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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (2): 171-175.doi: 10.12047/j.cjap.5956.2020.038

• 研究论文 • 上一篇    下一篇

白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响*

杜洋1, 邵淑丽1,2△, 焦凯贺1, 刘祥露1, 冯元1, 张伟伟1,2   

  1. 1.齐齐哈尔大学生命科学与农林学院, 2.抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江 齐齐哈尔 161006
  • 出版日期:2020-03-28 发布日期:2020-07-31
  • 通讯作者: Tel: 13836267458; E-mail: shshl32@163.com
  • 基金资助:
    *黑龙江省自然科学基金(C201241);黑龙江省教育厅基本业务专项重点项目(135109104);黑龙江省高教强省优势特色学科—玉米“粮头食尾”重点专项(LTSW201737);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201809)

Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45

DU Yang1, SHAO Shu-li1,2△, JIAO Kai-he1, LIU Xiang-lu1, FENG Yuan1, ZHANG Wei-wei1,2   

  1. 1. College of Life Sciences, Agriculture and Forestry; 2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihar 161006, China
  • Online:2020-03-28 Published:2020-07-31

摘要: 目的: 探讨白花蛇舌草(注射液)对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响。方法: 将人胃癌细胞MNK-45分成4组,每组设置3个复孔,对照组为未加入白花蛇舌草的MNK-45细胞,3组实验组分别加入终浓度为20、30、40 μg/ml白花蛇舌草的MNK-45细胞,各组在5%的CO2培养箱中孵育48 h后,利用激光共聚焦显微镜下观察细胞形态变化,流式细胞术检测线粒体膜电位,qRT-PCR检测Cytochrome C (Cyt c)、caspase3caspase9基因的表达变化,Western blot检测Cytochrome C (Cyt c)、caspase3和caspase9蛋白的表达变化。结果: 与对照组相比,终浓度为20、30、40 μg/ml的各白花蛇舌草处理组,其MNK-45细胞的线粒体膜电位均明显降低(P<0.01),Cyt ccaspase3caspase9的基因表达均明显上调(P<0.01)、蛋白表达也均显著升高(P<0.05或 P<0.01),40 μg/ml的白花蛇舌草处理组的表现最佳。结论: 在终浓度为20~40 μg/ml的范围内,白花蛇舌草能够降低人胃癌MNK-45细胞线粒体膜电位, 诱导细胞凋亡,并可上调Cyt c、caspase3caspase9基因表达。

关键词: 白花蛇舌草, 线粒体膜电位, 人胃癌MNK-45细胞, 细胞色素C, caspase3, caspase9

Abstract: Objective: To investigate the effects of hedyotis diffusa (injection) on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45 cells. Methods: The human gastric cancer MNK-45 cells were divided into 4 groups, each group was set with 3 replicates. The control group was MNK-45 cells without added hedyotis diffusa; the 3 groups of experimental groups were treated with hedyotis diffusa at final concentrations of 20 , 30, 40 μg / ml respectively; each group was incubated for 48 h in a 5% carbon dioxide incubator, and the morphological changes of the cells were observed under a laser confocal microscope. Mitochondrial membrane potential was detected by flow cytometry. The expressions of Cytochrome C (Cyt c), caspase3 and caspase9 genes and proteins were detected by qRT-PCR and Western blot respectively. Results: Compared with the control group, the mitochondrial membrane potentials of MNK-45 cells were significantly reduced in the hedyotis diffusa treated groups at final concentrations of 20, 30, and 40 μg / ml (P<0. 01). The gene expressions of Cyt c, caspase3, and caspase9 were significantly up-regulated (P<0. 01) and their protein expressions were also significantly increased (P<0. 05 or P<0. 01). The 40 μg / ml hedyotis diffusa treatment group performed best. Conclusion: In the final concentration range of 20 ~ 40 μg / ml, hedyotis diffusa can reduce human gastric cancer MNK-45 cells mitochondrial membrane potential, induce apoptosis and up-regulate Cyt c, caspase3 and caspase9 gene expressions.

Key words: hedyotis diffusa, mitochondrial membrane potential, human gastric cancer MNK-45 cells, Cyt c, caspase3, caspase9

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