黄芪甲苷改善1-甲基-4-苯基吡啶离子诱导的帕金森病SK-N-SH细胞损伤疗效初探  

Effect of astragalosideⅣon SK-N-SH cells damage induced by 1-methyl-4-phenylpyridine in Parkinson’s disease

作　　者：黄晓静 黄淮 徐正虎 黄万钢 杨东风 任贺成[4] HUANG Xiao-jing;HUANG Huai;XU Zheng-hu;HUANG Wan-gang;YANG Dong-feng;REN He-cheng(Department of Neurology,Hebei Petro China Central Hospital,Langfang 065000,Hebei,China;Department of Neurosurgery,Hebei Petro China Central Hospital,Langfang 065000,Hebei,China;Department of Emergency,Hebei Petro China Central Hospital,Langfang 065000,Hebei,China;Department of Neurosurgery,Tianjin Huanhu Hospital,Tianjin 300350,China)

机构地区：[1]河北中石油中心医院神经内科,廊坊065000 [2]河北中石油中心医院神经外科,廊坊065000 [3]河北中石油中心医院急诊科,廊坊065000 [4]天津市环湖医院神经外科,300350

出　　处：《中国现代神经疾病杂志》2022年第3期155-162,共8页Chinese Journal of Contemporary Neurology and Neurosurgery

基　　金：河北省廊坊市2020年科技支撑计划项目(项目编号:2020013092)。

摘　　要：目的探讨黄芪甲苷对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病SK-N-SH细胞损伤的作用机制。方法人神经母细胞瘤细胞系SK-N-SH培养至对数生长期,随机予以常规培养(对照组)、MPP+诱导(MPP+组)、黄芪甲苷10 mg/ml+MPP+诱导(黄芪甲苷10 mg/ml组)、黄芪甲苷30 mg/ml+MPP+诱导(黄芪甲苷30 mg/ml组)、黄芪甲苷30 mg/ml+MPP+诱导+Janus激酶2(JAK2)抑制剂AG490(JAK2抑制剂组),噻唑蓝法检测细胞存活率,流式细胞术检测细胞凋亡率,DCFH-DA荧光探针测定活性氧(ROS)含量,酶法测定乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)活性,Western blotting法测定磷酸化JAK2(pJAK2)和磷酸化信号转导与转录激活因子3(pSTAT3)蛋白相对表达量。结果不同处理组SK-N-SH细胞存活率(P=0.000)、凋亡率(P=0.000)、ROS含量(P=0.000)、LDH活性(P=0.000)、SOD活性(P=0.003)、pJAK2(P=0.000)和pSTAT3(P=0.000)蛋白相对表达量差异有统计学意义。与对照组相比,MPP+组、黄芪甲苷10 mg/ml组和30 mg/ml组、JAK2抑制剂组细胞存活率(P=0.000,0.001,0.049,0.000)和SOD活性(P=0.000,0.002,0.012,0.000)、pJAK2(P=0.003,0.006,0.036,0.002)和pSTAT3(P=0.001,0.002,0.024,0.001)蛋白相对表达量降低,细胞凋亡率(P=0.001,0.001,0.001,0.000)、ROS含量(P=0.000,0.001,0.002,0.000)和LDH活性(P=0.000,0.002,0.038,0.000)升高;与MPP+组相比,黄芪甲苷10 mg/ml组和30 mg/ml组细胞存活率(P=0.016,0.000)和SOD活性(P=0.003,0.001)、pJAK2(P=0.013,0.002)和pSTAT3(P=0.018,0.002)蛋白相对表达量升高,细胞凋亡率(P=0.021,0.008)、ROS含量(P=0.031,0.003)和LDH活性(P=0.001,0.000)降低;与黄芪甲苷10 mg/ml组相比,黄芪甲苷30 mg/ml组细胞存活率(P=0.002)和SOD活性(P=0.027)、pJAK2(P=0.007)和pSTAT3(P=0.006)蛋白相对表达量升高,ROS含量(P=0.019)和LDH活性(P=0.011)降低,JAK2抑制剂组细胞凋亡率(P=0.016)、ROS含量(P=0.030)和LDH活性(P=0.004)升高,SOD活性(P=0.004)、pJAK2(P=0.001)和pSTAT3(P=0.005)蛋白相对表达量降低;与黄芪甲苷30 mg/mlObjective To investigate the effect of astragalosideⅣon the injury of SK-N-SH cell in Parkinson’s disease(PD)induced by 1-methy-4-phenylpyridine(MPP+)and its mechanism.Methods Human neuroblastoma cell line SK-N-SH was cultured to logarithmic growth stage,which were randomized to routine culture(control group),MPP+induction(MPP+group),astragalosideⅣ10 mg/ml+MPP+induction(astragalosideⅣ10 mg/ml group),astragalosideⅣ30 mg/ml+MPP+induction(astragalosideⅣ30 mg/ml group),astragalosideⅣ30 mg/ml+MPP+induction+Janus kinase 2(JAK2)inhibitor AG490(JAK2 inhibitor group).Cell survival was detected by methyl thiazolyl tetrazolium(MTT)assay,cell apoptosis was detected by flow cytometry,reactive oxygen species(ROS)level was detected by DCFH-DA fluorescence probe,lactate dehydrogenase(LDH)and superoxide dismutase(SOD)activities were detected by enzyme method.And the relative expression levels of phosphorylated Janus kinase 2(pJAK2)and phosphorylated signal transducer and activator of transcription 3(pSTAT3)proteins in JAK2-STAT3 signal transducer pathway were detected by Western blotting.Results There were significant differences in cell survival rate(P=0.000),apoptosis rate(P=0.000),ROS content(P=0.000),LDH activity(P=0.000),SOD activity(P=0.003),pJAK2(P=0.000)and pSTAT3(P=0.000)proteins relative expression levels among different SK-N-SH groups.Further pairwise comparison showed that compared with the control group,the cell survival rate(P=0.000,0.001,0.049,0.000),SOD activity(P=0.000,0.002,0.012,0.000),relative expression levels of pJAK2(P=0.003,0.006,0.036,0.002)and pSTAT3(P=0.001,0.002,0.024,0.001)proteins were decreased in MPP+group,astragalosideⅣ10 mg/ml and 30 mg/ml groups,JAK2 inhibitor group,but the cell apoptosis rate(P=0.001,0.001,0.001,0.000),ROS content(P=0.000,0.001,0.002,0.000)and LDH activity(P=0.000,0.002,0.038,0.000)were increased.Compared with MPP+group,the survival rate(P=0.016,0.000),SOD activity(P=0.003,0.001),relative expression of pJAK2(P=0.013,0.002)and pSTAT3(P=0.018,0.002)proteins increa

关 键 词：帕金森病 黄芪 JANUS激酶2 转录激活因子3 肿瘤细胞 培养的 

分 类 号：R285[医药卫生—中药学]