实时动态法分析七味红花殊胜丸抑制肝癌细胞线粒体代谢

Real-time dynamic analysis of Qiwei Honghua Shusheng Pill inhibiting mitochondria metabolism of liver cancer cells

目的:观察七味红花殊胜丸对人肝癌SMMC-7721细胞线粒体代谢酶的影响。方法:运用实时动态法分析七味红花殊胜丸对细胞活性的影响及各给药组不同时间IC50值的变化情况。采用MTT法检测七味红花殊胜丸对SMMC-7721细胞的抑制率;ELISA法检测七味红花殊胜丸对SMMC-7721细胞线粒体代谢和凋亡的影响,Western Blot法检测凋亡蛋白的表达状况。结果:实时动态检测结果提示,七味红花殊胜丸5 mg·mL^(-1)作用48 h对SMMC-7721细胞的抑制率最高(P<0.01);ELISA检测结果提示,七味红花殊胜丸能降低SMMC-7721细胞内ATP、ATPase、线粒体呼吸链复合物Ⅰ/Ⅱ/Ⅲ/Ⅳ酶的表达量,影响细胞的能量代谢;升高促凋亡蛋白Bax、Caspase-3和Caspase-9的表达量,降低抗凋亡蛋白Bcl-2的表达量,诱导细胞凋亡。结论:七味红花殊胜丸可通过影响细胞的能量代谢,诱导SMMC-7721细胞凋亡。

OBJECTIVE To observe the effect of Qiwei Honghua Shusheng Pill on SMMC-7721 hepatoma cell mitochondria.METHODS The effects of each dosing group on cellular activity and IC50 value of each timepoint were analyzed by real-time dynamic detection.Methyl thiazolyl tetrazolium(MTT)colorimetry was employed for observing the inhibition rate of Qiwei Honghua Shusheng Pill on human liver cancer SMMC-7721 cell and enzyme-linked immunosorbent assay(ELISA)for detecting the effect of Qiwei Honghua Shusheng Pill on energy metabolism and apoptosis.The expression of apoptotic protein was detected by Western blot.RESULTS The results of real-time dynamic detection indicated that 48 h was the best timepoint and 5 mg·mL^(-1) the best concentration.MTT implied that Qiwei Honghua Shusheng Pill could inhibit cellular proliferation after 48 h in a concentration-dependent manner(P<0.01).ELISA showed that Qiwei Honghuashusheng Pill with large/medium doses(10/5 mg·mL^(-1))could lower the expressions of ATP,ATPase and mitochondrial respiratory chain complexⅠ/Ⅱ/Ⅲ/Ⅳenzymes and affect cellular energy metabolism.It could also enhance the contents of pro-apoptotic proteins Bax,caspase-3 and caspase-9,down-regulate the expression of anti-apoptotic protein Bcl-2 and induce the apoptosis of liver cancer cells.CONCLUSION Qiwei Honghua Shusheng Pill may induce the apoptosis of liver cancer cells through regulating cellular energy metabolism.