小檗碱通过NF-κB相关通路抑制AGEs诱导的小鼠小胶质细胞活化的实验研究  

Experimental Study on the Effects of Berberine on Mouse Microglia Cultured with Advanced Glycation end Products(AGEs)via the NF-κB Signaling Pathway

作　　者：万瑾舒 顾蓓[2] 刘玉琴[2] 南凤尾 张孟仁[1] Wan JinshuGu Bei;Liu Yuqin(Department of Traditional Chinese Medicine,Peking Union Medical College Hospital,Peking Union Medical College and Chinese Academy of Medical Sciences,Bejing 100730,China)

机构地区：[1]中国医学科学院/北京协和医学院北京协和医院中医科,100730 [2]中国医学科学院基础医学研究所协和细胞资源中心,北京100730

出　　处：《医学研究杂志》2020年第3期105-108,共4页Journal of Medical Research

摘　　要：目的 探讨小檗碱对AGEs诱导的小鼠小胶质细胞活化的干预作用和机制.方法 将细胞分为CON组、AGEs组、小檗碱和BSA阴性对照组,MTT法检测细胞活力,倒置相差显微镜下观察细胞的形态变化,ELISA法检测细胞上清液中TNF-α的分泌情况,Western blot法检测细胞内CD68、RAGE、pNF-κB p65蛋白的表达.结果 AGEs在不改变细胞活力的情况下,对小胶质细胞的形态有改变作用,并且可以显著上调细胞上清液中的炎性细胞因子TNF-α的分泌量(P=0.000),增加CD68、RAGE及pNF-κB p65蛋白的表达(P<0.05).小檗碱可以改善AGEs诱导的小胶质细胞形态改变,显著下调小鼠小胶质细胞CD68和pNF-κB p65蛋白的表达(P<0.05),对TNF-α的分泌量没有影响.结论 小檗碱可以抑制AGEs诱导的小鼠小胶质细胞的活化,其作用可能与NF-κB介导的炎性信号通路有关.Objective To explore the effects of AGEs on the activation of mouse microglia and the effects of berberine on mouse microglia cultured with AGEs. Methods The microglia were divided into four groups: control group, AGEs group, berberine group and BSA group. The morphological changes of microglia were observed by inverted phrase contrast microscope and cell viability was evaluated by MTT assays. Meanwhile, we adopted Enzyme-linked immunosorbent assay(ELISA) to determine the concentration of TNF-α in the supernatant of cultured microglia and Western blot analysis to semiquantitatively detect the expression of CD68,RAGE and pNF-κB p65. Results AGEs(300μg/ml) could remarkably up-regulate the expression of CD68,RAGE and pNF-κB p65(P<0.05),thus increase the secretion of TNF-α in microglia(P=0.000),while had no effect on the viability of microglia. Meanwhile, berberine could down-regulate the expression of CD68 and pNF-κB p65(P<0.05) in mouse microglia cultured with AGEs(300μg/ml) for 24 h,while had no effect on the expression of RAGE and the concentration of TNF-α in the cell supernatant. Conclusion Berberine can inhibit the activation of microglia cultured with AGEs and the mechanism may be involved with the NF-κB signaling pathway.

关 键 词：小檗碱 AGES 小胶质细胞 神经炎性反应 糖尿病认知功能障碍 

分 类 号：R587.1[医药卫生—内分泌]