Project description:Hepatic stellate cells(HSCs) are the main effector cells of liver fibrosis. In order to study the effect of mesenchymal stem cells(MSCs) on microRNAs expression of HSCs, we co-cultured HSCs LX-2 activated by TGFβ1 with human umbilical cord MSCs(hUC-MSCs) for 48 hours, and compared the differentially expressed miRNA with LX-2 cultured alone by high-throughput sequencing. The results showed that two mature microRNAs expressed increased, and nine expressed decreased.
Project description:Expression data from the hepatic stellate cell line LX-2 after treatment with the prolylhydroxylase inhibitor dimethyloxalylglycine (DMOG) and from the liver sinusoidal endothelial cell line TRP3 after incubation with conditioned medium of DMOG-treated LX-2 Prolyl-hydroxylase inhibitors such as dimethyloxalylglycine (DMOG) stabilize HIF-1α, thereby chemically inducing hypoxia, which also accelerates liver volume increase when given with portal rerouting. We used microarrays to clarify the cellular crosstalk of the different cell types in accelerated liver regeneration by examining the role of hepatic stellate cells (HSC) and liver sinusoidal endothelial cells (LSEC).
Project description:Hepatic fibrosis is the strongest contributor to hepatocarcinogenesis in metabolic dysfunction-associated steatotic liver disease (MASLD); however, the underlying mechanisms have yet to be fully elucidated. In 94 human MASLD biopsy samples, artificial intelligence-based morphological phenotyping of hepatic fiber and multi-omics analyses revealed that insulin growth factor-binding protein 7 (IGFBP-7) secreted from senescent periportal endothelial cells might transform stellate cells into a hepatocarcinogenesis-promoting phenotype. To test the effect of IGFBP-7 on HSC, a hepatic stellate cell line, LX-2, was cultured with recombinant IGFBP-7 (100ng/mL), resulting in their transformation to a more activated form than the control.
Project description:The purpose of this study was to identify the changes in gene expression that occur in LX-2 human hepatic stellate cells in response to depletion of mannose phosphate isomerase enzymatic activity.
Project description:To investigate the role of AEBP1 involved in hepatic stellate cells (HSCs), we inhibited AEBP1 expression by specific siRNA in human HSC line LX-2 cells. Total RNA was loaded for bulk RNA sequencing.
Project description:We found that the number of tumor-infiltrating myofibroblasts was positively correlated to tumor acidification status in hepatocellular carcinoma (HCC). Hepatic stellate cells (HSCs), the predominant precursors of liver myofibroblasts, were activated and transdifferentiated into myofibroblasts under acidic culture condition. To identify the molecular phenotype of LX-2 cells in acidic culture conditions, we further conducted a gene expression profile analysis. LX-2 cells cultured in pH 7.2 or pH 6.2 medium separately for six days was used in gene expression microarray analysis.
Project description:Liver fibrosis, characterized by excessive extracellular matrix deposition, is driven by activated hepatic stellate cells (HSCs). Due to the limited availability of anti-fibrotic drugs, research con-tinues to explore potential therapeutic agents. Moringa oleifera Lam. (MO), known for its various bioactive properties, is being investigated for its anti-fibrotic potential. This study focused on 1-phenyl-2-pentanol (1-PHE), a compound derived from MO leaves, and its impact on LX-2 hu-man hepatic stellate cell activation. TGF-β1-stimulated LX-2 cells were treated with MO extract or 1-PHE, and liver fibrosis markers were assessed at both gene and protein levels. Proteomic analysis and molecular docking were employed to identify potential protein targets and signaling pathways affected by 1-PHE. 1-PHE treatment downregulated fibrosis markers, including colla-gen type I alpha 1 chain (COL1A1), collagen type IV alpha 1 chain (COL4A1), mothers against decapentaplegic homolog 2 and 3 (SMAD2/3), and matrix metalloproteinase-2 (MMP2), and re-duced the secretion of matrix metalloproteinase-9 (MMP9). Proteomic analysis revealed the pos-sible mechanism of 1-PHE in modulating the Wnt/β-catenin pathway. These findings suggest that 1-PHE may suppress HSCs activation by inhibiting the TGF-β1 and Wnt/β-catenin signaling pathways, indicating its potential as an anti-liver fibrosis agent. Further research is warranted to validate these findings.
Project description:Purpose: We recently identified 39 human microRNAs, which effectively suppress hepatitis B virus (HBV) replication in hepatocytes. Because chronic HBV infection often results in active, hepatitis-related liver fibrosis, we assessed whether any of these microRNAs have anti-fibrotic potential and predicted that miR-6133-5p may target several fibrosis-related genes. Methods: The hepatic stellate cell line LX-2 was transfected with miR-6133-5p mimic and subsequently treated with TGF-β. mRNA and protein products of COL1A1, encoding collagen, and ACTA2, an activation marker of hepatic stellate cells, were quantified. Results: Expression of COL1A1 and ACTA2 was markedly reduced in LX-2 cells treated with miR-6133-5p. Interestingly, phosphorylation of JNK also was significantly decreased by miR-6133-5p treatment. The expression of several predicted target genes of miR-6133-5p, including TGFBR2 and FGFR1, was also reduced in miR-6133-5p-treated cells. The knockdown of TGFBR2 by the corresponding small interfering RNA greatly suppressed the expression of COL1A1 and ACTA2. Treatment with the JNK inhibitor, SP600125, also suppressed COL1A1 and ACTA2 expression, indicating that TGFBR2 and JNK would mediate the anti-fibrotic effect of miR-6133-5p. Downregulation of FGFR1 may result in a decrease of phosphorylated AKT, ERK, and JNK. Conclusions: miR-6133-5p has a strong anti-fibrotic effect, mediated by inactivation of TGFBR2, AKT and JNK.