Liver diseases are a significant global health issue, characterized by elevated levels of disorder and death. The substantial impact of ageing on liver diseases and their prognosis is evident. Multiple processes are involved in the ageing process, which ultimately leads to functional deterioration of this organ. The process of liver ageing not only renders the liver more susceptible to diseases but also compromises the integrity of other organs due to the liver's critical function in metabolism regulation. A growing body of research suggests that long non-coding RNAs (lncRNAs) play a significant role in the majority of pathophysiological pathways. They regulate gene expression through a variety of interactions with microRNAs (miRNAs), messenger RNAs (mRNAs), DNA, or proteins. LncRNAs exert a major influence on the progression of age-related liver diseases through the regulation of cell proliferation, necrosis, apoptosis, senescence, and metabolic reprogramming. A concise overview of the current understanding of lncRNAs and their potential impact on the development of age-related liver diseases will be provided in this mini-review.

While Hepatitis A Virus (HAV) vaccination in global immunization programs has shown a virtual elimination of the disease within few years of the vaccination program, changing epidemiological landscape in India underscores the need for evidence-based, updated guidance on immunization practices. In May 2024, a panel of 15 distinguished opinion leaders and an organizing committee convened for an intensive, face-to-face advisory board meeting on high burden of HAV infection among adults, increased mortality rate in adolescents, symptomatic presentation in children, and evolving landscape globally and within India. Extensive comparable deliberations on long-term follow-up data from India and data from country of origin advocated immunogenicity, tolerability, and long-term protective effects of single-dose live attenuated HAV vaccine in children. Finally, a consensus was achieved on recognition of increased global attention toward HAV prevention through vaccination coverage. The need for a single dose of live attenuated HAV vaccine was an important outcome of this meeting.

Hepatitis B (Hep B) remains a critical public health issue globally, particularly in Tibet, where vaccination rates and influencing factors among college students are yet understudied. This study applies a cross-sectional design to investigate the Hep B vaccination rate among 1,126 college students in Tibet and utilizes the expanded theory of planned behavior (ETPB) to identify vaccination behavior intention (BI) and vaccination behavior (VB). Stratified cluster sampling across three universities was used to assess behavioral attitudes (BA), subjective norms (SN), perceived behavioral control (PBC), past vaccination history (PVH) and vaccination knowledge (VK), and used structural equation modeling (SEM) for model validation and multi-group comparison. Results indicated that 16.3% of students had received the Hep B vaccine. VK notably improved BA toward vaccination (β = 0.518, p < .001). BA (β = 0.232, p < .001), PBC (β = 0.239, p < .001), SN (β = 0.385, p < .001) positively influenced BI. However, PVH failed to predict BI. BI (β = 0.448, p < .001) and PVH (β = 0.127, p < .001) were significant predictors of VB. Significant ethnic variations were noted. The positive effect of PVH on VB (β = 0.151, p < .001) and the mediating role of PBC in VB (β = 0.076, p < .05) were significant among Tibetan students. The effect of VK on BA was stronger among Tibetans (β = 0.503, p < .05),while the impact of attitude on BI was more pronounced among Han students (β = 0.366, p < .05). The vaccination rate for Hep B among college students in Tibet is relatively low, and the ETPB model effectively explains their vaccination intentions and behaviors. Tailored intervention strategies for Tibetan and Han students are recommended to boost vaccination rates effectively.

Influenza causes 100,000-710,000 hospitalizations annually in the U.S. Patients with liver disease are at higher risk of severe outcomes following influenza infection. This study evaluated influenza vaccine effectiveness (VE) against influenza-associated hospitalization among adults with liver disease. Data from the U.S. Hospitalized Adult Influenza Vaccine Effectiveness Network (HAIVEN), a test-negative case-control study, from 2015 to 2020 were used to estimate VE among adults ≥18 years admitted for acute respiratory illness. VE was calculated as (1-adjusted odds ratio)*100%, comparing the odds of vaccine receipt between laboratory-confirmed influenza cases and test-negative controls using multiple logistic regression with inverse probability of treatment weighting (IPTW). In total, 1,622 (12.8%) of 12,704 adults had ≥1 liver disease(s). Compared with those without liver disease, adults with liver disease were more likely to be admitted to the intensive care unit (15.7% vs 12.8%, p = .001) or to die in hospital (3.0% vs 1.4%, p < .001). The IPTW-adjusted VE against influenza-associated hospitalization was 27% (95% confidence interval [CI], 22-32%) among patients without liver disease, but the VE of 11% (95% CI, -8-26%) was not significant among those with liver disease. Significant effect modification of VE by the presence of liver disease was found (p < .05 for interaction term). While influenza vaccination significantly reduced the risk of influenza-associated hospitalization among adults without liver disease, the protective effect was not significant among those with liver disease. Further studies are warranted to evaluate influenza VE in patients with different types of liver disease and with specific vaccine formulations.

This study aimed to investigate the potential causal association between chronic hepatitis B (CHB) infection and the risk of pre-eclampsia/eclampsia using a Mendelian randomisation (MR) design.

We conducted a two-sample MR analysis using genome-wide association study (GWAS) summary statistics from three large-scale datasets. For CHB infection, we used data from 351,885 individuals UK Biobank. For pre‑eclampsia/eclampsia, we analysed two FinnGen datasets with sample sizes of 118,291 and 126,760 individuals, respectively. Genetic variants strongly associated with CHB infection (p < 5 × 10-8) were selected as instrumental variables. The inverse-variance weighted (IVW) method was employed as the primary analysis. Sensitivity analyses included MR-Egger regression, weighted median, weighted mode and MR-PRESSO. Cochran's Q test and MR-Egger intercept tests were performed to assess heterogeneity and horizontal pleiotropy, respectively.

MR analysis revealed significant positive genetic associations between CHB infection and increased risk of pre-eclampsia (OR = 1.154, 95%CI = 1.014-1.313, p = .029) and eclampsia (OR = 1.561, 95%CI = 1.030-2.366, p = .035). Findings were robust across sensitivity analyses for both outcomes.

Our study provides genetic evidence that CHB infection increases the risk of both pre-eclampsia and eclampsia. These findings suggest that considering CHB status as a risk factor and implementing targeted HBV screening programmes may be beneficial for pregnant women.

Liver disease, responsible for around two million deaths annually, remains a pressing global health challenge. Microbial interactions within the microbiota-gut-liver axis play a substantial role in the pathogenesis of various liver conditions, including early chronic liver disease (eCLD), chronic liver disease (CLD), acute liver failure (ALF), acute-on-chronic liver failure (ACLF), non-alcoholic fatty liver disease (NAFLD), steatohepatitis, and cirrhosis. This study aimed to identify key molecular signatures involved in liver disease progression by analyzing transcriptomic and gut microbiome data, and to evaluate their diagnostic utility using machine learning models.

