辣椒素调节肝星状细胞活性延缓肝纤维化的实验研究

An experimental study on the effect of capsaicin on hepatic stellate cells and liver fibrogenesis

目的 探讨辣椒素对大鼠肝星状细胞（hepatic stellate cells,HSCs）活性的影响及其对活体肝纤维化的治疗作用.方法 DCFH-DA法检测辣椒素对HSCs中活性氧的影响；用酶标法检测辣椒素对HSCs增殖的影响；Western blot检测HSCs α-肌动蛋白（α-SMA）的表达；RT-PCR检测辣椒素对HSCs纤维化相关基因表达的影响；流式细胞仪检测辣椒素对HSCs凋亡的影响；Western blot法检测bcl-2、bax、细胞色素酶c（cytochrome,cyt-c）等蛋白的表达；建立鼠肝纤维化模型,经腹腔输注辣椒素,检测肝组织病理切片,肝纤维化指标.结果 辣椒素抑制HSCs中活性氧的产生,抑制HSCs的活化（F=57.392,P＜0.05）与增殖（F=13.267,P＜0.05）,促进HSCs的凋亡（F=235.571,P＜0.05）,促进凋亡相关蛋白Bax、cyt-c、caspase-3的表达（分别F=29.334,38.274,138.329,均P＜0.05）.改变HSCs纤维化相关基因TGF-β1、TIMP-1的表达（分别F=376.534,253.751,均P＜0.05）,活体实验显示辣椒素可降低肝羟脯氨酸含量及血清Ⅲ型胶原和透明质酸水平（分别F=153.397,27.149,38.392,均P＜0.05）.结论 辣椒素抑制HSCs增殖活化并诱导其凋亡.辣椒素对肝纤维化具有一定的抑制作用.

Objective To investigate the effect of capsaicin on hepatic stellate cells （HSCs） and liver fibrogenesis.Methods HSCs were cultured.The reactive oxygen in HSCs under capsaicin at different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was detected by CCK-8 test kit.Smoothmuscle α-actin （α-SMA） expression of HSCs was evaluated by Western blot.The fibrosisrelated genes were tested by RT-PCR.The apoptosis of HSCs was measured by flow cytometer.Bcl-2,bax and cyt-c was detected by Western blot.A murine model of liver fibrogenes was established.Capsaicin of different concentration was injected intraperitoneally.Liver pathology was observed using HE staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results In dose dependent manner capsaicin inhibited the generation of the reactive oxygen species.Proliferation and activation of HSCs was inhibited by capsaicin （respectively F =13.267,57.392,all P 〈 0.05） and the apoptosis of HSCs was promoted by capsaicin （F =235.571,P 〈 0.05）.Bax,cyt-c and caspase-3 was increased obviously （respectively F =29.334,38.274,138.329,all P 〈 0.05）.Capsaicin changed the expression of fibrosis-related genes （TGF-β1,TIMP-1） in HSCs （respectively F =376.534,253.751,all P 〈0.05）.Capsaicin downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid in the rat model （respectively F =153.397,27.149,38.392,all P 〈 0.05）.Conclusions Capsaicin inhibits the proliferation and activation of hepatic stellate cells.Capsaicin promotes the apoptosis of hepatic stellate cells,and inhibits liver fibrogenesis.