鳖甲提取物对抑制TGF-β诱导的大鼠肝星状细胞活化的影响  

Effect of Carapax Trionycis in Inhibiting Activation of Rat Hepatic Stellate Cell Induced by TGF-β

作　　者：熊莎[1] 高建蓉[2] 胡祖良[2] 刘焱文[1] 尤朋涛[1] 

机构地区：[1]湖北中医药大学中药资源与化学重点实验室,武汉430065 [2]浙江衢化医院,浙江衢化324004

出　　处：《中国实验方剂学杂志》2017年第19期155-159,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：浙江省自然科学基金项目(LY14H280001);浙江省公共项目(2016C37037)

摘　　要：目的:研究鳖甲提取物相对分子质量<6 k Da肽段(P6)对转化生长因子-β(transforming growth factor-β,TGF-β)诱导的大鼠肝星状细胞HSC-T6的活化增殖与肝纤维化(hepatic fibrosis,HF)相关基因表达的影响,初步探究其抗肝纤维化的作用机制。方法:应用透析法得到P6;细胞增殖与活性检测(CCK8)法检测P6对HSC-T6细胞增殖的影响;实时荧光定量聚合酶反应(Real-time PCR)检测HSC-T6细胞内α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA),I型胶原(collagen type I,Col I),基质金属蛋白酶抑制因子-1(tissue inhibitor of metalloproteinase-1,TIMP-1)以及基质金属蛋白酶(matrix metalloproteinase-2,MMP-2)的mRNA表达水平;蛋白免疫印迹法(Western blot)分析HSC-T6细胞内α-SMA与Col I蛋白的表达。结果:与空白组比较,P6对HSC-T6细胞存活率无明显影响,但却能显著抑制TGF-β诱导的HSC-T6细胞的增殖(P<0.05)。与TGF-β组比较,P6各浓度均能显著降低HSC-T6细胞中α-SMA,Col I,TIMP-1 mRNA的表达,增加MMP-2 mRNA的表达(P<0.05,P<0.01),明显降低α-SMA与Col I蛋白的表达。结论:P6能抑制TGF-β诱导的HSC-T6细胞的活化增殖,减少细胞外基质(extracellular matrix,ECM)生成,促进其降解,发挥抗肝纤维化作用。Objective： To investigate the effect of P6（〈6 kDa） on activation, proliferation and hepatofibrosis-related genes in rat hepatic stellate cell line HSC-T6 induced by transforming growth factor-β （TGF-β）, in order to explore the mechanism of anti-hepatofibrosis effect. Method： Dialysis was applied to get P6. Cell counting Kit 8 （CCK8） was performed to evaluate cell viability and P6 effect on HSC-T6 proliferation. The mRNA expressions of α-smooth muscle actin （α-SMA）, collagen type I （Col I）, tissue inhibitor of metalloproteinase-1 （TIMP-1） and matrix metalloproteinase-2 （MMP-2） were determined by Real-time PCR. Protein expressions of α-SMA and Col I were assessed by Western blot. Result： Compared with control group, P6 did not significantly affect cell viability, but suppressed TGF-β-induced HSC-T6 proliferation （P〈0.05）. Compared with TGF-β group, mRNA expressions of α-SMA, Col I and TIMP-1 were significantly decreased by P6 in TGF-β-induced HSC-T6, while MMP-2 mRNA expression was markedly increased （P〈0.05, P〈0.01）, α-SMA and Col I protein expressions were also decreased obviously. Conclusion： These results demonstrated that P6 had an anti-hepatofibrosis effect by inhibiting HSC-T6 activation and proliferation induced by TGF-β, reducing production and accelerating degradation of ECM.

关 键 词：鳖甲 HSC-T6 肝纤维化 活化 机制 

分 类 号：R285.5[医药卫生—中药学]