MicroRNA studies in in vitro and in vivo activated rat hepatic stellate cells: Emphasis on miR-146a
Ontology highlight
ABSTRACT:
ORGANISM(S): Caenorhabditis elegans Mus musculus Danio rerio Rattus norvegicus Homo sapiens Drosophila melanogaster
PROVIDER: GSE19463 | GEO | 2009/12/15
SECONDARY ACCESSION(S): PRJNA120333
REPOSITORIES: GEO
Similar Datasets
Project description:This SuperSeries is composed of the following subset Series: GSE19354: Expression data for rno-miR-146a overexpressing cell line HSC-2 GSE19462: microRNA profiling in vitro activated primary hepatic stellate cells Refer to individual Series
2010-05-18 | E-GEOD-19463 | biostudies-arrayexpress
Project description:MicroRNA studies in in vitro and in vivo activated rat hepatic stellate cells: Emphasis on miR-146a
| PRJNA120333 | ENA
Project description:AimTo understand which and how different miRNAs are implicated in the process of hepatic stellate cell (HSC) activation.MethodsWe used microarrays to examine the differential expression of miRNAs during in vitro activation of primary HSCs (pHSCs). The transcriptome changes upon stable transfection of rno-miR-146a into an HSC cell line were studied using cDNA microarrays. Selected differentially regulated miRNAs were investigated by quantitative real-time polymerase chain reaction during in vivo HSC activation. The effect of miRNA mimics and inhibitor on the in vitro activation of pHSCs was also evaluated.ResultsWe found that 16 miRNAs were upregulated and 26 were downregulated significantly in 10-d in vitro activated pHSCs in comparison to quiescent pHSCs. Overexpression of rno-miR-146a was characterized by marked upregulation of tissue inhibitor of metalloproteinase-3, which is implicated in the regulation of tumor necrosis factor-α activity. Differences in the regulation of selected miRNAs were observed comparing in vitro and in vivo HSC activation. Treatment with miR-26a and 29a mimics, and miR-214 inhibitor during in vitro activation of pHSCs induced significant downregulation of collagen type I transcription.ConclusionOur results emphasize the different regulation of miRNAs in in vitro and in vivo activated pHSCs. We also showed that miR-26a, 29a and 214 are involved in the regulation of collagen type I mRNA.
| S-EPMC3122263 | biostudies-literature
Project description:At present, no satisfactory anti-liver fibrosis drugs have been used clinically due to the poor targeting ability and short half-life period. This study aimed to explore the effects of a new TRAIL (TNF-related apoptosis-inducing ligand) preparation that can target aHSCs (activated hepatic stellate cells) on liver fibrosis and explain the possible underlying mechanism. Using our self-made drug carrier pPB-SSL that specifically targets aHSCs, recombinant human TRAIL (rhTRAIL) protein was embedded in (named as pPB-SSL-TRAIL) and applied to treat liver fibrotic mice as well as 3T3 fibroblast cells and aHSCs. Through in vitro and in vivo experiments, we found that, compared with the groups treated with TRAIL (free rhTRAIL) and SSL-TRAIL (rhTRAIL capsulated within unmodified liposome), the group treated with pPB-SSL-TRAIL nanoparticles showed significantly lower cell viability and higher cell apoptosis in vitro. The targeting delivering system pPB-SSL also significantly enhanced the anti-fibrotic effect, apoptosis induction and long circulation of rhTRAIL. After the treatment with pPB-SSL-TRAIL, apoptosis of aHSCs was notably increased and hepatic fibrosis in mice was remarkably alleviated. In vitro, pPB-SSL-TRAIL nanoparticles were mainly transported and located on membrane or into cytoplasm, but the particles were distributed mainly in mouse fibrotic liver and most on the cell membrane of aHSCs. In conclusion, rhTRAIL carried by pPB-SSL delivering system has prolonged circulation in blood, be able to target aHSCs specifically, and alleviate fibrosis both in vitro and in vivo. It presents promising prospect in the therapy of liver fibrosis, and it is worthwhile for us to develop it for clinical use.
| S-EPMC6378220 | biostudies-literature
Project description:BackgroundLiver fibrosis is characterized by the excessive formation and accumulation of matrix proteins as a result of wound healing in the liver. A main event during fibrogenesis is the activation of the liver resident quiescent hepatic stellate cell (qHSC). Recent studies suggest that reversion of the activated HSC (aHSC) phenotype into a quiescent-like phenotype could be a major cellular mechanism underlying fibrosis regression in the liver, thereby offering new therapeutic perspectives for the treatment of liver fibrosis. Whether human HSCs have the ability to undergo a similar reversion in phenotype is currently unknown. The aim of the present study is to identify experimental conditions that can revert the in vitro activated phenotype of primary human HSCs and consequently to map the molecular events associated with this reversion process by gene expression profiling.ResultsWe find that epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2) synergistically downregulate the expression of ACTA2 and LOX in primary human aHSCs. Their combination with oleic acid, palmitic acid, and retinol further potentiates a more quiescent-like phenotype as demonstrated by the abundant presence of retinyl ester-positive intra-cytoplasmic lipid droplets, low expression levels of activation markers, and a reduced basal as well as cytokine-stimulated proliferation and matrix metalloproteinase activity. Gene expression profiling experiments reveal that these in vitro reverted primary human HSCs (rHSCs) display an intermediary phenotype that is distinct from qHSCs and aHSCs. Interestingly, this intermediary phenotype is characterized by the increased expression of several previously identified signature genes of in vivo inactivated mouse HSCs such as CXCL1, CXCL2, and CTSS, suggesting also a potential role for these genes in promoting a quiescent-like phenotype in human HSCs.ConclusionsWe provide evidence for the ability of human primary aHSCs to revert in vitro to a transitional state through synergistic action of EGF, FGF2, dietary fatty acids and retinol, and provide a first phenotypic and genomic characterization of human in vitro rHSCs.