Transcriptomic analysis identified differentially expressed genes (DEGs) that, when integrated with regulatory elements microRNAs, transcription factors, receptors, and the gut microbiome highlight disease-specific molecular interactions. To assess the diagnostic potential of these molecular signatures, a two-step analysis involving principal component analysis (PCA) and Random Forest classification was conducted, achieving accuracies of 75% for ALF and 89% for NAFLD. Additionally, machine learning algorithms, including K-neighbors, multi-layer perceptron (MLP), decision tree, Random Forest, logistic regression, gradient boosting, CatBoost, Extreme Gradient Boosting (XGB), and Light Gradient Boosting Machine (LGBM), were applied to gene expression data for ALF and NAFLD.

Key genes including CLDN14, EGFR, GSK3B, MYC, and TJP2, alongside regulatory miRNAs let-7a-5p, miR-124-3p, and miR-195-5p and transcription factors NFKB1 and SP1 may be suggested as critical to liver disease progression. Additionally, gut microbiota members, Dictyostelium discoideum and Eikenella might be novel candidates associated with liver disease, highlighting the importance of the gut-liver axis. The Random Forest model reached 75% accuracy and 83% area under the curve for ALF, while NAFLD classification achieved 100% accuracy, precision, recall, and area under the curve underscoring robust diagnostic potential.

This study establishes a solid foundation for further research and therapeutic advancement by identifying key biomolecules and pathways critical to liver disease. Additional experimental validation is needed to confirm clinical applicability.

The dual barriers formed by the capillarized liver sinusoids and excessive deposited extracellular matrix (ECM) significantly impede the retention of therapeutic agents in the fibrotic liver. Currently, there are limited reports on strategies capable of simultaneously overcoming these barriers. Here, we propose sulfur dioxide (SO2)-releasing nanomotors based on endogenous in vivo reactions to restore the fenestrae of sinusoids and degrade ECM by activating the endogenous signaling pathway to improve the retention of therapeutic agents in the damaged liver. These nanomotors leverage the specific enzyme concentration gradient in damaged liver tissue as a chemoattractant signal, guiding their targeted delivery. The nanomotors incorporate an l-cysteine-based substrate that, upon enzymatic catalysis, generates SO2. The released SO2 can upregulate the cyclic guanosine monophosphate expression to restore the fenestrated phenotype of liver sinusoidal endothelial cells. Concurrently, SO2 can stimulate the endogenous nitric oxide production to induce matrix metalloproteinase-1 activation to facilitate the collagen degradation. The animal experimental model also demonstrates the effective retention of nanomotors in damaged liver tissue and reversal of liver fibrosis.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality, characterized by a complex molecular landscape. EGF Domain Specific O-linked β-N-acetylglucosamine transferase (EOGT) functions as an O-GlcNAc transferase with specific activity towards proteins containing epidermal growth factor (EGF) repeats. Although it is known to potentially play an oncogenic role in HCC, the exact mechanisms remain unclear. Induction of ferroptosis is a primary mechanism by which anticancer drugs such as sorafenib treat HCC. This study aimed to elucidate the expression profile of EOGT in HCC and its relationship with ferroptosis, as well as to investigate the underlying molecular mechanisms.

Utilizing bioinformatics resources, we explored the potential role of EOGT in HCC. The effects of EOGT on HCC cell behavior were examined using cell models and subcutaneous xenograft models in nude mice. Further insights into the molecular mechanisms were obtained through RNA-seq in cell models, hydrodynamic modeling in mice, Western blotting, chromatin immunoprecipitation (ChIP) sequencing, and dual-luciferase reporter assays to analyze the interaction between HEY1 and SLC7A11. Multiple validation steps were employed to thoroughly investigate the roles of these factors in the regulation of ferroptosis in HCC.

Our findings revealed that EOGT is upregulated in HCC and correlates with poor prognosis and drug resistance. Knockdown of EOGT inhibited HCC cell proliferation and enhanced sensitivity to ferroptosis by downregulating SLC7A11, a process mediated by HEY1. These results were confirmed by cell viability assays, quantitative real-time PCR (qPCR), Western blotting, and dual-luciferase reporter gene assays.

EOGT promotes HCC proliferation and inhibits ferroptosis by modulating the HEY1-SLC7A11 axis, suggesting a potential therapeutic target for HCC treatment.

This investigation explored the therapeutic effects and mechanisms of Skullcapflavone II in hepatic fibrosis (HF).

The optimal concentration of Skullcapflavone II for LX2 hepatic stellate cells was determined using the CCK8 assay. EdU staining and flow cytometry were utilised to assess cell proliferation and G2/M phase arrest. Mice with carbon tetrachloride-triggered HF were administered Skullcapflavone II at low (15 mg/day), medium (30 mg/day), and high (60 mg/day) doses. Subsequently, hepatic damage and fibrosis were assessed via body weight, liver index, biochemical markers, and histopathological staining. Immunohistochemistry for Collagen I and α-SMA were utilised to examine hepatic stellate cell (HSC) activation. RNA sequencing was utilised to ascertain differentially expressed genes. Molecular docking simulated interactions among Skullcapflavone II and target proteins as well as outcomes were validated by implementing western blotting, immunohistochemistry, and RT-qPCR.

Skullcapflavone II inhibited LX2 cell proliferation and triggered G2/M phase arrest. Its optimal intervention concentration was 160 μM. In vivo, it ameliorated hepatic function, diminished serum indicators of fibrosis, and suppressed HSC activation. Diminished collagen sediment was validated utilising histopathological examination, whereas immunohistochemistry indicated decreased expression of Collagen I and α-SMA. Additionally, molecular docking showed strong binding of Skullcapflavone II to DNA replication-related proteins. Western blotting and RT-qPCR implied that Skullcapflavone II disrupted DNA replication, which triggered G2/M arrest and hindered HSCs activation and proliferation.

The abovementioned mechanisms of action of Skullcapflavone II substantiate its prospective clinical application against HF.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major cause of chronic liver disease, primarily due to insulin resistance and type 2 diabetes (T2D). Despite the strong link between T2D and MASLD, identifying and treating liver fibrosis in T2D patients is still poor. This study aimed to identify the factors related to advanced liver fibrosis in T2D patients.

This retrospective observational study used medical records from four centers in Mexico City from 2018 to 2023. The study included 2000 patients with T2D. Liver fibrosis was evaluated using the Fibrosis-4 (FIB-4) index, and insulin resistance was assessed using the estimated glucose disposal rate (eGDR).

