/02/2023]
Abstract
Microcystin-LR (MCLR) is a widespread cyanotoxin and has immunotoxicity to animals, including fish. Chemokines are considered to play important roles in inflammatory response induced by MCLR. In this study, we cloned the full-length cDNA of interleukin-8 (IL-8) from bighead carp (Aristichthys nobilis) for the first time. The full-length IL-8 cDNA was 552 bp and contained a 297-bp open-reading frame that encoded for a 98-amino acid protein. The deduced IL-8 protein had a typical aspartic acid (D)-leucine (L)-arginine (R) and a CXC motif at the N-terminal, which were conserved in most fish species. Phylogenetic analysis showed that bighead carp IL-8 protein was grouped in the teleost IL-8 lineage 2. Under normal conditions, the expression of IL-8 is constitutive and weak in all tested tissues. However, MCLR treatment could significantly increase the transcription of IL-8 in bighead carp in a temporal- and dose-dependent pattern. The present study will help us to understand more about the evolution of IL-8 and its function in the MCLR induced proinflammatory response in bighead carp.
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Affiliation(s)
- Huiying Li
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
- Present Address: Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, CAS, Lumo Street, Wuhan, 430074 People’s Republic of China
| | - Yan Cai
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
| | - Guangyu Li
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
| | - Le Hao
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
| | - Qian Xiong
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072 People’s Republic of China
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14
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Rymuszka A, Adaszek Ł. Pro- and anti-inflammatory cytokine expression in carp blood and head kidney leukocytes exposed to cyanotoxin stress--an in vitro study. FISH & SHELLFISH IMMUNOLOGY 2012; 33:382-388. [PMID: 22641113 DOI: 10.1016/j.fsi.2012.05.021] [Citation(s) in RCA: 67] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/06/2012] [Revised: 05/11/2012] [Accepted: 05/14/2012] [Indexed: 06/01/2023]
Abstract
Cyanotoxins are toxic, secondary metabolites produced by different species of cyanobacteria that are present all over the world in aquatic environments. No data are available about the molecular mechanisms underlying the stress associated with exposure of fish immune cells to low concentrations of cyanotoxins. The purpose of this study was to determine whether the expression of cytokines that underlie immune regulation are changed after incubation of fish leukocytes with pure cyanotoxins: microcystin- LR (MC-LR), anatoxin-a (Antx-a), or an extract containing Antx-a. The study investigated the relative gene expression of four important cytokines, IL-1β, TNF-α, IL-10, and TGF-β, in carp head kidney and blood leukocytes exposed to toxins at concentrations of 0.01 or 0.1 μg/ml for 4 h. The data showed that pure toxins could induce dysregulation of pro-/anti-inflammatory cytokine expression. Expression of cytokine IL-1 β was highly upregulated following Antx-a exposure, whereas MC-LR induced merely moderate reactions. The expression of TNF-α mRNA was significantly suppressed in blood and head kidney cells incubated with toxins at the higher concentration. These results showed that pure toxins dysregulated the expression of pro-inflammatory cytokines IL-1β and TNF-α more promptly than the anti-inflammatory cytokines TGF-β and IL-10. In contrast, the studies demonstrated a clearly downward trend of pro-inflammatory cytokines and an upward trend of anti-inflammatory cytokines in leukocytes exposed to an extract containing defined concentrations of Antx-a. This study suggests that cyanotoxins present in aquatic environments may exert immunotoxic effects by altering the transcription of important mediators of the fish immune system.
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Affiliation(s)
- Anna Rymuszka
- The John Paul II Catholic University of Lublin, Institute of Biotechnology, Department of Physiology and Ecotoxicology, Lublin, Poland.
