天麻素调控PINK1/Parkin-VDAC1信号改善CoCl2诱导的HT22细胞凋亡  

Inhibitory effect of gastrodin on HT22 apoptosis induced by CoCl2 via PINK1/Parkin-VDAC1 signal

作　　者：冉芮 陈婷婷 陈永鑫 余俊杰 李悦[2] 甘诗泉 陶玲[1] 沈祥春[1] RAN Rui;CHEN Tingting;CHEN Yongxin;YU Junjie;LI Yue;GAN Shiquan;TAO Ling;SHEN Xiangchun(Key Laboratory of Utilization of Natural Medicine Resources,Guizhou Medical University,Guiyang 550025,China;Department of Pharmacy,Guiyang Maternal and Child Health Care Hospital,Guiyang 550003,China)

机构地区：[1]贵州医科大学天然药物优效利用重点实验室,贵阳550025 [2]贵阳市妇幼保健院药学部,贵阳550003

出　　处：《中华中医药杂志》2023年第7期3109-3114,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82060772);贵州医科大学国家自然科学基金培育项目(No.19NSP07);贵阳市卫生健康局科学技术计划项目(No.[2022]016);贵州省卫生健康委科学技术基金项目(No.Gzwkj2021-465)。

摘　　要：目的:探讨天麻素对二氯化钴(CoCl2)诱导的HT22细胞凋亡作用机制。方法:体外建立CoCl2损伤HT22的细胞模型。分组为空白组,模型组,天麻素低、中、高剂量组,Annexin V-FITC/PI双染法检测细胞凋亡率、JC-1检测线粒体膜电位(MMP)、DCFH-DA检测活性氧(ROS)水平、二硫代硝基苯甲酸(DTNB)检测谷胱甘肽(GSH)含量;VDAC1基因过表达以及使用Parkin活性抑制剂后,Western Blot检测凋亡相关蛋白表达水平。结果:天麻素降低HT22细胞凋亡率,抑制HT22细胞MMP降低,抑制HT22细胞ROS水平升高和GSH含量降低。Western Blot显示,天麻素抑制PINK1、Parkin和VDAC1表达,同时也能降低凋亡相关蛋白(Bax/Bcl-2比率、Cleaved-Caspase 3、Caspase 3、Cytochrome-c)表达水平;VDAC1过表达以及使用Parkin活性抑制剂Suramin后,PINK1与Parkin的蛋白表达水平不受影响,天麻素仍然表现出抑制凋亡相关蛋白水平的作用。结论:天麻素可能通过抑制PINK1/Parkin-VDAC1信号下调HT22细胞凋亡水平。Objective:To investigate the mechanism of gastrodin on apoptosis induced by CoCl2 in HT22 cells.Methods:CoCl2 was used to act on HT22 cells to establish neuron apoptosis model in vitro.HT22 cells were divided into blank control group,model control group,low dose gastrodin group,middle dose gastrodin group and high dose gastrodin group.Annexin V-FITC/PI double staining was used to detect apoptosis rate,JC-1 to detect mitochondrial membrane potential(MMP),DCFH-DA to detect reactive oxygen species(ROS)level and DTNB to detect glutathione(GSH)content.After VDAC1 Overexpressed and Parkin activity inhibitor used,the expression level of protein was detected by Western Blot.Results:Gastrodin reduced the apoptosis rate,reverse the decreased of MMP,inhibited the increase of ROS level,and rose the content of GSH in HT22 cells induced by CoCl2.Gastrodin aslo inhibited the expression of PINK1,Parkin and VDAC1,and decreased the expression level of apoptosis-related proteins(Bax/Bcl-2 ratio,Cleaved-Caspase 3,Caspase 3 and Cytochrome-c),showed by Western Blot Gastrodin could still inhibit the level of apoptosis-related proteins,while the protein expression levels of PINK1 and Parkin were not affected,after VDAC1 overexpressed and Parkin activity inhibitor used.Conclusion:Gastrodin may down-regulate HT22 cells apoptosis by inhibiting Pink1/Parkin-VDAC1 signal.

关 键 词：天麻素 PINK1/Parkin 电压依赖性阴离子通道蛋白1 HT22细胞 细胞凋亡 

分 类 号：R285[医药卫生—中药学]