The mean age of the patients was 58.9 years, with 63.7% being women. The median duration of T2D was 7 years, and the mean HbA1c was 7.63%. Overall, 20.4% had advanced liver fibrosis. The multivariate logistic regression analysis showed that diabetes duration >10 years {odds ratio (OR) = 2.105 (95% confidence interval [CI] 1.321-3.355)}, fasting glucose >126 mg/dL (OR = 1.568 [95% CI 1.085-2.265]), and microalbuminuria >300 mg/24 h (OR = 2.007 [95% CI 1.134-3.552]) were associated with advanced liver fibrosis. Conversely, the eGDR (OR = 0.805 [95% CI 0.703-0.888]), statins (OR = 0.111 [95% CI 0.073-0.168]), and pioglitazone (OR = 0.082 [95% CI 0.010-0.672]) were inversely associated.

Longer diabetes duration, insulin resistance, and microalbuminuria are independently linked to advanced liver fibrosis in T2D patients. Statins and pioglitazone may protect against liver fibrosis. Enhanced screening and management strategies targeting these factors could slow fibrosis progression and reduce the global burden of MASLD.

Steatotic liver disease (SLD), which encompasses various causes of fat accumulation in the liver, is a major cause of liver fibrosis. Understanding the specific mechanisms of lipotoxicity, dysregulated lipid metabolism, and the role of different hepatic cell types involved in fibrogenesis is crucial for therapy development.

We analysed liver tissue from SLD patients and 3 mouse models. We combined bulk/spatial lipidomics, transcriptomics, imaging mass cytometry (IMC) and analysis of published spatial and single-cell RNA sequencing (scRNA-seq) data to explore the metabolic microenvironment in fibrosis. Pharmacological inhibition of sphingolipid metabolism with myriocin, fumonisin B1, miglustat and D-PDMP was carried out in hepatic stellate cells (HSCs) and human precision cut liver slices (hPCLSs).

Bulk lipidomics revealed increased glycosphingolipids, ether lipids and saturated phosphatidylcholines in fibrotic samples. Spatial lipidomics detected >40 lipid species enriched within fibrotic regions, notably sphingomyelin (SM) 34:1. Using bulk transcriptomics (mouse) and analysis of published spatial transcriptomics data (human) we found that sphingolipid metabolism was also dysregulated in fibrosis at transcriptome level, with increased gene expression for ceramide and glycosphingolipid synthesis. Analysis of human scRNA-seq data showed that sphingolipid-related genes were widely expressed in non-parenchymal cells. By integrating spatial lipidomics with IMC of hepatic cell markers, we found excellent spatial correlation between sphingolipids, such as SM(34:1), and myofibroblasts. Inhibiting sphingolipid metabolism resulted in anti-fibrotic effects in HSCs and hPCLSs.

Our spatial multi-omics approach suggests cell type-specific mechanisms of fibrogenesis involving sphingolipid metabolism. Importantly, sphingolipid metabolic pathways are modifiable targets, which may have potential as an anti-fibrotic therapeutic strategy.

The progression from hepatitis to liver fibrosis (LF) and ultimately to hepatic carcinoma (HCC) represents the advanced stages of various liver diseases. Currently, no universal treatment effectively addresses all three conditions. The Traditional Chinese Medicine formula Xiao-Chai-Hu decoction (XCHD) has shown promise in treating hepatitis, inhibiting LF, and serving as an adjunct therapy for HCC. This study evaluates the efficacy and optimal treatment durations of XCHD in managing these liver diseases using meta-analysis and machine learning techniques.

Registered in the PROSPERO database (CRD42024534445), this meta-analysis systematically searched seven databases, including 54 studies with a total of 5,710 patients. Statistical analysis was performed using Stata 17.0. Five machine learning models-Random Forest (RF), XGBoost, Lasso, Multilayer Perceptron (MLP), and a stacking model combining these algorithms-were employed to analyze the data and predict the time-effect relationships. The optimal durations of XCHD treatment for the liver disease trilogy were subsequently projected.

XCHD significantly improved the primary outcome indicators for hepatitis, liver fibrosis, and HCC. Additionally, XCHD demonstrated a beneficial effect on liver dysfunction caused by these diseases. Machine learning predicted the optimal treatment durations of XCHD as 12 weeks for hepatitis, 20.31 weeks for liver fibrosis, and 12 weeks for HCC.

XCHD is effective in treating the liver disease trilogy, with optimal treatment durations of 12 weeks for hepatitis and HCC, and 20.31 weeks for liver fibrosis. These findings support the potential of XCHD in developing precise clinical strategies for managing liver diseases. This study innovatively integrates meta-analysis with machine learning to determine the optimal treatment durations, providing a novel approach for evidence-based precision medicine in Traditional Chinese Medicine.

Non-alcoholic fatty liver disease (NAFLD) patients have multiple metabolic disturbances, with markedly elevated levels of lactate. Lactate accumulations play pleiotropic roles in disease progression through metabolic rearrangements and epigenetic modifications. Monocarboxylate transporter 4 (MCT4) is highly expressed in hepatocytes and responsible for transporting intracellular lactate out of the cell. To explore whether elevated MCT4 levels played any role in NAFLD development, we overexpressed and silenced MCT4 in hepatocytes and performed a comprehensive in vitro and in vivo analysis. Our results revealed that MCT4 overexpression down-regulated the genes for lipid synthesis while up-regulating the genes involved in lipid catabolism. Conversely, silencing MCT4 expression or inhibiting MCT4 expression led to the accumulation of intracellular lipid and glucose metabolites, resulting in hepatic steatosis. In a mouse model of NAFLD, we found that exogenous MCT4 overexpression significantly reduced lipid metabolism and alleviated hepatocellular steatosis. Mechanistically, MCT4 alleviated hepatic steatosis by regulating a group of hub genes such as Arg2, Olr1, Cd74, Mmp8, Irf7, Spp1, and Apoe, which in turn impacted multiple pathways involved in lipid metabolism and inflammatory response, such as PPAR, HIF-1, TNF, IL-17, PI3K-AKT, Wnt, and JAK-STAT. Collectively, our results strongly suggest that MCT4 may play an important role in regulating lipid metabolism and inflammation and thus serve as a potential therapeutic target for NAFLD.

Alcoholic liver injury (ALI) represents a major international public health concern with no targeted pharmacological intervention and a dearth of clinical trial-approved medications for its management. The dried leaves of Schisandra chinensis (SCL) are rich in flavonoids, lignans, polysaccharides, and other active ingredients with anti-inflammatory, antioxidant, and antitumour activities and are commonly applied in the treatment of osteoarthritis, diabetes, and neurodegenerative diseases. The crude lignans of SCL have been reported to treat CCl4-induced acute liver injury, and SCL tea has also been reported to have hepatoprotective effects. The components of SCL are currently the focus of investigation; however, conclusive pharmacological studies on SCL in the treatment of ethanol-induced ALI are rare.

This study aimed to identify the bioactive components and elucidate the mechanism of action of SCL against ALI.