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15
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Yuan G, Xie P, Zhang X, Tang R, Gao Y, Li D, Li L. In vivo studies on the immunotoxic effects of microcystins on rabbit. ENVIRONMENTAL TOXICOLOGY 2012; 27:83-89. [PMID: 20549643 DOI: 10.1002/tox.20615] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/23/2009] [Revised: 03/22/2010] [Accepted: 03/27/2010] [Indexed: 05/29/2023]
Abstract
Microcystins (MCs) are the toxic molecules produced by common cyanobacterium in freshwater blooms. Their toxicities raise severe health issues in livestock and human beings. In current study, the immunotoxic effects of MC-LR were investigated in rabbit through evaluating the dynamics of white blood cell (WBC) numbers and cytokine production such as interleukin-3 (IL-3), IL-4, IL-6, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α). MCs at the high dose (50 μg MC-LReq kg(-1) ) significantly induced increase in the WBC number but decrease in the Th1 (IFN-γ, TNF-α) and Th2 (IL-3, IL-4, IL-6) production. In the low dose group(12.5 μg MC-LReq kg(-1) ), the number of WBC and the production of IFN-γ, IFN-α, IL-4, IL-3, and IL-6 increased gradually in first 12 h, reach the peaks at 12 h, and dropped after 24 h. Significantly positive correlations were found between the cytokines production of IL-4 and IL-6, IFN-γ and IFN-α, or IL-4 and IFN-γ. In conclusion, MC-LR is able to disturb the rabbit immune system and there exists time-dose response relationship in the MC-LR-eliciting perturbation, which probably give a better insight into investigating the immunotoxicity mechanisms of MCs in vivo. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012.
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Affiliation(s)
- Gailing Yuan
- Fisheries College of Huazhong Agricultural University, Key Laboratory of Freshwater Biodiversity Conservation and Utilization, Ministry of Agriculture, Wuhan, People's Republic of China
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16
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Fujiki H, Suganuma M. Carcinogenic aspects of protein phosphatase 1 and 2A inhibitors. PROGRESS IN MOLECULAR AND SUBCELLULAR BIOLOGY 2009; 46:221-54. [PMID: 19184590 DOI: 10.1007/978-3-540-87895-7_8] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
Abstract
Okadaic acid is functionally a potent tumor promoter working through inhibition of protein phosphatases 1 and 2A (PP1 and PP2A), resulting in sustained phosphorylation of proteins in cells. The mechanism of tumor promotion with okadaic acid is thus completely different from that of the classic tumor promoter phorbol ester. Other potent inhibitors of PP1 and PP2A - such as dinophysistoxin-1, calyculins A-H, microcystin-LR and its derivatives, and nodularin - were isolated from marine organisms, and their structural features including the crystal structure of the PP1-inhibitor complex, tumor promoting activities, and biochemical and biological effects, are here reviewed. The compounds induced tumor promoting activity in three different organs, including mouse skin, rat glandular stomach and rat liver, initiated with three different carcinogens. The results indicate that inhibition of PP1 and PP2A is a general mechanism of tumor promotion applicable to various organs. This study supports the concept of endogenous tumor promoters in human cancer development.
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Affiliation(s)
- Hirota Fujiki
- Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho, Tokushima 770-8514, Japan.
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17
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Seidler U, Lenzen H, Cinar A, Tessema T, Bleich A, Riederer B. Molecular mechanisms of disturbed electrolyte transport in intestinal inflammation. Ann N Y Acad Sci 2006; 1072:262-75. [PMID: 17057206 DOI: 10.1196/annals.1326.024] [Citation(s) in RCA: 49] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022]
Abstract
Diarrhea is the hallmark of both ulcerative colitis (UC) and Crohn's disease. Loss of resorptive area, destruction of epithelial cells, leaky tight junctions, and release of inflammatory mediators and products from immune cells that stimulate fluid secretion all have been implicated in the pathogenesis of inflammatory diarrhea. Very early studies in patients, however, have pinpointed the overwhelming transport abnormality in inflamed intestinal mucosa: a virtually complete loss of sodium resorptive capacity. Recently, tools have become available to study the molecular basis of disturbances in the major electrolyte transport systems during intestinal inflammation. This review gives a brief overview of the historical development of research related to electrolyte transport in inflammatory bowel disorders, focusing on the studies performed in humans, and highlights recent understanding of the molecular mechanisms that may help explain the origin of diarrhea in intestinal inflammation.
