长链非编码RNA H19在小鼠肝纤维化模型中的表达变化

In this paper, Background and Purpose: Liver fibrosis is the pathological process of abnormal hyperplasia of connective tissue in the liver and the major pathological manifestations are excessive deposition of extracellular matrix(ECM). Liver fibrosis is the common pathological process of many liver diseases. At the same time, it is also the only reversible process, indicating that the treatment for liver fibrosis is the key to the prevention of liver diseases. Long noncoding RNA (lncRNA) is a transcription of the length more than 200nt RNA. H19 is one of the earlier found lncRNA associated with disease. Many reports demonstrate that H19 in HCC is increased. Overexpression of H19 resultes in increaseing liver cancer cell proliferation, whereas knockdown of H19 leads to proliferation and invasion of hepatocellular carcinoma cells decreased significantly. However, the expression of H19 in liver fibrosis remains elusive. This paper aims to determine the expression of lncRNA H19 in the mice models of liver fibrosis, and to provide an experimental basis for functional studies. Methods In vivo experiments: C57BL/6 mice were taken to give CCl4 by intraperitoneal injection to create liver fibrosis models.These models were identified by detecting the serum ALT / AST levels, pathological HE and Masson staining. QRT-PCR method was taken to detect the expression level of H19 and other genes related to liver fibrosis in mouse liver. In vitro experiments: We chose 400 mM ethanol and 400 μM H2O2 to stimulate HL7702 cells to build hepatocyte injury models. In order to establish the in vitro model of hepatic fibrosis, the hepatic stellate cell line LX-2 cells were cultured in the presence of 5 ng/ml TGF-β1 for 48h to be activated.QRT-PCR method was taken to explore the expression level of H19 in HL7702 cells and LX-2 cells. Results In vivo experiments: Pathological HE and Masson staining results indicated that the mice of liver fibrosis model had their liver tissue collagen fibers hyperplasia. Compared with the control group, the mice of liver fibrosis model significantly elevated serum ALT levels, demonstrating that the liver of liver fibrosis model group had been damaged and the models had been succeed. QRT-PCR results showed that the Timp1、Col1α1 and H19 expression level of CCl4 model in liver were increased significantly. In vitro experiments: The expression level of H19 in HL7702 cells didn't change significantly when they were treated by ethanol or H2O2 solution for 2 h while the expression level of H19 was significantly increased in hepatic stellate cells (LX-2) after they were activated by TGF-β1. Conclusion The expression level of H19 was increased in mice liver fibrosis model and in LX-2 cells when they were activated.