The optimal month for harvesting SCL was first determined using a cellular ALI model. Immediately afterwards, the efficacy of SCL was evaluated based on cellular ALI model and ALI mice model. The expression levels of NLRP3 inflammasome-associated proteins were examined via Western blotting. To identify the bioactive components of SCL, the common components in 10 batches of SCL were identified by UPLC‒Q-TOF‒MS/MS. Subsequent analysis via correlation identified common elements' pharmacological impacts, filtering for substances with notable contributions to effectiveness. Finally, potential bioactive components were further identified through molecular docking and verified in ALIcell models.

SCL has the best efficacy in early August, and by improving hepatic aminotransferase activity, regulating lipid metabolism, alleviating oxidative stress, reducing the release of inflammatory mediators, and inhibiting the expression of NLRP3-related pyroptosis proteins, it plays a role in alleviating ALI. A total of 32 common components were identified in 10 batches of SCL. Through correlation analysis, 10 functional components, including Schisandrin B, Angeloylgomisin Q, Chlorogenic acid, Rutin, Schisandrin C, p-Hydroxycinnamic acid, Schisandrin A, Schisandrol A, Gomisin J, and Schisantherin B, were screened. Further screening using molecular docking identified 4 key functional components, Rutin, Chlorogenic acid, Schisandrin C, and Schisantherin B, which were verified to mitigate ethanol-induced liver damage.

The present study demonstrated that SCL prevented ALI, with the main contributing components being rutin, chlorogenic acid, pentosidine C and pentosidine B. Hence, our latest study offers significant experimental proof indicating SCL as a promising prospect for ALI prevention.

Excessive alcohol consumption is a leading cause of alcohol-associated liver disease (ALD). Previous studies presented Mogroside V (MV) have protective effects on against nonalcoholic fatty liver disease. however, the effects of MV on ethanol-induced hangover and liver damage remains to be elucidated. Herein, we investigated the potential effects of MV in relieving hangover and mitigating liver injury induced by ethanol. MV significantly reduced blood ethanol, liver histological alterations and serum ALT, AST、TG levels in ethanol-treated mice. Moreover, MV accelerates alcohol metabolism by inhibiting the upregulation of CYP2E1 induced by ethanol, while enhancing the activity of ADH and ALDH, as well as upregulating the expression of ADH1 and ALDH2. MV mitigates oxidative stress by decreased hepatic malondialdehyde (MDA) levels, restored glutathione (GSH)、superoxide dismutase (SOD) and catalase (CAT) content in ethanol-induced mice. Mechanistically, MV activated the p-AMPK/SREBP-1/FASN pathway to decreased hepatic lipid accumulation and alleviated steatosis. Additionally, MV promoted nuclear translocation of Nrf-2 to attenuates oxidative stress and suppressed TLR4/MyD88/NF-κB signaling pathway to reduce Inflammatory responses triggered by ethanol in mice. In summary, this study highlights mogroside V's hangover relieving effect and its protective effects against ethanol-related liver damage through its lipid metabolism regulation, antioxidative action and anti-inflammatory properties. These results suggest that mogroside V could be developed as a potential therapeutic agent against ethanol-induced liver damage.

Polycyclic aromatic hydrocarbons (PAHs) exposure has been suggested to be linked to abnormal liver function (ALF). However, the conclusions are inconsistent, and the underlying mechanism is still unclear. In this study, a cross-sectional design including 4935 adults from the National Health and Nutrition Examination Survey (NHANES) between 2003 and 2010 was conducted to quantify the PAHs-ALF associations, and to investigate the possible mediation role of systemic inflammation. Capillary gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was utilized to detect nine urinary levels of PAH metabolites (OH-PAHs). Plasma levels of systemic inflammation biomarker, C-reactive protein (CRP), were measured by enhanced turbidimetry. ALF was diagnosed on the basis of any abnormities in albumin (ALB), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), and alanine aminotransferase (ALT). Logistic regression model and the least absolute shrinkage and selection operator (LASSO) regression models indicated positive associations between urinary 1-hydroxypyrene (1-OH-PYR), 2-hydroxyphenanthrene (2-OH-PHE), and ALF risk. Significant synergistic effect of 1-OH-PYR and 2-OH-PHE on ALF was observed via additive interaction analysis. The weighted quantile sum (WQS) analysis and the quantile-based g computation (qgcomp) were employed to investigate the mixed effect of PAHs but no significant results were found. However, these two analyses consistently showed that 2-OH-PHE and 1-OH-PYR had top dominant weights in the positive association with ALF. Furthermore, mediation analysis indicated that plasma CRP mediated 13.4 % of the association between 1-OH-PYR and ALF risk. These results enhanced our comprehension of the health effects of PAHs exposure on liver function as well as the underlying mechanism.

Nanocarriers can improve the therapeutic efficiency of small molecule immunomodulators or inhibitors, which is important for immunotherapy of liver diseases or cancer. Macromolecular protein carriers, such as human serum albumin (HSA), could provide better penetration compared to large nanoparticles (>50 nm) but are hampered by systemic biodistribution. To overcome these limitations, inspired by the natural glycogen structure, we have designed the HSA nanocarrier (<40 nm) consisting of multiple trimannose (TM) ligands attached to the protein surface, to target mannose or dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) receptors in immune cells or immunological organs such as the liver. Capitalizing on the chemical reactivity of different amino acids present in HSA, we have incorporated multiple copies of a cargo relevant for immunotherapy, i.e. the toll-like receptor (TLR) 7/8 agonist. The resulting TM-HSA conjugates exhibit excellent and specific uptake ex vivo in various immune cells and liver-specific uptake in vivo, opening access to protein nanocarriers with rapid and efficient in vivo targeting with great potential for immune-related diseases.

Various hepatocellular carcinoma (HCC) prediction models have been proposed for patients with chronic hepatitis B (CHB) using clinical variables. We aimed to develop an artificial intelligence (AI)-based HCC prediction model by incorporating imaging biomarkers derived from abdominal computed tomography (CT) images along with clinical variables.

An AI prediction model employing a gradient-boosting machine algorithm was developed utilizing imaging biomarkers extracted by DeepFore, a deep learning-based CT auto-segmentation software. The derivation cohort (n = 5,585) was randomly divided into the training and internal validation sets at a 3:1 ratio. The external validation cohort included 2,883 patients. Six imaging biomarkers (i.e. abdominal visceral fat-total fat volume ratio, total fat-trunk volume ratio, spleen volume, liver volume, liver-spleen Hounsfield unit ratio, and muscle Hounsfield unit) and eight clinical variables were selected as the main variables of our model, PLAN-B-DF.

In the internal validation set (median follow-up duration = 7.4 years), PLAN-B-DF demonstrated an excellent predictive performance with a c-index of 0.91 and good calibration function (p = 0.78 by the Hosmer-Lemeshow test). In the external validation cohort (median follow-up duration = 4.6 years), PLAN-B-DF showed a significantly better discrimination function compared to previous models, including PLAN-B, PAGE-B, modified PAGE-B, and CU-HCC (c-index, 0.89 vs. 0.65-0.78; all p <0.001), and maintained a good calibration function (p = 0.42 by the Hosmer-Lemeshow test). When patients were classified into four groups according to the risk probability calculated by PLAN-B-DF, the 10-year cumulative HCC incidence was 0.0%, 0.4%, 16.0%, and 46.2% in the minimal-, low-, intermediate-, and high-risk groups, respectively.