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Affiliation(s)
- Ursula Seidler
- Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Hannover, Germany.
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Goral J, Kovacs EJ. In vivo ethanol exposure down-regulates TLR2-, TLR4-, and TLR9-mediated macrophage inflammatory response by limiting p38 and ERK1/2 activation. THE JOURNAL OF IMMUNOLOGY 2005; 174:456-63. [PMID: 15611271 DOI: 10.4049/jimmunol.174.1.456] [Citation(s) in RCA: 105] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
Ethanol is known to increase susceptibility to infections, in part, by suppressing macrophage function. Through TLRs, macrophages recognize pathogens and initiate inflammatory responses. In this study, we investigated the effect of acute ethanol exposure on murine macrophage activation mediated via TLR2, TLR4, and TLR9. Specifically, the study focused on the proinflammatory cytokines IL-6 and TNF-alpha and activation of p38 and ERK1/2 MAPKs after a single in vivo exposure to physiologically relevant level of ethanol followed by ex vivo stimulation with specific TLR ligands. Acute ethanol treatment inhibited IL-6 and TNF-alpha synthesis and impaired p38 and ERK1/2 activation induced by TLR2, TLR4, and TLR9 ligands. We also addressed the question of whether ethanol treatment modified activities of serine/threonine-specific, tyrosine-specific phosphatases, and MAPK phosphatase type 1. Inhibitors of three families of protein phosphatases did not restore ethanol-impaired proinflammatory cytokine production nor p38 and ERK1/2 activation. However, inhibitors of serine/threonine protein phosphatase type 1 and type 2A significantly increased IL-6 and TNF-alpha levels, and prolonged activation of p38 and ERK1/2 when triggered by TLR4 and TLR9 ligands. In contrast, with TLR2 ligand stimulation, TNF-alpha production was reduced, whereas IL-6 levels, and p38 and ERK1/2 activation were not affected. In conclusion, acute ethanol exposure impaired macrophage responsiveness to multiple TLR agonists by inhibiting IL-6 and TNF-alpha production. Mechanism responsible for ethanol-induced suppression involved inhibition of p38 and ERK1/2 activation. Furthermore, different TLR ligands stimulated IL-6 and TNF-alpha production via signaling pathways, which showed unique characteristics.
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Affiliation(s)
- Joanna Goral
- Department of Cell Biology, Neurobiology and Anatomy, Loyola University Medical Center, 2160 South First Avenue, Maywood, IL 20153, USA
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Nobre ACL, Nunes-Monteiro SM, Monteiro MCSA, Martins AMC, Havt A, Barbosa PSF, Lima AAM, Monteiro HSA. Microcystin-LR promote intestinal secretion of water and electrolytes in rats. Toxicon 2004; 44:555-9. [PMID: 15450931 DOI: 10.1016/j.toxicon.2004.07.014] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2004] [Accepted: 07/12/2004] [Indexed: 01/01/2023]
Abstract
We showed previously that exposure to microcystin-LR causes renal toxic effects in isolated perfused rat kidney, and that inflammatory mediators from supernatants of macrophages stimulated by microcystin-LR are involved in this process. The aim of this research was to examine water and electrolytes secretion in vivo, induced by microcystin-LR and supernatant of macrophages stimulated for this toxin (SUP.MphiS + MCLR), using perfused rat ileal segment and ligated intestinal loop models. We found microcystin-LR at 1 microg/ml (0.09 +/- 0.003* vs. control 0.07 +/- 0.001 g of secretion/2 cm of loop; P < 0.05*) and the SUP.MphiS + MCLR after 18 h postinoculation (0.10 +/- 0.003 vs. control 0.03 +/- 0.002 g/cm) caused intestinal secretion. In addition, microcystin-LR caused significant sodium secretion (-2.18 +/- 0.72* vs. control 2.18 +/- 0.50 microEq g(-1) min(-1)), potassium (-0.26 +/- 0.04* vs. control 0.32 +/- 0.03 microEq g(-1) min(-1)), chloride (MCLR = -3.29 +/- 1.93* vs. control 0.88 +/- 1.25 microEq g(-1) min(-1)) and water (-0.012 +/- 0.004* vs. control 0.002 +/- 0.002 ml g(-1) min(-1)). We also demonstrated SUP.MphiS + MCLR to induce intestinal secretion of electrolytes (sodium, potassium, chloride) and water. These findings suggested that microcystin-LR and lamina propria macrophages-derived mediators are able to induce intestinal secretion in vivo, probably via inhibition of protein phosphatase.
