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、40μg/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)、caspase3和caspase9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、40μg/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 c、caspase3和caspase9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),40μg/ml的白花蛇舌草处理组的表现最佳。结论:在终浓度为20~40μg/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、caspase3和caspase9基因表达。
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 20,30,40μg/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 40μg/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 40μg/ml hedyotis diffusa treatment group performed best.Conclusion:In the final concentration range of 20~40μg/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.
作者
杜洋
邵淑丽
焦凯贺
刘祥露
冯元
张伟伟
DU Yang;SHAO Shu-li;JIAO Kai-he;LIU Xiang-lu;FENG Yuan;ZHANG Wei-wei(College of Life Sciences,Agriculture and Forestry;Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihar 161006,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2020年第2期171-175,共5页
Chinese Journal of Applied Physiology
基金
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)