This AI prediction model, integrating deep learning-based auto-segmentation of CT images, offers improved performance in predicting HCC risk among patients with CHB compared to previous models.

The novel predictive model PLAN-B-DF, employing an automated computed tomography segmentation algorithm, significantly improves predictive accuracy and risk stratification for hepatocellular carcinoma in patients with chronic hepatitis B (CHB). Using a gradient-boosting algorithm and computed tomography metrics, such as visceral fat volume and myosteatosis, PLAN-B-DF outperforms previous models based solely on clinical and demographic data. This model not only shows a higher c-index compared to previous models, but also effectively classifies patients with CHB into different risk groups. This model uses machine learning to analyze the complex relationships among various risk factors contributing to hepatocellular carcinoma occurrence, thereby enabling more personalized surveillance for patients with CHB.

Liver damage caused by high doses of paracetamol is a global public health concern. Consequently, therapeutic strategies are being explored to prevent this damage. The bioactive compounds present in fruits have shown promise in protecting against disorders associated with paracetamol-induced liver damage. This study assessed the preventive effects of guarana powder on redox status in a rat model of acute hepatotoxicity induced by a toxic dose of paracetamol. Male Wistar rats were divided into four groups: control (C), guarana (G), paracetamol (P), and guarana + paracetamol (GP). Animals in groups G and GP received 300 mg/kg guarana powder daily for seven days. Hepatotoxicity was induced in the P and GP groups by a single dose of 3 g/kg paracetamol on the last day. Paracetamol effectively induced liver damage and oxidative stress in group P animals. Preventive treatment with guarana significantly mitigated this damage and prevented the serum elevation of ALT, AST, and ALP by 44 %, 29 %, and 24 %, respectively. It also prevented a 133 % increase in the necrotic liver area in GP animals compared to the P. Guarana treatment, which prevented reductions in glutathione levels, modulated antioxidant enzyme (SOD and CAT) expression and activity, and protein carbonylation, while enhancing the total antioxidant capacity. Our results suggest that preventive treatment with guarana can attenuate oxidative damage, modulate antioxidant defense gene expression, and protect against paracetamol-induced hepatotoxicity in rats, highlighting guarana powder as a potential therapeutic agent to prevent liver damage induced by high doses of paracetamol.

Patient-reported outcome measures (PROMs) are being used more often in chronic liver disease (CLD) clinical care and research. Their interpretability can be greatly enhanced by establishing the smallest meaningful score difference (MSD). We report scores of commonly used PROMs and their MSDs in patients at different stages of liver disease.

Patient-Reported Outcomes Measurement Information System (PROMIS)-29 Profile, Chronic Liver Disease Questionnaire (CLDQ), and Short Form-36 (SF-36) v1.0 scores were aggregated from 2442 adults with CLD at 4 different stages: inpatients with decompensated cirrhosis (n=1146) and outpatients with cirrhosis (n=677) or CLD (n=128) or recipients of liver transplant (LT, n=490) between June 2014 and April 2023 from 3 academic centers. MSDs were estimated using distribution and anchor-based methods.

The study sample's median age was 60.0 (IQR: 51.0-66.0); 55% were male, 17% Hispanic, 84% White, and 49% college educated. The etiology of CLD was alcohol in 36%, metabolic dysfunction-associated steatohepatitis (MASH) in 31%, and viral hepatitis B/C in 26%. Median PROMIS domain scores were generally lowest in inpatients and highest after transplant. For PROMIS, distribution-based and anchor-based MSDs ranged from 3 to 4 for individual domains and 4 to 6 for summary scores. Distribution-based MSDs were 1 for CLDQ and ranged from 7 to 11 for individual SF-36 domains, except role limitations domains, which ranged from 15 to 18, and component scores, which were 3. When compared across stages of liver disease, PROMIS MSDs were generally similar, although they tended to be 0.5-1.0 points smaller in the decompensated population compared to the stable populations.

This study provides data-driven recommendations for MSDs, enhancing the interpretability of commonly used PROMs in liver disease and facilitating the integration of PROMs in various clinical and research settings.

Alcoholic liver disease (ALD) is a major cause of chronic liver disease worldwide with no approved therapy. The development of ALD is strongly associated with hepatic lipid accumulation. Arrestin domain containing 3 (ARRDC3), a member of the α-arrestin family, is involved in obesity, inflammation, and cancer. However, its role in ALD remains largely unexplored.

Both the NIAAA and traditional Lieber-De Carli mouse models of ALD were employed. ARRDC3 expression was evaluated in liver specimens from ALD patients, mouse hepatic tissues, and hepatocytes. Hepatocyte-targeted Arrdc3 knockdown was achieved through intrahepatic delivery of adeno-associated virus 8 (AAV8) carrying shRNA under a hepatocyte-specific promoter. Mass spectrometry analysis, immunofluorescence, co-immunoprecipitation (co-IP) assays, and molecular docking were used to identify the interaction between ARRDC3 and stearoyl-CoA desaturase 1 (SCD1).

ARRDC3 levels were significantly elevated in the livers of both ALD patients and mouse models. Knockdown of Arrdc3 using AAV8 alleviated alcohol-induced liver steatosis in both the NIAAA and traditional Lieber-De Carli mouse models. We demonstrated that ARRDC3 promoted the progression of ALD by inducing lipid accumulation in hepatocytes. Mechanistically, ARRDC3 directly binds to SCD1 and inhibits its ubiquitin-proteasome degradation. Inhibition of SCD1 blocked ARRDC3-induced lipid deposition in hepatocytes. We also observed a correlation between ARRDC3 and SCD1 in liver samples from ALD patients.

Our findings reveal that ARRDC3 promotes hepatic steatosis in ALD by reducing the ubiquitin-dependent degradation of SCD1. ARRDC3 may serve as a potential therapeutic target for ALD.

Digital therapeutics have developed rapidly in recent years, providing a new method for disease management. The burden of liver diseases in China is heavy, and there are obvious problems in disease management. This paper expounds on the definition and classification of digital therapeutics, introduces their application in liver disease treatment and management in detail, and analyzes their development prospects in the field of liver diseases and future challenges.Graphical abstract available for this article.

Alcohol-related liver disease (ALD) is a major cause of mortality and disability-adjusted life years. It is not fully understood why a small proportion of patients develop progressive forms of ALD (e.g., fibrosis and cirrhosis). Differences in the metabolic processes could be behind the individual progression of ALD. Our aim was to examine differences in serum metabolome between patients with nonprogressive ALD and patients with an early form of progressive ALD.

