人参皂苷Rg1对氧糖剥夺/复氧复糖损伤PC12细胞的保护机制  

Ginsenoside Rg1 protects against oxygen-glucose deprivation/reperfusion injury in PC12 cells

作　　者：叶劲涛 李锋涛[1] 宋焕瑾[1] 薛建利[1] 林磊[2] 程斌[1] Ye Jintao;Li Fengtao;Song Huanjin;Xue Jianli;Lin Lei;Cheng Bin(Department of Orthopedics, Second Affiliated Hospital of Xi'an Jiaotong University Medical School, Xi'an 710004, Shaanxi Province, China;Department of Orthopedics, Hanzhong Central University, Hanzhong 723000, Shaanxi Province, China)

机构地区：[1]西安交通大学医学院第二附属医院骨科,陕西省西安市710004 [2]汉中市中心医院骨科,陕西省汉中市723000

出　　处：《中国组织工程研究》2019年第7期1090-1096,共7页Chinese Journal of Tissue Engineering Research

基　　金：陕西省自然科学基金面上项目(2014JM2_8157);项目负责人:程斌;陕西省自然科学基金面上项目(2016JM8129);项目负责人:宋焕瑾~~

摘　　要：背景:人参皂苷具有抗肿瘤、抗氧化、抗疲劳、抗衰老、降血脂、增强记忆力、提高免疫力等作用,其中Rb1和Rg1是人参皂苷的最主要的活性成分。但人参皂苷Rg1对氧糖剥夺诱导的PC12细胞凋亡是否也有保护作用,尚不清楚。目的:观察人参皂苷Rg1预处理对于PC12细胞氧糖剥夺/复氧复糖模型中PC12细胞活力、survivin及caspase-3表达和细胞凋亡的影响。方法:采用构建的PC12细胞氧糖剥夺/复氧复糖模型,并使用5,10,20,40μmol/L的人参皂苷Rg1对其进行预处理,以正常细胞做对照。通过MTT法检测细胞活性,通过免疫细胞化学技术检测survivin及caspase-3蛋白,通过TUNEL法检测细胞凋亡。结果与结论:(1)细胞活力检测氧糖剥夺/复氧复糖组细胞活性明显下降(P <0.05),人参皂苷Rg1干预后药物组各亚组细胞活性均较氧糖剥夺/复氧复糖组有所恢复,但仍较正常组为低(P <0.05);(2)人参皂苷Rg1干预使survivin阳性细胞增多,caspase-3阳性细胞和TUNEL阳性细胞计数值减少;(3)神经元凋亡出现与人参皂苷Rg1诱导的survivin成负相关,与caspase-3成正相关。(4)结果说明,人参皂苷Rg1对于PC12细胞氧糖剥夺/复氧复糖处理后的survivin蛋白表达具有促进作用,并通过促进survivin蛋白的表达来对抗caspase-3的表达,从而抑制细胞凋亡,且该作用具有剂量依赖关系。BACKGROUND: Ginsenoside has anti-tumor, anti-oxidative, anti-fatigue, anti-aging, hypolipidemic, memory enhancement, immunity enhancement effects, and Rb1 and Rg1 are the most important active components of ginsenosides. However, it is unclear whether ginsenoside Rg1 also protects against apoptosis in PC12 cells induced by oxygen-glucose deprivation (OGD). OBJECTIVE: To investigate the effects of pretreatment with ginsenoside Rg1 on PC12 cell viability, survivin and caspase-3 expression and apoptosis in PC12 cells induced by OGD/reperfusion. METHODS: In the OGD/reperfusion model, the PC12 cells were pretreated with 5, 10, 20, 40 μmol/L ginsenoside Rg1. Normal cells were used as controls. Cell viability was determined by MTT assay. Expression of survivin and caspase-3 proteins was measured by immunocytochemistry. Apoptosis in PC12 cells was detected by TUNEL assay. RESULTS AND CONCLUSION:(1) The cell viability was significantly decreased after OGD/reperfusion (P < 0.05). Pretreatment with ginsenoside Rg1 could elevate the cell viability, but it was still lower than the normal level (P < 0.05).(2) Pretreatment with ginsenoside Rg1 increased survivin positive cells, but decreased caspase-3 positive cells and TUNEL positive cells.(3) Neuronal apoptosis was negatively correlated with the ginsenoside Rg1-induced survivin, but positively correlated with the caspase-3. All these findings indicate that ginsenoside Rg1 pretreatment can promote the expression of survivin in PC12 cells after OGD/reperfusion, and moreover, this promotion effect on survivin can further inhibit the expression of caspase-3, thereby suppression apoptosis in PC12 cells, in a dose-dependent manner.

关 键 词：PC12细胞 人参皂苷RG1 SURVIVIN蛋白 caspase-3 组织构建 人参皂甙类 半胱氨酸天冬氨酸蛋白酶3 组织工程 

分 类 号：R446[医药卫生—诊断学] R318[医药卫生—临床医学]