细叶远志皂苷调控PPARγ/PGC-1α信号通路保护D-半乳糖协同Aβ_(1-42)损伤的HT-22细胞  

Protective effect of tenuifolin on HT-22 cells damaged by D-galactose synergistically with Aβ_(1-42) via PPARγ/PGC-1αsignaling pathway

作　　者：李从婷 朱国旗[1] 陈彦[3] 瞿艳 卞志娟 王训翠[1] LI Congting;ZHU Guoqi;CHEN Yan;QU Yan;BIAN Zhijuan;WANG Xuncui(Key Laboratory of Xin’an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Hefei 230038,China;College of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing 210028,China)

机构地区：[1]安徽中医药大学新安医学教育部重点实验室,合肥230038 [2]安徽中医药大学药学院,合肥230012 [3]江苏省中医药研究院,南京210028

出　　处：《生物学杂志》2023年第3期16-21,共6页Journal of Biology

基　　金：安徽省教育厅高校自然科学研究项目(KJ2021A0584);安徽省科技厅自然科学基金项目(2108085MH316);安徽中医药大学人才支持计划项目(2021rcyb008)。

摘　　要：以D-半乳糖协同Aβ_(1-42)诱导HT-22细胞损伤建立AD体外模型,旨在研究PPARγ/PGC-1α信号通路在D-半乳糖协同Aβ_(1-42)诱导HT-22细胞损伤中的作用及细叶远志皂苷的干预机制。采用MTT法检测细胞存活率;台盼蓝染料排斥实验检测细胞死亡率;β-半乳糖苷酶染色检测细胞衰老情况;Mito-Tracker荧光标记观察线粒体损伤程度;罗丹明123染色流式细胞仪检测线粒体膜电位变化;比色法检测超微量总ATP酶含量改变;Western Blot法检测PPARγ、PGC-1α、COX-2和NF-κB蛋白的表达;ELISA法检测炎性因子TNF-α和IL-6的含量。结果显示,与模型组相比,细叶远志皂苷(10、20、40μmol/L)能显著提高D-半乳糖协同Aβ1-42诱导的HT-22细胞存活率,降低β-半乳糖苷酶阳性表达,增加细胞内ATP酶活性,阻止线粒体膜电位下降,抑制炎症因子TNF-α和IL-6水平的增加,下调NF-κB和COX-2蛋白的表达同时上调HT-22细胞中PPARγ、PGC-1α蛋白的表达。结果提示细叶远志皂苷在一定剂量范围内对D-半乳糖协同Aβ_(1-42)损伤的HT-22细胞具有保护作用,其保护机制可能与上调PPARγ/PGC-1α信号通路、增强线粒体能量代谢、阻止炎症反应有关。The AD in vitro model was established by using D-galactose synergistically with Aβ_(1-42) to induce the damage of HT-22 cells.The purpose of this study was to investigate the role of PPARγ/PGC-1αsignaling pathway in D-galactose synergistic Aβ_(1-42) induced HT-22 cell injury and the intervention mechanism of tenuifolin.MTT assay was used to detect cell viability;Trypan blue dye exclusion assay was used to detect cell death rate;β-galactosidase staining was used to detect cell aging;Mito-Tracker fluorescent labeling to observe mitochondrial damage;Rhodamine 123 staining was applied to detect changes in mitochondrial membrane potential by flow cytometry;Colorimetry was used to detect changes in ultra-trace total ATPase content;Western Blotting was used to detect PPARγ,PGC-1α,COX-2 and NF-κB protein expression.The content of inflammatory factors such as TNF-α,IL-6 were detected by ELISA method.The results showed that compared with the model group,tenuifolin(10,20,40μmol/L)could significantly improve the survival rate of HT-22 cells induced by D-galactose synergistically with Aβ_(1-42),and reduce the positive rate ofβ-galactosidase,increase intracellular ATPase activity,prevent the decrease of mitochondrial membrane potential,inhibit the increase of inflammatory factors such as TNF-α,IL-6 levels,down-regulate the expression of NF-κB and COX-2 and up-regulate the expression of PPARγ,PGC-1αin HT-22 cells.The above results suggest that tenuifolin has a protective effect on HT-22 cells damaged by D-galactose synergistically with Aβ_(1-42) within a certain dose range,and the protective mechanism may be related to up-regulating PPARγ/PGC-1αsignaling pathway,enhancing mitochondrial energy metabolism and blocking the inflammatory response.

关 键 词：细叶远志皂苷 Aβ_(1-42) 线粒体能量代谢 HT-22细胞 PPARγ/PGC-1α通路 

分 类 号：R285[医药卫生—中药学]