Zhao Jia-Rong, Feng Xue-Song, Li Hong, Wang Guo-Quan, Zhang Yan-Fang, Shi Hai-Long, Li Yuan-Yuan, Chao Xu
The College of Basic Medicine Sciences, Shaanxi University of Chinese Medicine, Xianyang 712046.
The Second Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712000.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Sep;36(5):489-493. doi: 10.12047/j.cjap.5839.2020.104.
To investigate the effects of Sini San prescription(SNS) on the proliferation and apoptosis of HepG2 cells and its molecular mechanism. The morphological changes of hepatocellular carcinoma HepG2 cells treated by SNS were observed by inverted microscope. MTT assay was used to detect the inhibitory effect of SNS on cell proliferation. Fluorescence staining and flow cytometry were employed to analyze the effect of SNS on apoptosis of HepG2 cells. Rho123 (Rhodamine 123) staining method was performed to detect the changes of mitochondrial membrane potential, and Western blot was used to evaluate the expression of proteins related to apoptosis. The number of hepatocellular carcinoma HepG2 cells were significantly decreased (＜0.01) and cells showed typical apoptotic cell morphology after treated with serum contained SNS. The inhibition rate of HepG2 cells was increased with the increase of concentration of serum contained SNS. The number of cells in G1 phase was significantly increased, while G2 phase was decreased after treated with serum contained SNS(＜0.05).The apoptosis rate and mitochondrial membrane potential of HepG2 cells were significantly increased and decreased after treated with serum contained SNS(＜0.05). The expression levels of Bax, caspase-3,-9 and cyt-c were significantly increased, while the expression of bcl-2 was decreased in HepG2 cells treated with serum contained SNS(＜0.05). Sini San prescription can inhibit the proliferation of HepG2 cells and induce apoptosis by mitochondrial pathway.
探讨四逆散方剂（SNS）对肝癌HepG2细胞增殖和凋亡的影响及其分子机制。通过倒置显微镜观察SNS处理的肝癌HepG2细胞的形态变化。采用MTT法检测SNS对细胞增殖的抑制作用。利用荧光染色和流式细胞术分析SNS对HepG2细胞凋亡的影响。采用罗丹明123（Rho123）染色法检测线粒体膜电位的变化，并用蛋白质免疫印迹法评估凋亡相关蛋白的表达。含SNS血清处理后，肝癌HepG2细胞数量显著减少（＜0.01），细胞呈现典型的凋亡细胞形态。含SNS血清处理后，HepG2细胞的抑制率随血清浓度的增加而升高。含SNS血清处理后，G1期细胞数量显著增加，而G2期细胞数量减少（＜0.05）。含SNS血清处理后，HepG2细胞的凋亡率显著升高，线粒体膜电位显著降低（＜0.05）。含SNS血清处理的HepG2细胞中，Bax、caspase-3、-9和细胞色素c的表达水平显著升高，而bcl-2的表达降低（＜0.05）。四逆散方剂可抑制HepG2细胞的增殖，并通过线粒体途径诱导细胞凋亡。