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Affiliation(s)
- A C L Nobre
- Department of Clinical and toxicological Analysis, Federal University of Ceará, Fortaleza, Brazil
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Shi Q, Cui J, Zhang J, Kong FX, Hua ZC, Shen PP. Expression modulation of multiple cytokines in vivo by cyanobacteria blooms extract from taihu lake, China. Toxicon 2004; 44:871-9. [PMID: 15530969 DOI: 10.1016/j.toxicon.2004.08.010] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/02/2004] [Accepted: 08/19/2004] [Indexed: 12/19/2022]
Abstract
Cyanobacterial blooms that generate microcystins (MCs) are being increasingly recognized as a potent health hazard in aquatic ecosystems. However, immunomodulation induced by cyanotoxins has not been well documented. This paper reports the in vivo data on the immune disorder caused by crude microcystin (MC) extract of cyanobacteria blooms collected from Taihu Lake, China, with respect to cytokine mRNA levels. Using reverse-transcriptional polymerase chain reaction (RT-PCR), the expression of multiple cytokines, including proinflammatory (IL-1beta, TNF-alpha, and IL-6) and Th1/Th2-related cytokines (IL-2, IL-4 and IL-10), was evaluated following the cyanobacteria blooms extract containing MCs (CBE) exposure at four doses of 23, 38, 77, 115 mg lyophilized algae cells/kg body weight. The results showed that the mRNA levels of TNF-alpha, IL-1beta, IL-2 and IL-4 decreased significantly following injection of all doses as compared to the control (LPS or ConA only), while the IL-6 level was unaffected. Contrast to this decrease, the level of IL-10 mRNA was, however, transiently up regulated following injection of the lowest dose of CBE. The distinct patterns of expression of these cytokines suggested a modulation of cytokine network, the essential component of the host immune system. We further developed a mathematical model to simulate the interaction of T helper cell subsets and related cytokines, which proved to be a good approach to study the kinetics of the interaction of cells and cytokines in microcystin immunosuppression.