| S-EPMC4527231 | biostudies-literature
Project description:pHSCs were isolated from Wistar rats and cultivated in cell culture flasks. The miRNA was isolated at day 0 and 10 and hybridized onto the Ncode Multi-species miRNA microarray V2 from Invitrogen 3 samples from different rats were analyzed on miRNA microarray performing dye swap
2010-05-18 | E-GEOD-19462 | biostudies-arrayexpress
Project description:pHSCs were isolated from Wistar rats and cultivated in cell culture flasks. The miRNA was isolated at day 0 and 10 and hybridized onto the Ncode Multi-species miRNA microarray V2 from Invitrogen
2009-12-15 | GSE19462 | GEO
Project description:Hepatic stellate cells (HSC) are the major cellular drivers of liver fibrosis. Upon liver inflammation caused by a broad range of insults including non-alcoholic fatty liver, HSC transform from a quiescent into a proliferating, fibrotic phenotype. Although much is known about the pathophysiology of this process, exact cellular processes which occur in HSC and enable this transformation remain yet to be elucidated. In order to investigate this HSC transformation, we employed a simple, yet reliable model of HSC activation via an increase in growth medium serum concentration (serum activation). For that purpose, immortalized human LX-2 HSC were exposed to either 1% or 10% fetal bovine serum (FBS). Resulting quiescent (1% FBS) and activated (10% FBS) LX-2 cells were then subjected to in-depth mass spectrometry-based proteomics analysis as well as comprehensive phenotyping. Protein network analysis of activated LX-2 cells revealed an increase in the production of ribosomal proteins and proteins related to cell cycle control and migration, resulting in higher proliferation and faster migration phenotypes. Interestingly, we also observed a decrease in the expression of cholesterol and fatty acid biosynthesis proteins in accordance with a concomitant loss of cytosolic lipid droplets during activation. Overall, this work provides an update on HSC activation characteristics using contemporary proteomic and bioinformatic analyses and presents an accessible model for HSC activation. Data are available via ProteomeXchange with identifier PXD029121.
| S-EPMC8657869 | biostudies-literature
Project description:BACKGROUND: Hepatic stellate cells play a key role in the pathogenesis of hepatic fibrosis. AIMS: To examine the inhibitory effect of oestradiol on stellate cell activation. METHODS: In vivo, hepatic fibrosis was induced in rats by dimethylnitrosamine or pig serum. In vitro, rat stellate cells were activated by contact with plastic dishes resulting in their transformation into myofibroblast-like cells. RESULTS: In the dimethylnitrosamine and pig serum models, treatment with oestradiol at gestation related doses resulted in a dose dependent suppression of hepatic fibrosis with restored content of hepatic retinyl palmitate, reduced collagen content, lower areas of stellate cells which express alpha smooth muscle actin (alpha-SMA) and desmin, and lower procollagen type I and III mRNA levels in the liver. In cultured stellate cells, oestradiol inhibited type I collagen production, alpha-SMA expression, and cell proliferation. These findings suggest that oestradiol is a potent inhibitor of stellate cell transformation. CONCLUSION: The antifibrogenic role of oestradiol in the liver may contribute to the sex associated differences in the progression from hepatic fibrosis to cirrhosis
| S-EPMC1760074 | biostudies-other
Project description:Methionine adenosyltransferases (MATs) are critical enzymes that catalyze the formation of the methyl donor S-adenosyl methionine (SAM). The MAT2A gene, which encodes the catalytic subunit ?2, is induced in dedifferentiated liver. We previously demonstrated that MAT2A expression is enhanced in activated hepatic stellate cells (HSCs) and that silencing this gene reduces HSC activation. In this study, we examined the molecular mechanisms responsible for the transcriptional regulation of the MAT2A gene in HSCs. We identified peroxisome proliferator-activated receptor (PPAR) response elements (PPREs) in the rat MAT2A promoter. The PPAR? agonist rosiglitazone (RSG) promoted quiescence in the activated rat HSC cell line (BSC) or culture-activated primary rat HSCs, decreased MAT2A expression and promoter activity, and enhanced PPAR? binding to MAT2A PPREs. In vivo HSC activation in bile duct-ligated rats lowered PPAR? interaction with MAT2A PPREs. Silencing PPAR? increased MAT2A transcription, whereas overexpressing it had the opposite effect, demonstrating that PPAR? negatively controls this gene. Site-directed mutagenesis of PPREs abolished PPAR? recruitment to the MAT2A promoter and its inhibitory effect on MAT2A transcription in quiescent HSCs. PPRE mutations decreased the basal promoter activity of MAT2A in activated HSCs independent of PPAR?, indicating that other factors might be involved in PPRE interaction. We identified PPAR? binding to wild-type but not to mutated PPREs in activated cells. Furthermore, silencing PPAR? inhibited MAT2A expression and promoter activity. Forced expression of MAT2A in RSG-treated HSCs lowered PPAR? and enhanced PPAR? expression, thereby promoting an activated phenotype.We identified PPAR? as a negative regulator of MAT2A in quiescent HSCs. A switch from quiescence to activation abolishes this control and allows PPAR? to up-regulate MAT2A transcription.
| S-EPMC3342421 | biostudies-literature