The study had three study groups: progressive ALD (alcohol-related steatohepatitis or early-stage fibrosis, n = 50), nonprogressive ALD (simple steatosis, n = 50) and healthy controls (n = 32). Both ALD groups took part in a voluntary alcohol rehabilitation programme. A nontargeted metabolomics analysis and targeted analysis of short-chain fatty acids were done to the serum samples taken on the day of admission.

We found 111 significantly (p < 0.0005) altered identified metabolites between the study groups. Our main finding was that levels of glycine-conjugated bile acids (Cohen's d = 0.90-0.91), glutamic acid (d = 1.01), 7-methylguanine (d = 0.77) and several phosphatidylcholines (d = 0.61-0.85) were elevated in the progressive ALD group in comparison to the nonprogressive ALD group. Glycine-conjugated bile acids, glutamic acid and 7-methylguanine also positively correlated with increased levels of aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, cell death biomarker M65 and liver stiffness.

Our results indicate that the enterohepatic cycle of glycine-conjugated bile acids, as well as lipid and energy metabolism, is altered in early forms of progressive ALD. These metabolic processes could be a target for preventing the progression of ALD.

Cancer treatments such as surgery and chemotherapy have several limitations, including ineffectiveness against large or persistent tumors, high relapse rates, drug toxicity, and non-specificity of therapy. Researchers are exploring advanced strategies for treating this life-threatening disease to address these challenges. One promising approach is targeted drug delivery using prodrugs or surface modification with receptor-specific moieties for active or passive targeting. While various drug delivery systems have shown potential for reaching hepatic cells, nano-carriers offer significant size, distribution, and targetability advantages. Engineered nanocarriers can be customized to achieve effective and safe targeting of tumors by manipulating physical characteristics such as particle size or attaching receptor-specific ligands. This method is particularly advantageous in treating liver cancer by targeting specific hepatocyte receptors and enzymatic pathways for both passive and active therapeutic strategies. It highlights the epidemiology of liver cancer and provides an in-depth analysis of the various targeting approaches, including prodrugs, liposomes, magneto-liposomes, micelles, glycol-dendrimers, magnetic nanoparticles, chylomicron-based emulsion, and quantum dots surface modification with receptor-specific moieties. The insights from this review can be immensely significant for preclinical and clinical researchers working towards developing effective treatments for liver cancer. By utilizing these novel strategies, we can overcome the limitations of conventional therapies and offer better outcomes for liver cancer patients.

Hepatocyte nuclear factor 4 alpha (HNF4α) is a master regulator of hepatocyte differentiation in fetal and adult liver and exerts its transcriptional role in determining physiological functions of the liver. The objective of this review is to address the current knowledge of molecular mechanisms involved in HNF4α regulation in multiple aspects of liver disease pathogenesis.

Based on available literature, this review summarises the current state of knowledge onthe mechanism of HNF4α dysregulation, and the role of HNF4α activity inregulating early to advanced stages of various liver diseases.

Patients with deranged HNF4α expression are at higher risk for the development of liver diseases such as viral hepatitis, alcoholic/nonalcoholic fatty liver disease, hepatocellular carcinoma, and haematological disorders such as coagulopathy and bleeding disorders.

HNF4α interactions with nuclear receptors and target genes promote liver disease pathology by regulating various metabolic pathways. The strong correlation between deranged HNF4α expression and the severity of liver diseases suggests that targeting HNF4α expression can offer potential therapeutic strategy in the prevention of liver disease pathology and haemostatic disorders.

Alcohol-related cirrhosis (ALD cirrhosis) is a chronic liver disease caused by excessive alcohol consumption. Patients with ALD cirrhosis have increased risk of conditions that may require surgery for example gallstones, gastrointestinal cancers, and hernias, but surgeons may prefer to avoid surgery due to patients' increased risk of complications. We compared the incidence of surgery between patients with ALD cirrhosis and matched comparators.

We used nationwide healthcare registries to include all Danish citizens diagnosed with ALD cirrhosis in 1996-2018. Follow-up began at the time of cirrhosis diagnosis and ended at death or 10 years after diagnosis. We compared their incidence rates and cumulative risks of surgery (excluding biopsies, punctures, drainages, injections, and diagnostic endoscopies) with those for a gender- and age-matched comparator cohort from the general Danish population.

We included 23,050 patients with ALD cirrhosis and 113,412 matched comparators. The incidence rate ratio (IRR) for surgery overall was 3.38 [95% Confidence Interval (CI): 3.34-3.41] for patients vs. comparators. The 1-year risk of surgery was 42.6% [95% CI: 42.4-42.9] for patients vs. 13.4% [95% CI: 13.3-13.6] for comparators. Abdominal procedures were the most frequent surgical category for the patients with an IRR of 6.55 [95% CI: 6.41-6.69] for patients vs. comparators.

We found an increased risk of surgery for patients with ALD cirrhosis compared with the general Danish population. Given the frequency of surgery in these high-risk patients, further research should focus on perioperative management to optimize outcomes.

Globally, the aetiology and epidemiology of chronic liver disease (CLD) are undergoing significant changes. We aimed to investigate the updated global burden of CLD, evaluate the cross-country inequalities, and provide 2050 predictions.

Using the Global Burden of Disease Study (GBD) 2021 data resources, we analysed and forecasted CLD prevalence, incidence, and related death from 1990-2021 to 2050, respectively. We calculated average annual percent change (AAPC) by joinpoint regression model and quantified inequalities according to World Health Organisation-recommended health equity standards.

In 2021, the number of prevalent, incident CLD and related deaths globally were 1.7 billion (95% uncertainty interval (UI): 1.6-1.8), 58.4 million (95% UI: 54.2-62.8) and 1.4 million (95% UI: 1.3-1.5), respectively. During 1990-2021, the age-standardised incidence rate (ASIR) increased, especially in those aged 15-49 (AAPC: 0.49%; 95% confidence interval [CI]: 0.45%-0.53%), in Europe (AAPC: 0.41%; 95% CI: 0.41%-0.42%) and the Americas (AAPC: 0.41%; 95% CI: 0.39%-0.42%), whereas the age-standardised death rate (ASDR) decreased globally (AAPC: -1.26%; 95% CI: -1.35% [-1.17%]) and across subgroups. During 1990-2021, the ASIR of metabolic dysfunction-associated steatotic liver disease (MASLD) increased the most in those aged 15-49 (AAPC: 0.72%; 95% CI: 0.67%-0.77%) and in the Western Pacific region (AAPC: 0.73%; 95% CI: 0.59%-0.86%). Socio-demographic index (SDI)-related inequalities decreased for the age-standardised prevalence rate (ASPR) and ASIR of CLD but increased for ASDR, placing a disproportionately heavier burden on low-SDI countries. From 2022 to 2050, the ASIR of CLD is projected to increase (AAPC: 0.20%; 95% CI: 0.19%-0.20%), but the ASDR is projected to decline (AAPC: -1.91%; 95% CI: -1.96% [-1.85%]).

