当归含药血清通过调控miR-129表达对阿尔茨海默病细胞模型的保护作用  

Protective effect of Angelica medicated serum on Alzheimer’s disease cell model by regulating the expression of miR-129

作　　者：李云莹 刘敬 姚新宇[2] Li Yunying;Liu Jing;Yao Xinyu(Affiliated Hospital of Heze Medical College,Heze Shandong274000;不详)

机构地区：[1]山东省菏泽医学专科学校附属医院,274000 [2]河北省邢台市人民医院

出　　处：《卒中与神经疾病》2021年第4期389-395,409,共8页Stroke and Nervous Diseases

基　　金：河北省重点研发计划项目(项目编号为182777151)。

摘　　要：目的探讨当归含药血清通过调控微小RNA-129(MicroRNA-129,miR-129)表达对阿尔茨海默病PC12细胞的保护作用。方法分别以1.5 g/kg当归药液及等量生理盐水对SD(Sprague Dawley)大鼠灌胃处理,配制成10%含药血清及正常血清培养液;将PC12细胞分为空白组、模型组、药物组、药物+Control小干扰RNA(Small interference RNA,siRNA)组、药物+miR-129 siRNA组,除空白组外,其余各组均用30μL的Aβ25-35溶液诱导细胞损伤24 h构建阿尔茨海默病(Alzheimer's disease,AD)细胞模型,空白组及模型组用空白血清培养液培养,其余各组用10%含药血清培养液培养,培养48 h后收集各组细胞观察细胞形态学;实时荧光定量聚合酶链反应(Quantitative real-time polymerase chain reaction,qRT-PCR)法检测其中miR-129表达水平,采用甲基噻唑蓝(Methyl thiazolyl tetrazolium,MTT)法检测各组细胞的增殖抑制率;流式细胞仪检测各组细胞的凋亡及周期情况;蛋白免疫印迹(Western blot,WB)检测细胞凋亡相关因子蛋白的表达水平;酶联免疫吸附(Enzyme-linked immunoSorbent assay,ELISA)试剂盒检测乳酸脱氢酶(Lactate dehydrogenase,LDH)、丙二醛(Malondialdehyde,MDA)、超氧化物歧化酶(T-superoxide dismutase,T-SOD)水平。结果与空白组比较,模型组PC12细胞贴壁功能受损,miR-129、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白水平、S期及G2/M期细胞数目、T-SOD水平降低(P<0.05),细胞抑制率、凋亡率、半胱氨酸蛋白酶-3(Cysteinyl aspartate specific proteinase,Caspase-3)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax),G0/G1期细胞数目、LDH,MDA水平升高(P<0.05);与模型组比较,药物组细胞贴壁功能改善,miR-129,Bcl-2蛋白水平、S期及G2/M期细胞数目、T-SOD水平升高(P<0.05),细胞抑制率、凋亡率、Caspase-3,Bax,G0/G1期细胞数目、LDH,MDA水平下降(P<0.05);与药物组、药物+Control siRNA组比较,细胞贴壁功能受损,miR-129,Bcl-2蛋白水平、S期及G2/M期细胞数目、T-SOD�Objective To investigate the mechanisms underlying the protective effect of Angelicamedicated serum onPC12 cells.Methods Sprague dawleyrats were treated with 1.5g/kg Angelica solution or the same amount of normal saline;PC12 cells were divided into blank group,model group,drug group,drug+Control small interference RNA(siRNA)group and drug+miR-129 siRNA group.Except for the blank group,30μL Aβ25-35solution was added for 24 hto establish AD cell model in other groups.Cells in the blank group and model group were cultured in blank serum medium,others were cultured in 10%medicated serum medium.After 48 hours of culture,the cells were collected for morphological observation.The expression of MiR-129 was detected by quantitative real-time polymerase chain reaction(qRT-PCR)method.Cell proliferation inhibition rate was detected by methyl thiazolyl tetrazolium(MTT)assay.Cell apoptosis and cell cycle were detected by flow cytometry.The expression of apoptotic related factors was detected by Western blotting(WB).Lactate dehydrogenase(LDH),malondialdehyde(MDA)and T-SuperoxideDismutase(T-SOD)level were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with those in the blank group,the adhesion function of PC12 cells in the model group was impaired,the levels of miR-129 and B-cell lymphoma-2(Bcl-2)protein,numbers of S phase and G2/M phase cells,T-SOD were lower(P<0.05),and the cell inhibition rate,apoptosis rate,cysteinyl aspartate specific proteinase(Caspase-3),Bcl-2 associated X protein(Bax),number of G0/G1 phase cells,LDH,MDA were higher(P<0.05).Compared with those in the model group,cell adhesion function in the drug group was improved,the levels of miR-129 and Bcl-2 protein,numbers of S phase and G2/M phase cells,T-SOD were higher(P<0.05).While the cell inhibition rate,apoptosis rate,caspase-3,Bax,number of G0/G1 phase cells,LDH,MDA were lower(P<0.05).Compared with those in drug group and drug+Control siRNA group,cell adhesion function was impaired,the levels of miR-129 and Bcl-2 protein,numbers of S

关 键 词：当归 含药血清 微小RNA-129 阿尔茨海默病 

分 类 号：R742[医药卫生—神经病学与精神病学]