Study on the Effects and Mechanism of Matrine on Proliferation and Collagen Synthesis of Hepatic Stellate Cells CFSC-8B Activated by Acetaldehyde / 中国药房

ABSTRACT

OBJECTIVE：To study the effects of matrine on proliferation and collagen synthesis of rat hepatic stellate cells CFSC-8B activated by acetaldehyde ，and to investigate its possible mechanism. METHODS ：CFSC-8B cells cultured in vitro were divided into blank control group ，model group ，positive control group （2.5 μmol/L colchicine）and matrine low ，medium and high concentration groups （30，60，120 μmol/L）. Except for blank control group ，other groups were activated with 200 μmol/L acetaldehyde for 24 h；medicine groups were intervened with relevant medicine for 24 h（blank control group and model group were intervened with equal volume blank medium ）. Survival rate of cell was detected by CCK- 8 assay. C ells were divided into blank control group ，model group ，positive control group （2.5 μmol/L colchicine），matrine medium and high concentration groups （60，120 μmol/L），then activated and treated with same method. Hydroxyprolin （Hyp）content in cell culture solution was tested by enzyme digestion. The contents of Col- Ⅰ and Col- Ⅲ in cell culture solution were determined by ELISA. mRNA expressionss of α-SMA，TGF-β1，TβR-І，TβR-Ⅱ，Smad3，Smad4 and Smad 7 in cells were detected by RT-PCR. The protein expressions of α-SMA，TGF-β1，TβR-Ⅰ，TβR- Ⅱ，Smad3，Smad4 and Samd 7 in cells were detected by Western blotting. RESULTS ：Compared with blank control group ，survival rate of cells in model group was increased significantly （P＜0.05）；the contents of Hyp ，Col-Ⅰ and Col- Ⅲ in cell culture solution ，mRNA and its protein expressions of α-SMA，TGF-β1，TβR-Ⅰ，TβR-Ⅱ，Smad3，Smad4 in cells were increased significantly in model group （P＜0.05），while the mRNA and protein expression of Smad 7 was decreased significantly（P＜0.05）. Compared with model group ，survival rate of cells ，the contents of Hyp ，Col-Ⅰ and Col- Ⅲ in cell culture solution，the mRNA and protein expressions of α-SMA and Smad 4 were decreased significantly in positive control group and matrine medium and high concentration groups （P＜0.05）， while the mRNA and protein expression of Smad 7 was WF-0099） increased significantly （P＜0.05）；the mRNA and proteinexpressions of TGF-β1，TβR-Ⅰ，TβR-Ⅱ and Smad 3 were decreased significantly in positive control group and matrine high concentration group （P＜0.05）. Compared with matrine medium concentration group ，all above indexes were improved significantly in matrine high concentration group （P＜0.05）. CONCLUSIONS ：Matrine can suppress the proliferation and collagen synthesis of CFSC- 8B cells activated by acetaldehyde ，with a centain concentrlation dependence ，the mechanism of which may be associated with regulating the conduction of TGFβ/Smad signal pathway.