摘要
目的:探讨黄芪甲苷(AS-Ⅳ)对小鼠睾丸间质(TM3)细胞具有抗氧化应激并抑制凋亡的作用。方法:用不同浓度(0、100、200、300μmol/L)的过氧化氢(H_(2)O_(2))分别处理TM3细胞12、24、48 h,确定建立氧化损伤细胞模型的适宜浓度为100μmol/L作用48 h;将TM3细胞分为空白组(正常培养液),模型组(100μmol/L H_(2)O_(2)),10、50、100 mg/L AS-Ⅳ组分别培养48 h后,通过CCK-8检测细胞增殖能力,流式细胞术检测细胞凋亡,JC-1染色法检测细胞中线粒体膜电位,探针法检测细胞活性氧(ROS)水平,Western Blot检测Caspase9、Bax蛋白表达水平。结果:AS-Ⅳ可以降低Caspase9、Bax蛋白表达水平,抑制细胞内线粒体膜电位下降,减少细胞内ROS的生成,降低TM3细胞凋亡率。结论:AS-Ⅳ可以减轻H_(2)O_(2)诱导的小鼠TM3细胞氧化应激反应并抑制细胞凋亡。
Objective:To investigate the effects of AstragalosideⅣ(AS-Ⅳ)on the anti-oxidative stress and anti-apoptosis in TM3 mouse Leydig cells.Methods:TM3 Leydig cells were treated with different concentrations(0,100,200,300μmol/L)of hydrogen peroxide(H_(2)O_(2))for 12 h,24 h,48 h,and the appropriate concentration was 100μmol/L for 48 h.TM3 Leydig cells were divided into blank group(normal culture medium),model group(100μmol/L H_(2)O_(2)),10 mg/L AS-Ⅳgroup,50 mg/L AS-Ⅳgroup and 100 mg/L AS-Ⅳgroup for 48 h,respectively.The cell proliferation capacity was detected by CCK-8.Cell apoptosis was detected by flow cytometry,mitochondrial membrane potential was detected by JC-1 staining,reactive oxygen species(ROS)level was detected by probe method,and protein expression levels of Caspase9 and Bax were detected by Western Blot.Results:AS-Ⅳtreatment reduced the protein expression levels of Caspase9 and Bax,inhibited the decrease of mitochondrial membrane potential,reduced the generation of intracellular ROS,and reduced the apoptosis rate of TM3 leydig cells.Conclusion:AS-Ⅳreduces H_(2)O_(2) induced oxidative stress and apoptosis in TM3 Leydig cells.
作者
李泓莹
林煦垚
史孟华
杨艳芳
马天宇
胡召娣
黄颖
徐磊
陈雪松
王绍年
秦国政
LI Hong-ying;LIN Xu-yao;SHI Meng-hua;YANG Yan-fang;MA Tian-yu;HU Zhao-di;HUANG Ying;XU Lei;CHEN Xue-song;WANG Shao-nian;QIN Guo-zheng(First Clinical College of Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China;Yunnan Provincial Hospital of Traditional Chinese Medicine/The First Affiliated Hospital of Yunnan University of Chinese Medicine,Kunming 650021,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第5期2456-2459,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
云南省教育厅科学研究基金项目(No.2022Y331)。
