Inhibitory Effects of Protopine on the Proliferation of Human Hepatic Stellate Cells HSC-LX 2 and Its Mechanism Study / 中国药房

ABSTRACT

OBJECTIVE： To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS： MTT assay was used to detect the effects of 25， 50， 100， 200， 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group （1640 medium containing 5％ fetal bovine serum）， protopine low-concentration， medium-concentration and high-concentration groups （100， 200， 400 μmol/L）. After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA， Collagen Ⅰ， Collagen Ⅲ， MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA， Collagen Ⅰ， Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS： The inhibitory rates of 25， 50， 100， 200， 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0， 6.9％， 18.7％， 34.2％， 48.9％， 53.9％， respectively. Compared with control group， mRNA expression of Collagen Ⅰ， TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration， medium-concentration and high-concentration groups， while protein expression of MMP- 2 was increased significantly， with statistical significance （P＜0.05 or P＜0.01）. Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups， mRNA expression of α-SMA and Collagen Ⅲ， protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS： Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation， and reduce the expression of a-SMA， Collagen Ⅰ， Collagen Ⅲ and TIMP-1， and increase the expression of MMP-2.