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Affiliation(s)
- Q Shi
- Department of Biochemistry, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China
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Lankoff A, Carmichael WW, Grasman KA, Yuan M. The uptake kinetics and immunotoxic effects of microcystin-LR in human and chicken peripheral blood lymphocytes in vitro. Toxicology 2004; 204:23-40. [PMID: 15369846 DOI: 10.1016/j.tox.2004.05.016] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2004] [Accepted: 05/13/2004] [Indexed: 10/26/2022]
Abstract
Microcystin-LR is a cyanobacterial heptapeptide that presents acute and chronic hazards to animal and human health. We investigated the influence of this toxin on human and chicken immune system modulation in vitro. Peripheral blood lymphocytes were treated with microcystin-LR at environmentally relevant doses of 1, 10 and 25 microg/ml for 12, 24, 48, 72 h (for proliferation assay cells were treated for 72 h). T-cell and B-cell proliferation as well as apoptosis and necrosis were determined in human and chicken samples. IL-2 and IL-6 production by human lymphocytes also was measured. In addition, uptake kinetics of microcystin-LR into human and chicken peripheral blood lymphocytes were calculated by Liquid Chromatography (LS) /Mass Spectrometry (MS) analysis. At the highest dose microcystin-LR decreased T-cell proliferation and all doses of microcystin-LR inhibited B-cell proliferation. The frequency of apoptotic and necrotic cells increased in a dose and time-dependent manner. Human lymphocytes responded to stimulation with microcystin-LR by increased production of IL-6 and decreased production of IL-2. Human lymphocytes were able to uptake from 0.014 to 1.663 microg/ml and chicken lymphocytes from 0.035 to 1.733 microg/ml of the microcystin-LR added to the cultures, depending on the treatment time and dose. In conclusion, microcystin-LR acted as an immunomodulator in cytokine production and down-regulated lymphocyte functions by induction of apoptosis and necrosis. However, further studies dealing with the influence of microcystin-LR on expression cytokine genes and transcription factors are necessary to confirm these hypotheses.
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Affiliation(s)
- Anna Lankoff
- Department of Radiobiology and Immunology, Institute of Biology, Pedagogical University, Swietokrzyska Academy, 25-406 Kielce, ul. Swietokrzyska, 1525 425, Poland.
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Chen T, Zhao X, Liu Y, Shi Q, Hua Z, Shen P. Analysis of immunomodulating nitric oxide, iNOS and cytokines mRNA in mouse macrophages induced by microcystin-LR. Toxicology 2004; 197:67-77. [PMID: 15003335 DOI: 10.1016/j.tox.2003.12.013] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2003] [Accepted: 12/27/2003] [Indexed: 10/26/2022]
Abstract
Microcystins (MCs) are the toxic molecules produced by common cyanobacterium in freshwater blooms. Their toxicities have brought severe health issues to livestock and human being. Microcystin-LR (MC-LR) is one of the most toxic MCs. This paper presents the profile of the immunomodulation of MC-LR to BALB/c mice peritoneal macrophages. Macrophages were stimulated with 100microg/l lipopolysaccharide (LPS) and MC-LR at dose of 1, 10, 100, 1000nmol/l, respectively, for 24h. Nitric oxide (NO) production in cell culture supernatants was quantified by using Griess reagent method. Total RNA was extracted from incubated macrophages then the mRNA abundance of induced nitric oxide synthase (iNOS), IL-1beta, TNF-alpha, GM-CSF, IFN-gamma was monitored by using reverse-transcriptional polymerase chain reaction (RT-PCR). The results demonstrated that NO production, mRNA levels of iNOS, IL-1beta, TNF-alpha were down regulated by MC-LR dose-dependently and mRNA levels of GM-CSF and IFN-gamma were also decreased but in dose-independent manner. Our results illustrated the involvement of NO production, iNOS and some cytokines in mice immune system in microcystin shock.
