Project description:Endoglin is a transmembrane receptor able to bind several members of the transforming growth factor-beta superfamliy. We used a siRNA-mediated knock down approach to analyse the function of Endoglin in hepatic stellate cells (HSC). We found that the targeted disruption is associated with significant changes in gene expression resulting in cellular changes that prolongated cellular activation, morphological maturation, transdifferentiation, expression of basement and fibrillar extracellular matrix proteins. In addition, the lack of Endoglin resulted in rearrangements of cytoskeletal organization. Keywords: gene function analysis, siRNA, gene knock down Overall design: We have performed a set of four microarray hybridizations using Agilent Whole Rat Genome Oligo Microarrays. We compartively analysed the gene expression profile in RNAs taken from: (I) untreated HSC taken from one animal vs. HSC that were taken from one animal and transfected with a control siRNA, (II) HSC taken from one animal and transfected with a siRNA targeting endoglin vs. HSC that were taken from one animal and transfected with a control siRNA, (III) untreated HSC that were pooled from three individual animals vs. HSC that were taken from three individual animals and independently transfected with siRNA control, and finally (IV) HSC that were taken from three animals and independently transfected with a siRNA targeting endoglin vs HSC that were taken from three individual animals and independently transfected with a siRNA control.
Project description:HSC-2 (hepatic stellate cells line from rat) were stably transfected with rno-miR-146a. Three different clones were selected (S1, S4, S5). We used Affymetrix rat genome RAT230 2.0 chip to monitor global transcriptome changes. Overall design: Two passages of each sample were used. Control - HSC-2 and GFPscr.
Project description:HSC-2 (hepatic stellate cells line from rat) were stably transfected with rno-miR-146a. Three different clones were selected (S1, S4, S5). We used Affymetrix rat genome RAT230 2.0 chip to monitor global transcriptome changes. Two passages of each sample were used. Control - HSC-2 and GFPscr.
Project description:Analysis of LBNF1 rat testes from controls, containing both somatic and all germ cell types and from irradiated rats in which all cells germ cells except type A spermatgogonia are eliminated. Results provide insight into distinguishing germ and somatic cell genes and identification of somatic cell genes that are upregulated after irradiation. Overall design: We studied 10 Sample replicates of control rat testes and 5 Sample replicates of irradiated rat testes.
Project description:Normal and cirrhotic LSECs can influence hepatic stellate cells (HSCs) differently. This project aims to determine the set of proteins secreted by LSECs from healthy and cirrhotic livers, and identify the protein(s) that promote HSC activation.
Project description:With the aim to reveal the function of microRNAs in Hepatic stellate cells (HSCs) in response to portal hypertension, primary rat HSCs were exposed to pressure (10mmHg, 1 h) by using a pressure induced apparatus. Appling next-generation sequencing screened the pressurization induced miRNAs expression profile in HSCs. Among them miR-9a-5p was confirmed to be significantly increased after loading pressure in HSC by RT-PCR. It was found that inhibition of miR-9a-5p could significantly restrain HSCs proliferation and activation under pressure overload. Moreover, the results showed that the induction of miR-9a-5p upon pressure load might by increasing the phosphorylation of Akt but not FAK and Erk1/2. Luciferase reporter assay and western blot suggested that Sirt1 was a potential target gene of miR-9a-5p. Finally, we revealed that miR-9a-5p level was apparently higher in rat liver fibrotic model than in the normal control while Sirt1 level was decreased in fibrotic liver tissue. In conclusion, our results suggest that miR-9a-5p regulate HSCs proliferation through negative regulation of Sirt1 and suggest a potential biomarker for portal hypertension. Overall design: Hepatic stellate cell microRNA profiles of 14-day activated pressure type and control were generated by deep sequencing
Project description:Activation and migration of hepatic stellate cells (HSCs) followed by matrix deposition are characteristics of liver fibrosis. Several studies have shown the importance of hepatocyte and endothelial cell-derived extracellular vesicles (EVs) in liver pathobiology. However, less is known about the role of HSC-derived EVs in liver diseases. In this study, we investigated the molecules released through HSC-derived EVs and whether these can promote fibrosis.
Project description:Adult-derived human liver stem/progenitor cells (ADHLSC) are obtained after primary culture of the liver parenchymal fraction. The cells are of fibroblastic morphology and exhibit a hepato-mesenchymal phenotype. Hepatic stellate cells (HSC) derived from the liver non-parenchymal fraction present a comparable morphology as ADHLSC. Because both ADHLSC and HSC are described as liver stem/progenitor cells, we strived to extensively compare both cell populations at different levels and to propose tools demonstrating their singularity. The database include full expression (HGU-219) measurements samples from 7 Adult-Derived Human Liver Stem/progenitor (n=7) and human hepatic stellate samples (n=7)