枫杨提取物对MH7A细胞增殖和凋亡的影响及其机制研究

Effects and mechanism of Pterocarya Hupehensis Skan extracts on proliferation and apoptosis of MH7A cells

目的:观察枫杨不同提取物对人类风湿关节炎成纤维滑膜细胞MH7A增殖和凋亡的影响,初步探讨其诱导MH7A细胞凋亡的可能机制。方法:制备枫杨乙醇、石油醚、氯仿、乙酸乙酯和正丁醇提取物后分别作用于MH7A细胞,检测其增殖情况。取增殖抑制作用最好的提取物作用于MH7A细胞,检测其凋亡情况,Caspase-3和Caspase-9的酶活性; Cyt-C、Cleaved Caspase-3、Cleaved Caspase-9、Bcl-2和BAX的蛋白表达,P53、BAX、BAK、Bcl-2和Bcl-xL的mRNA表达。结果:与枫杨石油醚、氯仿、乙酸乙酯和正丁醇提取物相比,枫杨乙醇提取物可明显抑制MH7A细胞增殖,诱导其凋亡,使细胞内Caspase-3和Caspase-9的酶活性升高,Cyt-C、Cleaved Caspase-3、Cleaved Caspase-9和BAX的蛋白表达升高,Bcl-2的蛋白表达下降; P53、BAX、BAK的mRNA表达升高,Bcl-2、Bcl-xL的mRNA表达降低。结论:枫杨乙醇提取物可明显抑制MH7A细胞增殖,诱导其凋亡,并调控Cyt-C、P53、Caspase家族、Bcl-2家族相关蛋白的表达。因此推测该提取物可能通过影响线粒体凋亡途径诱导人类风湿关节炎成纤维滑膜细胞MH7A的凋亡,抑制其增殖。

Objective: To observe Pterocarya Hupehensis Skan different extracts on proliferation and apoptosis of human rheumatoid arthritis fibroblast-like synovial MH7 A cells,and then to explore the possible mechanism of induced apoptosis of MH7 A cells. Methods: Prepared different extracts of Pterocarya Hupehensis Skan with ethanol,petroleum ether,chloroform,ethyl acetate,and nbutanol,and then to co-cultured MH7 A cells with each extract,detected the proliferation of MH7 A cells. The MH7 A cells were treated with extracts of the active site with the best inhibitory effect on proliferation,and then to detect their apoptosis. Caspase-3 and Caspase-9 enzyme activity. Cyt-C,Cleaved Caspase-3,Cleaved Caspase-9,Bcl-2 and BAX protein expression. P53,BAX,BAK,Bcl-2 and Bcl-xL mRNA expression. Results: Compared with extracts of Pterocarya Hupehensis Skan with petroleum ether,chloroform,ethyl acetate and nbutanol,ethanol extract could obviously inhibit the proliferation of MH7 A cells and induce apoptosis of it. Ethanol extract made the enzyme activity of Caspase-3 and Caspase-9 in the cells increased. The protein expression of Cyt-C,Cleaved Caspase-3,Cleaved Caspase-9 and BAX increased,and the protein expression of Bcl-2 decreased. The mRNA expression of P53,BAX and BAK increased,and the mRNA expression of Bcl-2 and Bcl-xL decreased simultaneously. Conclusion: The ethanol extract of Pterocarya Hupehensis Skan could obviously inhibit the proliferation of MH7 A cells and induce apoptosis of it,and regulated and controled the protein expression of Cyt-C,P53,Caspase family and Bcl-2 family. These results sugest that ethanol extract of Pterocarya Hupehensis Skan could induce the apoptosis of human rheumatoid arthritis fibroblast-like synovial MH7 A cell by activating the mitochondrial apoptotic pathways.