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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (5): 489-493.doi: 10.12047/j.cjap.5839.2020.104

• 研究论文 • 上一篇    下一篇

四逆散对人肝癌HepG2细胞增殖、凋亡的影响及其机制*

赵家荣1, 冯雪松1, 李宏2, 王国全3, 张艳芳1, 史海龙1, 李媛媛2, 晁旭1,2△   

  1. 1.陕西中医药大学基础医学院, 咸阳 712046;
    2.陕西中医药大学第二附属医院, 咸阳 712000;
    3.陕西中医药大学药学院, 咸阳 712000
  • 收稿日期:2019-03-05 修回日期:2020-07-20 发布日期:2021-02-25
  • 通讯作者: Tel: 029-33574065; E-mail: chaoxu2004@126.com
  • 基金资助:
    *国家自然科学基金(81774132);陕西省中医药管理局科研项目(2019-zz-JC012);陕西省中医药大学学科创新团队项目(2019-YS05);陕西中医药大学第二附属医院学科创新团队项目(2020XKTD-A01)

Effect of Sini San prescription on apoptosis and proliferation of hepatocellular carcinoma HepG2 cells

ZHAO Jia-rong1, FENG Xue-song1, LI Hong2, WANG Guo-quan3, ZHANG Yan-fang1, SHI Hai-long1, LI Yuan-yuan2, CHAO Xu1,2△   

  1. 1. The College of Basic Medicine Sciences, Shaanxi University of Chinese Medicine, Xianyang 712046;
    2.The Second Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712000;
    3. College of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang 712046, China
  • Received:2019-03-05 Revised:2020-07-20 Published:2021-02-25

摘要: 目的: 探讨含四逆散药液血清对人肝癌HepG2细胞增殖、凋亡的影响及机制。方法: 将人肝癌HepG2细胞分为5组,每组3个复孔。实验组细胞用五氟尿嘧啶(5-FU)或不同浓度的含四逆散药液血清处理48 h后,用倒置显微镜观察含四逆散药液血清处理后人肝癌HepG2细胞形态的变化;MTT法检测含四逆散药液血清对HepG2细胞生长的抑制作用;荧光染色和流式细胞术分别分析含四逆散药液血清对HepG2细胞凋亡的影响。Rho123染色法检测线粒体膜电位变化,Western blot检测细胞凋亡相关蛋白的表达。结果: 与对照组比较,含四逆散药液血清处理人肝癌HepG2细胞后,细胞数量显著减少(P<0.01),形态发生改变,呈现典型的凋亡细胞形态;G1期细胞数明显增加,而G2 期细胞数量显著减少(P<0.05);Bax、Caspase-3、-9和Cyt-c的表达显著升高,而Bcl-2的表达显著降低(P<0.05);随着含四逆散药液血清浓度增大,HepG2细胞线粒体膜电位显著下降(P<0.05)。结论: 四逆散可以抑制HepG2细胞增殖,并通过线粒体途径诱导细胞凋亡。

关键词: 四逆散, 肝癌, HepG2细胞, 凋亡

Abstract: Objective: To investigate the effects of Sini San prescription(SNS) on the proliferation and apoptosis of HepG2 cells and its molecular mechanism. Methods: 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. Results: The number of hepatocellular carcinoma HepG2 cells were significantly decreased (P<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(P<0.05).The apoptosis rate and mitochondrial membrane potential of HepG2 cells were significantly increased and decreased after treated with serum contained SNS(P<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(P<0.05).Conclusion: Sini San prescription can inhibit the proliferation of HepG2 cells and induce apoptosis by mitochondrial pathway.

Key words: Sini San prescription, hepatocellular carcinoma, HepG2 cells, apoptosis

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