• Humans
• Female
• Carcinoma, Intraductal, Noninfiltrating/pathology
• Spheroids, Cellular/metabolism
• Breast Neoplasms/pathology
• Calcinosis/metabolism
• Minerals
• Apatites
• Tumor Microenvironment

• Calcium oxalate crystallization
• Forkhead box protein O1
• Kidney stone
• Vitamin D receptor
• microRNA-484

• antioxidant
• antiurolithiasis
• sodium oxalate
• sulfated galactans
• sulfation

MicroRNAs (miRNAs or miRs) are critical regulators in various diseases. In the current study, the role of miR-30c-5p in the formation of sodium oxalate-induced kidney stones was investigated. For this purpose, human renal tubular epithelial cells (HK-2 cells) were incubated with sodium oxalate at the concentrations of 100, 250, 500, 750 and 1,000 µM. Cell viability and the miR-30c-5p expression level were respectively measured by CCK-8 assay and RT-qPCR. After separately transfecting miR-30c-5p mimic and inhibitor into the HK-2 cells, the cell apoptotic rate, the levels of mitochondrial membrane potential (MMP) and ROS were determined by flow cytometry. The levels of oxidative stress indicators [lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT)] were determined using commercial kits. Crystal-cell adhesion assay was performed to evaluate the crystal adhesion capacity in vitro. miR-30c-5p binding at autophagy related 5 (ATG5) was predicted by TargetScan7.2 and further verified by dual-luciferase reporter assay. Rescue experiments were performed to confirm the molecular mechanisms underlying sodium oxalate-induced kidney formation in HK-2 cells. The results revealed that sodium oxalate decreased the viability of HK-2 cells in a concentration-dependent manner, and that miR-30c-5p expression was significantly downregulated by exposure to 750 µM sodium oxalate. In addition, the increase in cell apoptosis and crystal number, and the upregulated levels of LDH, MDA and ROS were reversed by the overexpression of miR-30c-5p. Moreover, the overexpression of miR-30c-5p upregulated the levels of SOD, CAT and MMP induced by sodium oxalate. ATG5 was directly regulated by miR-30c-5p, and the inhibition of cell cytotoxicity and crystal-cell adhesion induced by miR-30c-5p mimic was blocked by ATG5. These data indicated that the overexpression of miR-30c-5p alleviated cell cytotoxicity and crystal-cell adhesion induced by sodium oxalate through ATG5. Thus, the current study provides a better understanding of the role of miR-30c-5p in sodium oxalate-induced kidney stones.

• TGF-β1
• epithelial to mesenchymal transition
• osteogenic differentiation
• oxalate
• renal medulla

• Apoptosis
• Berberine
• High glucose
• NF-E2-related factor 2
• Phosphatidylinositol 3-kinase (PI3K)/Akt
• Renal tubular epithelial cells

The present study aimed to investigate the mechanism of hepatoprotective effect of Erzhi Pill (EZP) on the liver injury via observing TSC/mTOR signaling pathway activation. The experimental liver injury was induced by 2-acetylaminofluorene (2-AAF) treatment combined with partial hepatectomy (PH). EZP treated 2-AAF/PH-induced liver injury by the therapeutic and prophylactic administration. After the administration of EZP, the activities of aspartic transaminase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AKP), and gamma-glutamyl transpeptidase (γ-GT) were decreased, followed by the decreased levels of hepatocyte apoptosis and caspase-3 expression. However, the secretion of albumin, liver weight, and index of liver weight were elevated. Microscopic examination showed that EZP restored pathological liver injury. Meanwhile, Rheb and mammalian target of rapamycin (mTOR) activation were suppressed, and tuberous sclerosis complex (TSC) expression was elevated in liver tissues induced by 2-AAF/PHx and accompanied with lower-expression of Bax, Notch1, p70S6K, and 4E-EIF and upregulated levels of Bcl-2 and Cyclin D. Hepatoprotective effect of EZP was possibly realized via inhibiting TSC/mTOR signaling pathway to suppress excessive apoptosis of hepatocyte.

• Calcium oxalate
• Madin–Darby canine kidney cells
• Phytotherapy
• Renal epithelial cells
• Terminalia arjuna
• Urolithiasis

Kidney stone formation during hyperoxaluric condition is inherently dependent on the interaction between renal epithelial cells and calcium oxalate (CaOx) crystals. Although modern medicine has progressed in terms of removal of these stones, recurrence and persistent side effects restricts their use. Strategies involving plant based agents which could be used as adjunct therapy is an area which needs to be explored. Plant proteins having antilithiatic activity is a hitherto unexplored area and therefore, we conducted a detailed identification and characterization of antilithiatic proteins from Terminalia arjuna (T. arjuna). Proteins were isolated from the dried bark of T. arjuna and those having molecular weights > 3 kDa were subjected to anion exchange chromatography followed by gel filtration chromatography. Four proteins were identified exhibiting inhibitory activity against CaOx crystallization and crystal growth kinetics The cytoprotective and anti-apoptotic efficacy of these purified proteins was further investigated on oxalate injured renal epithelial cells (MDCK and NRK-52E) wherein, injury due to oxalate was significantly attenuated and led to a dose dependent increase in viability of these cells. These proteins also prevented the interaction of the CaOx crystals to the cell surface and reduced the number of apoptotic cells. Identification of these 4 anionic proteins from the bark of T. arjuna was carried out by Matrix-assisted laser desorption/ionization-time of flight Mass spectrometry (MALDI-TOF MS). This was followed by database search with the MASCOT server and sequence similarity was found with Nuclear pore anchor, DEAD Box ATP-dependent RNA helicase 45, Lon protease homolog 1 and Heat shock protein 90–3. These novel proteins isolated from T. arjuna have the potential to inhibit CaOx crystallization and promote cell survival and therefore, offer novel avenues which need to be explored further for the medical management of urolithiasis.

