生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (3): 16-.doi: 10.3969/j.issn.2095-1736.2023.03.016

• 研究报告 • 上一篇    下一篇

细叶远志皂苷调控PPARγ/PGC-1α信号通路保护D-半乳糖协同Aβ1-42损伤的HT-22细胞

李从婷1,2， 朱国旗1， 陈　彦3， 瞿　艳1， 卞志娟1， 王训翠1   

  1. 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
  • 出版日期:2023-06-18 发布日期:2023-06-19
  • 通讯作者: 王训翠，博士，研究员，硕士生导师，研究方向为神经精神疾病发病机理及中医药防治，E-mail:wangxuncui@163.com
  • 作者简介:李从婷，硕士研究生，研究方向为神经药理学，E-mail:3335117390@qq.com
  • 基金资助:
    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）

Protective effect of tenuifolin on HT-22 cells damaged by D-galactose synergistically with Aβ1-42 via PPARγ/PGC-1α signaling pathway

LI Congting1,2, ZHU Guoqi1, CHEN Yan3, QU Yan1, BIAN Zhijuan1, WANG Xuncui1   

  1. 1. Key Laboratory of Xin’an Medicine, Ministry of Education, Anhui University of Chinese Medicine, Hefei 230038,
    China; 2. College of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China;
    3. Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing 210028, China
  • Online:2023-06-18 Published:2023-06-19

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摘要： 以D-半乳糖协同Aβ1-42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Aβ1-42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、40 μmol/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Aβ1-42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。

关键词: 细叶远志皂苷, Aβ1-42, 线粒体能量代谢, HT-22细胞, PPARγ/PGC-1α通路

Abstract: The AD in vitro model was established by using D-galactose synergistically with Aβ1-42 to induce the damage of HT-22 cells. The purpose of this study was to investigate the role of PPARγ/PGC-1α signaling pathway in D-galactose synergistic Aβ1-42 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, 40 μmol/L) could significantly improve the survival rate of HT-22 cells induced by D-galactose synergistically with Aβ1-42, and reduce the positive rate of β-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-1α in HT-22 cells. The above results suggest that tenuifolin has a protective effect on HT-22 cells damaged by D-galactose synergistically with Aβ1-42 within a certain dose range, and the protective mechanism may be related to up-regulating PPARγ/PGC-1α signaling pathway, enhancing mitochondrial energy metabolism and blocking the inflammatory response.

Key words: tenuifolin, Aβ1-42, mitochondrial energy metabolism, HT-22 cells, PPARγ/PGC-1α signaling pathway

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