摘要
目的:探讨滋肾活血方通过调节线粒体自噬促进双侧颈总动脉永久性结扎(2-VO)模型大鼠海马CA1区神经元新生的作用机制。方法:2-VO法建立血管性痴呆大鼠模型,随机将60只SD大鼠分为假手术组、2-VO模型组、盐酸多奈哌齐组、滋肾活血方低、中、高剂量组(8.9、17.8、35.6 g·kg^(-1))。Morris水迷宫实验检测各组逃避潜伏期、跨越平台次数;透射电镜观察海马CA1区线粒体自噬情况;实时荧光定量聚合酶链式反应(Real-time PCR)检测海马CA1区PTEN诱导激酶1(Pink1)、帕金蛋白(Parkin)mRNA的表达;蛋白免疫印迹法(Western blot)检测海马CA1区线粒体自噬信号通路相关蛋白Parkin、抗增殖蛋白2(PHB2)、线粒体融合蛋白2(Mfn2)、动力学相关蛋白1(Drp1)蛋白的表达;Brdu法检测CA1区神经元再生情况。结果:与假手术组比较,2-VO模型组大鼠学习、空间记忆能力明显下降(P<0.05),海马CA1区见线粒体结构受损,自噬溶酶体形成,Parkin、Pink1 mRNA及Parkin、PHB2、Drp1蛋白明显上调(P<0.05),Mfn2蛋白表达量明显降低(P<0.05),海马CA1区神经元新生明显减少(P<0.05)。与模型组比较,滋肾活血方干预后可明显提高2-VO模型大鼠的学习、空间记忆能力(P<0.05),增加海马CA1区自噬小体数量并改善线粒体结构,进一步上调海马CA1区Parkin、Pink1 mRNA及Parkin、PHB2、Drp1蛋白表达(P<0.05,P<0.01),下调Mfn2蛋白表达(P<0.05,P<0.01),增加海马CA1区神经元新生数量(P<0.05,P<0.01)。结论:滋肾活血方对血管性痴呆大鼠海马CA1区神经元新生的促进作用与Pink1/Parkin信号通路介导的线粒体自噬有关。
Objective:To investigate the effect of Zishen Huoxue prescription on promoting neurogenesis in hippocampal CA1 region of vascular dementia(VD)rats by regulating mitophagy.Method:The 2-VO method was used to establish the VD rat model and 60 SD rats were randomly divided into sham operation group,model group,donepezil hydrochloride group,and Zishen Huoxue prescription low-dose(8.9 g·kg^(-1)),medium-dose(17.8 g·kg^(-1))and high-dose(35.6 g·kg^(-1))groups.Morris water maze test was performed to detect the escape latency and the number of crossing platform in each group.The expression of phosphatase and tensin homology-induced kinase 1(PINK1)and Parkinson protein(Parkin)mRNA in hippocampal CA1 region was detected by Real-time fluorescent quantitative polymerase chain reaction(Realtime PCR).Western blot was used to determine the expression of mitochondrial autophagy signaling pathwayrelated proteins Parkin,prohibitin 2(PHB2),mitofusin 2(Mfn2)and dynamin-related protein 1(Drp1)in hippocampal CA1 region.The neurogenesis in hippocampal CA1 region was tested by Brdu method.Result:Compared with the conditions in the sham operation group,the learning and spatial memory abilities of the model group were decreased(P<0.05),with damaged mitochondrial structure and autolysosome formation in the hippocampal CA1 region.The expressions of Parkin,Pink1 mRNA and Parkin,PHB2,and Drp1 proteins were up-regulated(P<0.05),while the expression of Mfn2 protein and the neuronal regeneration in hippocampal CA1 region were reduced(P<0.05,P<0.05).Compared with the conditions in the model group,Zishen Huoxue prescription enhanced the learning and spatial memory abilities of VD rats(P<0.05),increased the number of autophagosomes in hippocampal CA1 region and improved the mitochondrial structure.The expression of Parkin,Pink1 mRNA and Parkin,PHB2,and Drp1 proteins in hippocampal CA1 region was up-regulated(P<0.05,P<0.01)while the expression of Mfn2 protein was down-regulated(P<0.05,P<0.01).The number of new neurons in hippocampal CA1 region was
作者
赵梓婷
伍大华
张秀丽
谢乐
刘桐赫
曾珊珊
施佳仪
姚婷
谢瑶
李映辰
ZHAO Ziting;WU Dahua;ZHANG Xiuli;XIE Le;LIU Tonghe;ZENG Shanshan;SHI Jiayi;YAO Ting;XIE Yao;LI Yinchen(Hunan University of Chinese Medicine,Changsha 410208,China;Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha410006,China;Science and Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第20期45-52,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81874462)
湖南省重点研发计划项目(2020SK2123)
湖南省教育厅优秀青年基金项目(19B426)。
关键词
血管性痴呆
CA1区
神经元新生
线粒体分裂蛋白
认知障碍
滋肾活血方
vascular dementia
CA1 region
neurogenesis
mitochondrial fission protein
cognitive impairment
Zishen Huoxue prescription