野菊花总黄酮增强细胞自噬促进β淀粉样蛋白清除  

Total flavonoids from Chrysanthemum enhance autophagy and clearance of β-amyloid

作　　者：杨小四 赵欢[2] 曹威[2] 张俊强[2] 陈晓宇[2] Yang Xiaosi;Zhao Huan;Cao Wei;Zhang Junqiang;Chen Xiaoyu(Department of Human Anatomy, Anqing Medical College, Anqing, 246052;Department of Histology and Embryology, Anhui Medical University, Hefei, 230032, China)

机构地区：[1]安庆医药高等专科学校人体解剖学教研室,安庆246052 [2]安徽医科大学组织胚胎学教研室,合肥230032

出　　处：《中国组织化学与细胞化学杂志》2017年第6期537-540,共4页Chinese Journal of Histochemistry and Cytochemistry

基　　金：安徽省高校青年教师重点项目(2006jp100zd);国家自然科学资金面上项目(81373421)

摘　　要：目的探讨野菊花总黄酮(total fl avonoids of Chrysanthemu m,TFC)清除β淀粉样蛋白的作用和分子机制。方法采用CCK-8法观察不同浓度TFC对SH-SY5Y神经细胞株增殖的影响,ELISA法检测TFC对转染过表达淀粉样前体蛋白(APP)和淀粉蛋白前β位分解酶1(BACE1)的CHO细胞培养上清液中Aβ水平的影响,MDC染色检测TFC处理后的神经元内自噬小体的形成,Western blot检测自噬蛋白LC3表达水平。结果 CCK-8法分析显示,TFC对SH-SY5Y神经元活力无明显影响;ELISA检测显示,TFC处理使过表达APP和BACE1的CHO细胞培养上清液中Aβ水平呈浓度依赖性降低;荧光倒置显微镜下观察和Western blot检测发现,TFC处理后SH-SY5Y细胞自噬增加,LC3/LC3自噬标志物蛋白表达增强。结论 TFC可能通过增强细胞自噬而促进Aβ的清除,减少Aβ细胞毒性。Objective To explore the role and molecular mechanism of total flavonoids from Chrysanthemum(TFC)in the clearance ofβ-amyloid(Aβ)peptides.Methods The cytotoxicity of TFC was first evaluated using CCK-8assay in SH-SY5Y cells.Its impact on Aβpeptide secretion was then determined in CHO cells overexpressingβ-amyloid pro-protein(APP)andβ-site APPconverting enzyme1(BACE1),where the Aβlevels in cell culture media was measured by ELISA.In response to TFC treatment in SH-SY5Y cells,autophagy was investigated,including MDC staining of autophagosomes and western blot determination of autophagy marker Light Chain3(LC3).Results CCK-8assay results suggested that TFC was not toxic up to50mg/L in SH-SY5Y cells.In response to TFC treatment,CHO cells showed a dose independent significant reduction on the secreted Aβlevels in the supernatant.The TFC activated autophagy was supported by increased autophagosome counts and LC3Ⅱexpression in TFC treatment group.Conclusion The mechanism that TFC enhances Aβclearance may be through inducing autophagy and decrease the toxicity.

关 键 词：Β淀粉样蛋白 LC3 自噬 野菊花总黄酮 

分 类 号：R332[医药卫生—人体生理学]