摘要
目的探讨蛇床子素上调miRNA-9治疗AD的作用机制。方法基因芯片技术筛选蛇床子素治疗后差异表达的microRNAs;数据库和Cytoscape预测miRNA-9调控的靶基因;脂质体2000建立APP高表达的SH-SY5Y细胞模型,MTT法检测蛇床子素对细胞活力的影响;将miRNA-9抑制剂转入到APP高表达的SH-SY5Y细胞中,RT-PCR法检测各组miR-9、BACE-1 mRNA的表达情况;Western blot方法检测细胞中BACE-1蛋白的表达情况。结果数据库结果显示,蛇床子素调控的miRNA-9可以与BACE-1靶向结合。本实验成功建立APP高表达的SH-SY5Y细胞模型。MTT结果显示,50μmol·L^(-1)蛇床子素对细胞有较好的保护作用。RT-PCR结果显示,蛇床子素可以上调miR-9的表达,抑制剂组miRNA-9表达最低。与模型组相比,蛇床子素可以抑制BACE-1 mRNA和蛋白的表达,且抑制剂组的BACE-1 mRNA和蛋白表达最多。结论蛇床子素治疗阿尔茨海默病可能与上调miRNA-9,从而抑制BACE-1表达有关。
Aim To explore the mechanism of osthole regulating the expression of miR-9 to delay the occurance of Alzheimer’s disease (AD).Methods The miRNA which expressed differently with osthole treatment was selected by microarray;the target gene binding to miRNA-9 was verified by databases and Cytoscape;the SH-SY5Y cells with over expression of APP were established using Lipofectamine2000.The cell viability was determined by MTT assay.The miRNA-9 inhibitor was transfected into cells,and the expression of miRNA-9 and BACE-1 mRNA was determined by RT-PCR;the changes of BACE-1 protein were determined by Western blot.Results The results of database showed that osthole-mediated miRNA-9 was combined with BACE-1 genes.Our lab had established SH-SY5Y cells with over expression of APP.The results of MTT assay showed that 50 μmol·L^-1 osthole had a protective effect on cells.Osthole could increase the expression of miRNA-9,and the expression of miR-9 was lowest in inhibitor group determined by RT-PCR.Osthole decreased the expression of BACE-1 mRNA and protein compared with APP group,and the expression of BACE-1 was highest in inhibitor group.Conclusion Osthole plays a protective role in AD partly through suppressing the expression of BACE-1 by up-regulating miRNA-9.
作者
蔺莹
姚璎珈
梁喜才
时悦
倪颖男
吴雨桐
杨静娴
LIN Ying;YAO Ying-jia;LIANG Xi-cai;SHI Yue;NI Ying-nan;WU Yu-tong;YANG Jing-xian(College of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian Liaoning 116600,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2019年第4期524-529,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81173580)