This study's findings highlight targeted interventions for CLD disparities, focusing on MASLD management, the younger population (15-49 years), and socio-demographic inequalities.

Liver injury and uveitis pose severe threats to human health. Owing to the close relationship of physiology and pathology between the liver and the eyes, cases in which both conditions occur simultaneously are not uncommon in clinical settings, significantly complicating treatment. However, no suitable comorbid animal model has been reported, and research on the pathological mechanisms of this comorbidity is lacking. Prunella vulgaris L., a well-known traditional Chinese medicine renowned for its liver-clearing and eye-brightening properties. Prunella vulgaris polyphenols (PVPs) hold promise for improving liver injury and uveitis. However, research exploring their dual therapeutic effects within a single organism remains lacking, leaving the key active components and mechanisms of action largely uninvestigated.

This exploratory study aimed to establish a rat model of liver injury combined with uveitis and investigated its pathological mechanisms, evaluating the therapeutic efficacy of PVPs in alleviating liver injury combined with uveitis in rats. Additionally, it explored the mechanism of action and identified key active ingredients of PVPs, offering potential new directions for the development of clinical therapeutic drugs.

A rat model of liver injury with uveitis was established through intraperitoneal d-GalN/LPS injection. Metabolomics and proteomics were applied to investigate pathological mechanisms, followed by validation using acylcarnitine and S100A9 inhibitors. PVPs were administered to evaluate therapeutic effects and explore mechanisms involved in alleviating liver injury and uveitis. Network pharmacology combined with molecular docking identified critical active components in PVPs. Subsequent animal experiments verified the efficacy of the representative component in improving liver injury and uveitis.

d-GalN/LPS (150 mg/kg : 1 mg/kg) induced significant liver injury and uveitis in rats. Metabolomics analysis pointed to acylcarnitine as a key metabolite, and its inhibition reduced inflammation. Proteomics analysis implicated S100A9 in inflammation and immunity. Then, we intervened with S100A9 inhibitors in the model rats. The results suggested that the pathological mechanism of liver injury and uveitis caused by d-GalN/LPS involved the upregulation of S100A9 expression, an increase in PP2A activity, the inhibition of AMPK phosphorylation, and the downregulation of CPT1A, leading to the accumulation of acylcarnitine and promoting the inflammatory response in the liver and retina. Further, experiments involving PVPs demonstrated dose-dependent improvements in liver injury and uveitis caused by d-GalN/LPS. The underlying mechanism of action involved suppression of S100A9 expression, reduction of PP2A activity, activation of AMPK, upregulation of CPT1A, and subsequent reduction in acylcarnitine accumulation in both the liver and retina. This mechanism effectively alleviated the inflammatory effects induced by d-GalN/LPS. Network pharmacology and molecular docking analyses pinpointed several key active components of PVPs-namely, rosmarinic acid, salviaflaside, esculetin, 2-hydroxycinnamic acid, 3,4-dihydroxybenzaldehyde, and 7,8-dihydroxycoumarin-that play significant roles in mitigating liver injury and uveitis. Follow-up experiments using the representative active component rosmarinic acid in rats confirmed its efficacy in improving symptoms of d-GalN/LPS-induced liver injury and uveitis, further validating the therapeutic potential of these key active components.

This study successfully established a rat model of liver injury combined with uveitis and confirmed the efficacy of PVPs in alleviating this condition. Furthermore, it determined that the underlying mechanism involves regulation of the S100A9-PP2A-AMPK pathway, with rosmarinic acid identified as a key active compound. These findings provide a basis for clinical studies on liver-eye comorbidities and offer critical evidence for further research and drug development of PVPs in liver-clearing and eye-brightening.

Because liver fibrosis causes several insults that can result in death, it is regarded as an epidemic health issue. As "an inhibitor of the sodium-glucose cotransporter-2 (SGLT2)," Dapagliflozin (Dapa) is one of the newest anti-diabetic drugs used to treat type 2 diabetes mellitus. Dapa's antioxidant, anti-inflammatory, and antifibrotic properties produced positive impacts in numerous human and animal models. Due to Dapa's previously documented properties, we planned this investigation to elucidate the protective function of Dapa in male rat liver fibrosis caused by thioacetamide (TAA) as well as the expected pathways.

There were four groups of 24 rats: a control group, a TAA group that received (100 mg/kg b.wt intraperitoneally twice a week for 6 weeks), "TAA + Dapa" groups that given oral Dapa at (1 and 2 mg/kg b.wt. for 4 weeks in addition to TAA injections).

It was shown that TAA injections increased toll-like receptor 4 (TLR4) (509.6%), tumor necrosis factor (TNF-α) (298.8%), alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), interleukin-6 (IL-6) (330.9%), phosphotidylinositol-3-kinase (PI3K) (428.9% %), and transforming growth factor-beta (TGF-β1) (416.6%) levels. All of these markers were considerably reduced by Dapa treatment. In addition, reduced glutathione (GSH), nuclear factor erythroid 2-related factor 2 (Nrf2) (79%), albumin, Heme-oxygenase 1 (HO-1) (69%), and superoxide dismutase (SOD) were all decreased after TAA injection; however, they were restored by Dapa administration. The Dapa-treated groups had higher Nrf2 and HO-1 gene expressions, based on the results of PCR. Biochemical outcomes were validated by histopathological results. Immunohistopathological study revealed that DAPA treatment decreased caspase-3 and alpha-smooth Muscle Actin (αSMA) expression.

Due to its interactions with the Nrf2/HO-1 and TLR4 pathways, our research showed that Dapa had antioxidant and anti-inflammatory qualities against TAA-induced liver fibrosis.

To investigate the diagnostic value of spectral CT in calculating extracellular volume fraction (ECV) for assessing the severity of liver cirrhosis. This retrospective study enrolled 172 participants, including 127 patients diagnosed with liver cirrhosis and 45 matched controls, all of whom underwent spectral CT hepatic enhancement imaging. Disease severity stratification was performed using the Child-Pugh classification system. ECV values were derived from the iodine density map during the delayed phase. These ECV values were then compared across the control group and subclassified cirrhosis groups (Child-Pugh classes A-C). Furthermore, a correlation analysis was performed to assess the relationship between ECV values and Child-Pugh scores in liver cirrhosis. Receiver operating characteristic (ROC) curves were constructed to evaluate the diagnostic performance of ECV values and MELD-Na in the Child-Pugh classification of liver cirrhosis. The ECV values were 25.49 ± 3.15, 29.73 ± 3.20, 35.64 ± 3.15, and 45.30 ± 5.16 for the control, Child-Pugh A, Child-Pugh B, and Child-Pugh C group, respectively, demonstrating significant intergroup differences (F = 184.67 P < 0.001). A strong positive correlation was observed between ECV and Child-Pugh liver function classification (r = 0.791, P < 0.001). The diagnostic performance of ECV for differentiating between Child-Pugh classes A and B (AUC: 0.901), B and C (AUC: 0.966) was higher compared to the MELD-Na score (AUC: 0.772 and 0.868) (P < 0.05, respectively). Multivariate analyses showed that ECV was an independent factor for cirrhosis (OR 1.610, P < 0.001). ECV values measured using spectral CT can serve as a noninvasive biomarker for assessing the severity of liver cirrhosis.

