The Effect Of Mast Cell Chymase On Proliferation And Transdifferation Of Hepatic Stellate Cell

Objective:We used the technology of MTT、Western Blotting、RT-PCR to observe the expression of protein α-SMA、TGF-β1 and TGF-β1 mRNA、Col-I mRNA on the HSC by chymase. Investigate the effects of chymase on proliferation and transdifferentiation of HSC, and the effects on the development of the hepatic fibrosis.Methods:1.The Wistar male rats were anesthetized by the ether, then fixed the rat on the operating table, then opened the abdominal with "cross" cut to expose the liver, portal vein and vena cave.2.Then treated the liver with perfusion fluid, and we used the 3-4 cells what we had cultured to divide it into blank control group, lng/ml, 1Ong/ml and 100ng/ml chymase groups.3.we should to assay HSC proliferation by MTT and count the OD values.4. We sealed it with Western solution, then to make the target protein collected the antibody.5.We used the RT-PCR technology to detect the expression of TGF-β1 mRNA and Col-I mRNA.Results:1.Mast cell chymase promoted proliferation of HSC and within the observed range (lng/ml、10ng/ml、100ng/ml) it was showed concentration dependent, the higher the concentration of chymase, the higher of OD values, which was about the control group (no chymase group) 1.45 times (P<0.05).2.The results of Westren Blotting test showed:the expression of a-SMA and TGF-β1 were significantly increased on the HSC stimulated by chymase (P<0.05).3.The expression of the TGF-β1 mRNA and Col-I mRNA on HSC was significantly enhanced by RT-PCR technology, within the observed range it was showed concentration dependent (P<0.05).Conclusion:1.Chymase can significantly promote the proliferation of HSC, and it was depended the concentration of chymase.2. The expression of a-SMA and TGF-β1 was significantly increased in the HSC activation by chymase and in a concentration dependent manner.3.The expression of TGF-β1 mRNA and Col-I mRNA was significantly increased in the HSC activation by chymase and in a concentration dependent manner.4.The mast cell chymase can obvious activate HSC and promote it proliferation and transdifferentiation to myofibroblast.