基于线粒体质量控制系统观察六味地黄丸对老龄小鼠的神经保护作用  

Neuroprotective effect of liuwei dihuangwan on aged mice based on the mitochondrial quality control system

作　　者：王世界 杨晶[1] 马贺 王璐瑶 张紫娟[2] 苏运芳 张振强[1] WANG Shijie;YANG Jing;MA He;WANG Luyao;ZHANG Zijuan;SU Yunfang;ZHANG Zhenqiang(Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046,China;School of Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046)

机构地区：[1]河南中医药大学中医药科学院,郑州450046 [2]河南中医药大学医学院,郑州450046

出　　处：《中国比较医学杂志》2023年第6期24-34,共11页Chinese Journal of Comparative Medicine

基　　金：河南省高校科技创新团队支持计划资助(21IRTSTHN026);河南省科技攻关项目(212102310314,222102310280)。

摘　　要：目的观察六味地黄丸对老龄小鼠海马区线粒体质量控制系统的影响及神经保护机制。方法将50只20月龄C57BL/6J老龄小鼠按照每组10只进行随机分组,包括模型组,六味地黄丸低、中、高剂量组,多奈哌齐组,另选10只3月龄C57BL/6J野生小鼠为正常组。各组给药60 d后,进行行为学实验,免疫荧光检测胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、肿瘤坏死因子(tumor necrosis factor,TNF-α)的表达,透射电镜观察线粒体的健康程度及自噬小体的数量,Western blot检测线粒体质量控制系统相关蛋白。结果高剂量组与模型组比较,行为学实验结果显示六味地黄丸可以提高小鼠的学习记忆能力,增强运动能力,减轻焦虑;免疫荧光实验显示GFAP、TNF-α表达下降,电镜观察发现正常线粒体数量增多;Western blot实验结果显示PTEN诱导激酶1(PTEN induced putative kinase 1,PINK1)、帕金蛋白(PARKIN)、自噬相关3号染色体基因(autophagy related 3,ATG3)、自噬相关7号染色体基因(autophagy related 7,ATG7)、微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3β,LC3B)、线粒体融合蛋白2(mitochondrial fusion protein 2,MFN2)、线粒体内源发动蛋白1(optic atrophy 1,OPA1)、线粒体转录因子A(recombinant transcription factor A,mitochondrial,TFAM)、γ共激活因子-1α(peroxisome proliferator-activated receptorγcoactivator 1α,PGC-1α)蛋白的表达上升,泛肽结合蛋白1(sequestosome protein 1,P62)、线粒体动力相关蛋白1(dynamin-related protein 1,DRP1)、TNF-α蛋白的表达下降。结论六味地黄丸可以提高小鼠的认知学习能力,减轻小鼠海马区的神经炎症,改善线粒体质量控制系统。Objective To observe the effect of liuwei dihuangwan on the mitochondrial quality control system in the hippocampus of aged mice and its neuroprotective mechanism.Methods Fifty 20-month-old C57BL/6J mice were randomly divided into five groups with 10 mice in each group,including a model group liuwei dihnangwan low,medium,and high dose groups,donepezil group.Ten 3-month-old C57BL/6J mice were selected as the normal group.After 60 days of treatment,behavioral tests were carried out.Immunofluorescence was used to assess TNF-αand GFAP expression.Transmission electron microscopy was used to observe mitochondria and the number of autophagosomes.Proteins of the mitochondrial quality control system were assessed by Western blot.Results Compared with the model group,behavioral tests showed that liuwei dihuangwan improved learning and memory abilities,enhanced the exercise ability,and reduced anxiety.Immunofluorescence showed decreases in TNF-αand GFAP expression.Transmission electron microscopy showed that the number of normal mitochondria was increased.PINK1,PARKIN,ATG3,ATG7,LC3B,MFN2,OPA1,TFAM,and PGC-1αprotein expression was upregulated in the high-dose group,while P62,DRP1,and TNF-αprotein expression was downregulated.Conclusions Liuwei dihuangwan improves the cognitive learning ability of mice,reduces neuroinflammation in the hippocampus,and improves the quality control system of mitochondria.

关 键 词：六味地黄丸 脑老化 线粒体质量控制系统 神经炎症 

分 类 号：R-33[医药卫生]