Jatropha tanjorensis has been employed to treat various diseases. This study aimed to investigate the mode of action of J. tanjorensis against liver fibrogenesis induced by carbon tetrachloride (CCl4). Liver fibrosis was induced in male Wistar rats by exposing them to CCl4 for nine weeks. From weeks 7 to 9, J. tanjorensis extract was administered. The study evaluated the activities of antioxidant enzymes and lipid peroxidation (LPO) levels, expression of TGFβ1 and fibrosis-associated proteins, and liver histology. J. tanjorensis significantly increased the depleted antioxidant activities induced by CCl4 while significantly reducing the elevated LPO levels caused by CCl4. Furthermore, it downregulated the upregulation of TGFβ1 and fibrosis-associated proteins induced by CCl4 and mitigated the liver lesions and collagen deposits resulting from CCl4 exposure. Overall, J. tanjorensis alleviated liver injury and fibrogenesis by exerting its antioxidant effect and inhibition of TGFβ1.

Liver diseases, such as hepatitis, cirrhosis, and liver cancer, pose significant public health challenges, ranking as the twelfth leading cause of death globally. Given the liver's critical functions in metabolism, detoxification, and biosynthesis, its impairment can lead to severe consequences, often resulting in end-stage liver failure. Although liver transplantation is regarded as the definitive intervention for advanced liver disease, factors such as a shortage of donors and potential surgical complications necessitate the investigation of non-surgical regenerative medicine alternatives. This manuscript provides a comprehensive review of innovative non-surgical therapies aimed at liver regeneration, with an emphasis on both cell-based and cell-free approaches. It examines the contributions of various stem cell populations, including mesenchymal stem cells, hematopoietic stem cells, and induced pluripotent stem cells, in facilitating liver repair through mechanisms of differentiation and paracrine signaling. Furthermore, it explores the therapeutic potential of exosomes and conditioned media derived from stem cells as biotherapeutic agents in the context of regenerative medicine. By elucidating the mechanisms that underpin liver regeneration, this study aspires to inform the development of effective therapeutic strategies to address liver diseases and slow their progression. By elucidating the underlying mechanisms of liver regeneration, this study aims to contribute to the development of effective therapeutic strategies to address liver diseases and slow their progression.

Liver fibrosis is an over-reacted wound healing that becomes lethal in its late stage, when hepatic stellate cells (HSCs) trigger fibrotic response, proliferation of connective tissue and build-up of directional fibrous tissue bands (septa). Current in vitro models of liver fibrosis cannot reproduce liver lobule structure and the dynamic formation of septa at the same time, and the known biochemical cues underlying the progression of liver fibrosis cannot explain directional formation of fibrotic tissue. Here we report a microfabricated in vitro model that reproduces both the hexagonal liver lobule structure and the dynamic directionality of septa formation. By using collagen and primary mouse HSCs or human HSC lines, we found that tension was necessary to coordinate the cell migration that contributes to the band-like cell distribution and that HSCs sensed directional biophysical cues through liquid-liquid phase separation. This system allows the study of the biophysical interaction of HSCs and collagen during the formation of septa structures, and could be used to deepen our understanding of liver fibrosis progression.

The ameliorative effect of traditional Chinese medicine (TCM) on hepatic fibrosis has been widely recognized and researched, but studies on the mechanism of action have been hampered by its complex composition, which requires more in-depth studies to elucidate why and how TCM works. The theory of TCM believes that the liver is closely related to blood circulation, and hepatic fibrosis is caused by blood stagnation. Taohong Siwu Decoction (THSW) is a classic formula for nourishing and invigorating blood and has been used clinically for centuries. Current evidence has demonstrated its ameliorative effect on hepatic fibrosis, but the exact mechanism of action remains unclear.

Exploring the possible mechanism of the anti-hepatic fibrosis effect of THSW by proteomics and validating with in vivo and in vitro studies.

The carbon tetrachloride (CCl4)-induced fibrosis model was conducted in mice and treated with THSW in vivo with colchicine as the positive control. Then serum biomarker alanine aminotransferase (ALT), aspartate aminotransferase (AST), and histopathological analysis were evaluated to examine the effects of THSW. And hepatic fibrosis indicators alpha-smooth muscle actin (α-SMA) and Collagen Ⅰ (Col-Ⅰ) were detected by western blotting, immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Additionally, the 4D Label-free quantitative proteomic analysis of liver samples was applied. In vitro, erastin-induced BRL-3A cells, a rat hepatocyte line, were performed as a hepatocyte ferroptosis model and treated with or without drug-containing serum of THSW. Finally, molecular docking was used to verify the binding ability of the main components of THSW to potential targets.

THSW treatment significantly ameliorated serum ALT, AST, hydroxyproline (Hyp) content, α-SMA and Col-Ⅰ mRNA expression in fibrosis mice. Further results showed that THSW decreased the malondialdehyde (MDA) and 4-Hydroxynonenal (4-HNE) content and increased the glutathione (GSH) content of liver tissue. Notably, proteomic analyses have identified 294 differentially expressed proteins in the THSW-treated group compared to the model group, with 97 proteins up-regulated and 197 down-regulated. Functional analysis of these differential proteins highlights the significant roles of inflammation and oxidative stress. Further validation in vivo and in vitro, THSW significantly improved the protein expression of glutathione S-transferase M1 (GSTM1), down-regulate the expression of transferrin receptor (TFRC), and kelch-like ECH-associated protein 1(Keap1) proteins, and promote the metabolism of GSH. Especially it reduced serum iron levels, increased total iron binding capacity, and up-regulated recombinant solute carrier family 7, member 11 (SLC7A11), nuclear factor erythroid 2-related factor 2 (Nrf2), and glutathione peroxidase 4 (GPX4) protein expression, suggesting the inhibition of hepatocyte ferroptosis. In addition, the molecular docking results showed that its main components, amygdalin, hydroxysafflor yellow A, paeoniflorin, and albiflorin, possessed good binding ability with Keap1.

THSW represents a novel therapeutic effect on hepatic fibrosis in mice, accompanied by inhibiting hepatocyte ferroptosis. Mechanically, THSW may regulate the glutathione metabolic pathway and TFRC expression through its main ingredients, such as amygdalin, hydroxysafflor yellow A, paeoniflorin, and albiflorin, thereby inhibiting hepatocyte ferroptosis.