摘要
目的探讨黄芪甲苷对PC12细胞氧化应激损伤的作用。方法体外培养PC12细胞,6-羟基多巴胺(6-OHDA)作用PC12细胞导致氧化应激损伤。根据细胞作用方法随机分为4组:①对照组,采用完全培养基正常培养;②6-OHDA组,给予终浓度为100μmol/L的6-OHDA处理24 h;③低、中、高剂量黄芪甲苷组,分别给予终浓度为25、50、100μmol/L的黄芪甲苷预处理24 h,然后给予终浓度为100μmol/L 6-OHDA处理24 h;④AG490组,以20μmol/L AG490(JAK2/STAT3信号通路抑制剂)预处理16 h,加入终浓度为100μmol/L黄芪甲苷处理24 h,然后再给予终浓度为100μmol/L 6-OHDA处理24 h。CCK8法检测细胞生存率,流式细胞仪检测细胞凋亡率,酶联免疫吸附实验法检测细胞上清液超氧化物歧化酶(SOD)和丙二醛(MDA)水平,免疫印迹法检测细胞p-JAK2和p-STAT3蛋白表达。结果6-OHDA作用后,PC12细胞存活率明显降低,细胞凋亡率明显升高,细胞培养液SOD水平明显降低、MDA水平明显升高,细胞p-JAK2和p-STAT3蛋白表达水平明显降低;黄芪甲苷预处理明显逆转6-OH⁃DA的作用,而且呈剂量依赖性;AG490预处理明显逆转黄芪甲苷的作用。结论6-OHDA作用PC12细胞,可导致氧化应激损伤促使细胞凋亡;黄芪甲苷预处理能激活JAK2/STAT3信号通路,抑制6-OHDA对PC12细胞的损伤,对PC12细胞起保护作用。
Objective To investigate the effect of astragiosideⅣ(ASⅣ)on the 6-hydroxydopamine(6-OHDA)-induced oxidative stress injury in PC12 cells.Methods PC12 cells were cultured in vitro and the 6-hydroxy dicamine(6-OHDA)was used to cause oxidative stress injury in the PC12 cells.The PC12 cells were randomly divided into 4 groups:①control group,the PC12 cells were cultured in medium without any drugs;②6-OHDA group,the PC12 cells were treated by 6-OHDA for 24 h at a final concentration of 100μmol/L;③low,medium,and high dose ASⅣgroups,the PC12 cells were treated by ASⅣfor 24 h at a final concentration of 25,50,100μmol/L,respectively,and then treated by 6-OHDA for 24 h;④Ag490 group,the PC12 cells were trated by AG490(JAK2/STAT3 signaling inhibitor)for 16 h at a final concentration of 20μmol/L,then treated by ASⅣfor 24 h at a final concentration of 100μmol/L,and finally treated by 6-OHDA for 24 h.CCK8 method was used to detect the cell survival rate.Cytometry was used to detect the cell apoptosis rate.Enzyme-linked immunosorbent assay was used to detect the cell supernatant levels of superoxide dismutase(SOD)and malondialdehyde(MDA).Western blotting was used to detect the protein expression levels of p-JAK2 and p-STAT3.Results After 6-OHDA treatment,the survival rate of PC12 cells significantly reduced,and the apoptosis rate significantly increased,the SOD level in cell culture medium significantly reduced,the MDA level significantly increased,and the expression levels of p-JAK2 and p-STAT3 protein significantly reduced.ASⅣpretreatment significantly reversed the effect of 6-OHDA on the PC12 cellsdose-dependent manner.AG490 pretreatment significantly reversed the effect of ASⅣ.Conclusions 6-OHDA treatment can result in oxidative stress injury in PC12 cells and then cause cell apoptosis in PC12 cells.ASⅣpretreatment can significantly inhibit the 6-OHDA-induced damage to PC12 cells by activating JAK2/STAT3 signaling pathways.
作者
黄万刚
杨东风
冯佳良
黄淮
高超
徐正虎
HUANG Wan-gang;YANG Dong-feng;FENG Jia-liang;HUANG Huai;GAO Chao;XU Zheng-hu(Department of Neurosurgery,Hebei Petro China Central Hospital,Langfang 065000,China;Department of EmergencyHebei Petro China Central Hospital,Langfang 065000,China;Department of Neurology,Hebei Petro China Central Hospital,Langfang 065000,China)
出处
《中国临床神经外科杂志》
2021年第4期270-273,共4页
Chinese Journal of Clinical Neurosurgery
基金
廊坊市科技支撑计划项目(2020013092)。