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MESH Headings
- Adjuvants, Immunologic/toxicity
- Animals
- Cell Survival/drug effects
- Cells, Cultured
- Culture Media, Conditioned/chemistry
- Cytokines/genetics
- Cytokines/metabolism
- Dose-Response Relationship, Drug
- Down-Regulation/drug effects
- Lipopolysaccharides/pharmacology
- Macrophages, Peritoneal/drug effects
- Macrophages, Peritoneal/metabolism
- Macrophages, Peritoneal/pathology
- Male
- Marine Toxins/toxicity
- Mice
- Mice, Inbred BALB C
- Microcystins
- Mitogens/pharmacology
- Nitric Oxide/metabolism
- Nitric Oxide Synthase/genetics
- Nitric Oxide Synthase/metabolism
- Nitric Oxide Synthase Type II
- Peptides, Cyclic/toxicity
- RNA, Messenger/metabolism
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Affiliation(s)
- Ting Chen
- Department of Biochemistry, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China
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Rocha MFG, Aguiar JEP, Sidrim JJC, Costa RB, Feitosa RFG, Ribeiro RA, Lima AAM. Role of mast cells and pro-inflammatory mediators on the intestinal secretion induced by cholera toxin. Toxicon 2003; 42:183-9. [PMID: 12906889 DOI: 10.1016/s0041-0101(03)00131-4] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Recent data suggest that diarrhea caused by Vibrio cholerae involves a pro-inflammatory mediators release, such as cytokines, prostaglandin and nitric oxide. The aim of this study was to investigate the role of mast cells and their mediators in the intestinal secretion induced by cholera toxin. We examined the dose responses, time course and role of mast cells and pro-inflammatory mediators in cholera toxin intestinal secretory response, in vivo. Cholera toxin caused a dose-dependent secretion, in ligated small intestine loops, at 18 h. Rats treated with 48/80 compound or ketotifen had a significant decrease in the intestinal secretory response. Cholera toxin secretion was significantly reduced by an unspecific histamine/serotonin receptor antagonist, histamine receptor antagonist, phospholipase A2 and cyclooxygenase inhibitors, platelet-activating factor (PAF) receptor antagonists and TNF-alpha synthesis blockers. On the other hand, pretreatment with a specific serotonin receptor antagonist and lipoxygenase inhibitors failed to block this effect. Analysis of the intestinal fluid from rats injected with cholera toxin, revealed that cholera toxin induces the release of IL-1beta and TNF-alpha into fluid. The data suggest that, at least in part, mast cells are involved in cholera toxin-induced secretion, as well as point to the importance of histamine, prostaglandins, PAF, IL-1beta and TNF-alpha in this process.
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Affiliation(s)
- Marcos F G Rocha
- Faculty of Veterinary, State University of Ceará, Fortaleza, CE, Brazil.
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Nobre ACL, Martins AMC, Havt A, Benevides C, Lima AAM, Fonteles MC, Monteiro HSA. Renal effects of supernatant from rat peritoneal macrophages activated by microcystin-LR: role protein mediators. Toxicon 2003; 41:377-81. [PMID: 12565761 DOI: 10.1016/s0041-0101(02)00334-3] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
We have demonstrated previously that microcystin-LR promoted some renal alterations using the isolated perfused rat kidney preparation. However, these effects were not proved to be direct or indirect. The aim of the current work is to examine the renal effects promoted by supernatants from rat macrophages stimulated with microcystin-LR and the role of inflammatory mediators. Peritoneal macrophages were collected previously and were incubated for 1h in fresh medium (control) and in medium containing microcystin-LR. Dexamethasone, quinacrine, thalidomide and cycloheximide were administered 30 min before microcystin-LR. Supernatants of macrophages stimulated with or without pharmacological inhibitors were added on the perfused rat kidney model. The infusion of macrophages supernatants stimulated by microcystin-LR caused significant increases in renal vascular resistance (C: 4.93+/-0.33 vs T: 5.15+/-0.21), glomerular filtration rate (C: 0.559+/-0.008 vs T: 0.978+/-0.15) and urinary flow (C: 0.16+/-0.01 vs T: 0.23+/-0.03). Cycloheximide, quinacrine and dexamethasone blocked these effects and thalidomide blocked renal vascular resistance. Macrophages stimulated by microcystin-LR release mediators capable of promoting nephotoxicity in isolated perfused rat kidney. Phospholipase A(2), TNF-alpha and other protein mediators appear to be involved on its renal toxic mechanism.
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Affiliation(s)
- A C L Nobre
- Department of Physiology and Pharmacology, Faculty of Medicine, Federal University of Ceara, Rua Cel Nunes de Melo, 1127, Fortaleza, CE 60430-270, Brazil
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