• Animals
• Anions
• Apoptosis/drug effects
• Calcium Oxalate/chemistry
• Calcium Oxalate/metabolism
• Cell Line
• Chromatography, Gel
• Chromatography, Ion Exchange
• Crystallization
• Dogs
• Epithelial Cells/cytology
• Epithelial Cells/drug effects
• Epithelial Cells/metabolism
• Kidney Tubules, Collecting/cytology
• Madin Darby Canine Kidney Cells
• Phytotherapy/methods
• Plant Bark/metabolism
• Plant Proteins/isolation & purification
• Plant Proteins/metabolism
• Plant Proteins/pharmacology
• Proteomics/methods
• Rats
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Terminalia/metabolism
• Urolithiasis/drug therapy

Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Hyperglycemia and high glucose in vitro also lead to apoptosis, a form of programmed cell death. High glucose similar to those seen with hyperglycemia in people with diabetes mellitus, lead to accelerated apoptosis, a form of programmed cell death characterized by cell shrinkage, chromatin condensation and DNA fragmentation, in variety of cell types, including renal proximal tubular epithelial cells.

• Tubular cells
• Renal
• Apoptosis
• Diabetes

Nephrolithiasis is a multi-factorial disease which, in the majority of cases, involves the renal deposition of calcium oxalate. Oxalate is a metabolic end product excreted primarily by the kidney. Previous studies have shown that elevated levels of oxalate are detrimental to the renal epithelial cells; however, oxalate renal epithelial cell interactions are not completely understood. In this study, we utilized an unbiased approach of gene expression profiling using Affymetrix HG_U133_plus2 gene chips to understand the global gene expression changes in human renal epithelial cells [HK-2] after exposure to oxalate. We analyzed the expression of 47,000 transcripts and variants, including 38,500 well characterized human genes, in the HK2 cells after 4 hours and 24 hours of oxalate exposure. Gene expression was compared among replicates as per the Affymetrix statistical program. Gene expression among various groups was compared using various analytical tools, and differentially expressed genes were classified according to the Gene Ontology Functional Category. The results from this study show that oxalate exposure induces significant expression changes in many genes. We show for the first time that oxalate exposure induces as well as shuts off genes differentially. We found 750 up-regulated and 2276 down-regulated genes which have not been reported before. Our results also show that renal cells exposed to oxalate results in the regulation of genes that are associated with specific molecular function, biological processes, and other cellular components. In addition we have identified a set of 20 genes that is differentially regulated by oxalate irrespective of duration of exposure and may be useful in monitoring oxalate nephrotoxicity. Taken together our studies profile global gene expression changes and provide a unique insight into oxalate renal cell interactions and oxalate nephrotoxicity.

Oxalate is a metabolic end product excreted by the kidney. Mild increases in urinary oxalate are most commonly associated with Nephrolithiasis. Chronically high levels of urinary oxalate, as seen in patients with primary hyperoxaluria, are driving factor for recurrent renal stones, and ultimately lead to renal failure, calcification of soft tissue and premature death. In previous studies others and we have demonstrated that high levels of oxalate promote injury of renal epithelial cells. However, methods to monitor oxalate induced renal injury are limited. In the present study we evaluated changes in expression of Kidney Injury Molecule-1 (KIM-1) in response to oxalate in human renal cells (HK2 cells) in culture and in renal tissue and urine samples in hyperoxaluric animals which mimic in vitro and in vivo models of hyper-oxaluria. Results presented, herein demonstrate that oxalate exposure resulted in increased expression of KIM-1 m RNA as well as protein in HK2 cells. These effects were rapid and concentration dependent. Using in vivo models of hyperoxaluria we observed elevated expression of KIM-1 in renal tissues of hyperoxaluric rats as compared to normal controls. The increase in KIM-1 was both at protein and mRNA level, suggesting transcriptional activation of KIM-1 in response to oxalate exposure. Interestingly, in addition to increased KIM-1 expression, we observed increased levels of the ectodomain of KIM-1 in urine collected from hyperoxaluric rats. To the best of our knowledge our studies are the first direct demonstration of regulation of KIM-1 in response to oxalate exposure in renal epithelial cells in vitro and in vivo. Our results suggest that detection of KIM-1 over-expression and measurement of the ectodomain of KIM-1 in urine may hold promise as a marker to monitor oxalate nephrotoxicity in hyperoxaluria.

• Animals
• Cell Adhesion Molecules/chemistry
• Cell Adhesion Molecules/genetics
• Cell Adhesion Molecules/metabolism
• Cell Adhesion Molecules/urine
• Cell Line
• Crystallization
• Epithelial Cells/drug effects
• Epithelial Cells/metabolism
• Epithelial Cells/pathology
• Hepatitis A Virus Cellular Receptor 1
• Humans
• Hyperoxaluria/genetics
• Hyperoxaluria/pathology
• Hyperoxaluria/urine
• Immunohistochemistry
• Kidney/drug effects
• Kidney/metabolism
• Kidney/pathology
• Kidney Tubules/drug effects
• Kidney Tubules/metabolism
• Kidney Tubules/pathology
• Male
• Membrane Glycoproteins/genetics
• Membrane Glycoproteins/metabolism
• Oxalates/pharmacology
• Protein Structure, Tertiary
• RNA, Messenger/genetics
• RNA, Messenger/metabolism
• Rats
• Rats, Wistar
• Receptors, Virus/genetics
• Receptors, Virus/metabolism
• Up-Regulation/drug effects

• R01 DK054084 NIDDK NIH HHS
• NIH-RO1-DK-54084 NIDDK NIH HHS