安神定志方对阿尔茨海默病细胞模型miR-103a-3p及Tau蛋白磷酸化的影响  

Effects of Anshen Dingzhi Prescription on miR-103a-3p and Tau Protein Phosphorylation in Alzheimer Disease Cell Model

作　　者：王欣波[1] 齐明明[2] 邵音 赵宇[1] 聂双莲[1] 朴勇洙[1] WANG Xinbo;QI Mingming;SHAO Yin;ZHAO Yu;NIE Shuanglian;PIAO Yongzhu(The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040 [2]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《中国中医药信息杂志》2023年第6期106-111,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：黑龙江省自然科学基金面上项目(H2018063);黑龙江省中医药科研项目(21102190005)。

摘　　要：目的观察安神定志方对阿尔茨海默病细胞模型miR-103a-3p及其介导的Tau蛋白磷酸化的影响,探讨其可能作用机制。方法采用β淀粉样蛋白_(25-35)诱导PC12细胞构建阿尔茨海默病体外模型,将细胞分为空白组、模型组、安神定志方组、过表达组和安神定志方+过表达组(联合组),采用安神定志方含药血清和miR-103a-3p模拟剂进行干预。流式细胞仪检测细胞凋亡,试剂盒检测细胞上清液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)含量,RT-qPCR检测细胞miR-103a-3p mRNA表达,Western blot检测细胞脑源性神经营养因子(BDNF)、酪氨酸激酶受体B(TrkB)、p-TrkB、Tau、p-Tau蛋白表达。结果与空白组比较,模型组PC12细胞各时点细胞活力明显降低,凋亡率明显增加,LDH活性和MDA含量明显增加,SOD和GSH-Px活性明显降低,miR-103a-3p mRNA表达明显升高,p-Tau蛋白表达明显升高,BDNF、p-TrkB蛋白表达明显降低(P<0.01);与模型组比较,安神定志方组和联合组PC12细胞各时点细胞活力明显升高,凋亡率明显降低,LDH活性和MDA含量明显降低,SOD和GSH-Px活性明显增加,miR-103a-3p mRNA表达明显降低,p-Tau蛋白表达明显降低,BDNF、p-TrkB蛋白表达明显升高(P<0.05,P<0.01)。结论安神定志方能通过下调miR-103a-3p表达激活BDNF/TrkB信号通路,从而抑制Tau蛋白磷酸化,发挥对PC12细胞的保护作用。Objective To observe the effects of Anshen Dingzhi Prescription on tau protein phosphorylation mediated by miR-103a-3p in Alzheimer disease cell model;To discuss its mechanism of action.Methods In vitro model of Alzheimer disease established by PC12 cells induced by Aβ_(25-35).The cells were divided into blank group,model group,Anshen Dingzhi Prescription group,overexpression group,and Anshen Dingzhi Prescription+overexpression group(combination group),and were intervened with Anshen Dingzhi Prescription containing serum and miR-103a-3p simulated agent.Apoptosis was detected by flow cytometry;the kit was used to detect the activities of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)and thecontent of malondialdehyde(MDA)in cell supernatant;RT-qPCR was used to detect cellular miR-103a-3p mRNA expression;Western blot was used to detect the expressions of brain-derived neurotrophic factor(BDNF),tyrosine kinase receptor B(TrkB),p-TrkB,Tau and p-Tau protein.Results Compared with the blank group,the cell viability of PC12 cells in the model group significantly decreased at each time point,the apoptosis rate significantly increased,the LDH activity and MDA content significantly increased,the activities of SOD and GSH-Px significantly decreased,the mRNA expression of miR-103a-3p significantly increased,the protein expression of p-Tau significantly increased,and the protein expressions of BDNF and p-TrkB significantly decreased(P<0.01).Compared with the model group,the cell viability of PC12 cells in Anshen Dingzhi Prescription group and the combination group significantly increased at each time point,the apoptosis rate significantly decreased,the LDH activity and MDA content significantly decreased,the activities of SOD and GSH-Px significantly increased,the mRNA expression of miR-103a-3p significantly decreased,the protein expression of p-Tau significantly decreased,and the protein expressions of BDNF and p-TrkB significantly increased(P<0.05,P<0.01).Conclusion Anshen Dingzhi Pres

关 键 词：安神定志方 阿尔茨海默病 PC12细胞 miR-103a-3p TAU蛋白磷酸化 

分 类 号：R285.5[医药卫生—中药学]