Study Of Mcchanism Of TRPM7Involved In Proliferation Of HSC-T6Induced By PDGF

Hepatic fibrosis is regarded as a pathologically repair reaction followed variouskinds of chronic liver diseases. It is characterized by an excessive deposition ofextracellular matrix proteins （ECM） of which type I collagen predominates. Hepaticfibrosis is a common stage of most chronic liver diseases regardless of the etiology, andits progression may lead to hepatic cirrhosis or hepatocelluar carcinoma （HCC）.Although hepatic fibrosis thought to be a reversible pathological state, there is noestablished effective therapy for hepatic fibrosis yet. Many anti-fibrosis drugs have beendeveloped in recent years. However, no effective anti-fibrotic therapies are availableuntil now. Accordingly, it is significance that understanding the molecularpathophysiology of hepatic fibrosis will lead to novel therapeutic strategies andanti-fibrotic drugs.TRPM7（transient receptor potential cation channel, subfamily M, member7） is anovel dual function protein that possesses both ion channel and protein kinase domains，as a central regulator of cell growth and proliferation.The topic is through to establish the hepatic fibrosis model by PDGF-induced, toget a preliminary understanding the effect of TRPM7on hepatic fibrosis. Meanwhilethrough detect PI3K/AKT signal transduction Pathway in the key signal transductionmo1ecules PI3K, AKT expression level changes in HSC, to explore the molecularmechanism of TRPM7on the progression of hepatic fibrosis. 1. The relation between TRPM7and proliferation of HSCEstablish liver fibrosis model induced by PDGF, and then treat with LY294002,which is the specific blockers of PI3K/AKT signaling pathway,2-APB、 Gd³⁺respectively, which are blockers of ion channels. The results showed that PDGF couldsignificantly stimulate the proliferation of HSC, but after LY294002、2-APB、Gd³⁺treatment, the proliferation is inhibited obviously.2. Effects of Liposomal transfection of TRPM7-siRNA on proliferation ofHSC-T6Observe the effect of TRPM7-siRNA on proliferation of HSC-T6by establishingTRPM7silence model. The results showed that TRPM7-siRNA could significantlyinhibit the proliferation of HSC; the inhibition rate is proportional to the concentrationof TRPM7-siRNA. As TRPM7expression reduced, α-SMA、Collagen mRNAexpression are also reduced. It showed that TRPM7-siRNA can inhibit activation andproliferation of HSC. The experiment discuss the effect of TRPM7on cell cycle ofHSC-T6, The research showed that TRPM7-siRNA can affect the proliferation ofHSC-T6，by promoting the accumulation of G₀/G₁period, reducing the number of G2/Mand S period.The results showed that TRPM7is related closely in the process of hepaticfibrosis.3. Effects of TRPM7on PI3K/AKT signaling pathway in HSC-T6cellEstablish liver fibrosis model induced by PDGF, and then treat with LY294002,which is the specific blockers of PI3K/AKT signaling pathway,2-APB、 Gd³⁺ respectively, which are blockers of ion channels. Compared with the normal group,PDGF can significantly stimulate HSC-T6TRPM7、α-SMA、Collagen mRNA andTRPM7、 p-PI3K、 p-AKT protein expression. After LY294002、2-APB、 Gd³⁺treatment,TRPM7、α-SMA、Collagen mRNA and TRPM7、p-PI3K、 p-AKT proteinexpression are inhibited obviously, but T-PI3K、T-AKT protein expression have nosignificant changes,To sum up, TRPM7-siRNA has anti-liver fibrosis effect, can be effective inimproving liver function, reducing the degree of hepatic fibrosis, Which might beassociated with the ability to regulate PI3K/AKT signaling pathway.