Effect of cobra venom NGF on inducing apoptosis of LX2 cells by inhibiting Akt signaling pathway and its mechanism / 中国药理学通报

Aim To study the effect of cobra venom nerve growth factor ( NGF) on inducing the apoptosis of LX2 cells, the key cells of hepatic fibrosis , through Akt signaling pathway and its underlying mechanism .Methods CCK-8 method was used to detect the pro-liferation of LX2 cells at different concentrations of NGF and LY294002 .Flow cytometry was applied to detect the effect of NGF on the apoptosis of LX 2 cells. Western blot was used to study the effects of NGF and LY294002 respectively , and their combination on the p-Akt protein level .Results NGF could decrease the survival rate of LX2 cells, and the minimum effective concentration was 1mg· L-1; it increased the apopto-sis rate of LX2 cells within the rise of concentration un-der a certain of range and decreased the expression level of p-Akt, but it had no significant effect on the ex-pression of Akt .Conclusions NGF may promote the apoptosis of LX2 cells by inhibiting the activation of PI3K/Akt signaling pathway in a concentration-de-pendent manner .The study of the pathogenesis of liver fibrosis is significant for the clinical treatment of liver fibrosis.

Effects on Proliferation Inhibition, Apoptosis and Secretion of Extracellular Matrix Function of Human Hepatic Stellate Cell by Plumbagin / 世界科学技术-中医药现代化

This article was aimed to study the effects of plumbagin to human hepatic stellate cells.Observations were made on the influence of proliferation inhibition rate,apoptosis,secretion of extracellular matrix function and matrix metalloproteinase (MMP) by plumbagin.HSC-LX2 and drug were co-incubated for 48 hours.Then,MTT assay was used in the detection of inhibition of cell proliferation.The flow cytometry was used in the detection of apoptosis.Immunohistochemical method was used to observe typeⅠ and Ⅲ collagen,MMP-1 and MMP-13 expression location and area.The results showed that the low,medium and high concentrations of plumbagin inhibited cell proliferation rate of HSC-LX2,induced apoptosis of cells,reduced the secretion of typeⅠ and Ⅲ collagen,and increased the secretion ability of MMP-1 and MMP-13.Effects mentioned above were dose-dependent with statistical difference (P < 0.05).Effects in the medium and high concentrations groups were stronger than colchicine group.It was concluded that plumbagin had the ability to inhibit cell proliferation rate of HSC-LX2,induce apoptosis,reduce the secretion of extracellular matrix,and increase the secretion ability of fibrin degradation enzyme.Therefore,it had intervention effect on the process of liver fibrosis.All effects mentioned above were dose-dependent.And effects in the medium and high concentrations groups were stronger than colchicine group.

Effects of hammerhad ribozyme targeting connective tissue growth factor on collagen Ⅰ synthesis and cell cycle progression of human hepatic stellate cells / 吉林大学学报(医学版)

Objective To study the inhibitory effect of hammerhead ribozyme targeting connective tissue growth factor(CTGF) on collagen I synthesis and cell cycle progression of human hepatic stellate cell line(LX-2) cells.Methods Hammerhead ribozyme cDNA targeting CTGF mRNA plus two self-cleaving sequences were inserted into pTriEx2 vector to construct a recombinant vector pTriCTGF-Rz.LX-2 cells were transfected with either pTriEx2 or pTriCTGF-Rz and further stimulated with or without TGF-1.There were five groups in the experiment:control group,pTriEx2 group,pTriCTGF-Rz group,pTriEx2 plus TGF-?1 group,and pTrCTGF-Rz plus TGF?1 group.Semi-quantitative RT-PCR was used to detect the levels of CTGF mRNA and collagen Ⅰ mRNA.ELISA and flow cytometry were used to detect the levels of collagen Ⅰ secretion and cell cycle.Results Transfection of pTriCTGF-Rz into LX-2 cells reduced the CTGF mRNA and collagen Ⅰ mRNA levels as well as collagen Ⅰ protein level compared with pTriEx2 group(P