黄芩素通过miR-141-3p/sirt7介导的线粒体自噬发挥帕金森病神经保护作用的机制研究  

Mechanism of baicalin's neuroprotective effect on Parkinson's disease through miR-141-3p/sirt7 mediated mitochondrial autophagy

作　　者：李耀杨 金旭东 高玉峰 Li Yaoyang;Jin Xudong;Gao Yufeng(Department of Neurology,the Huhehot of Inner Mongolia Hospital,BeijingUniversity Tumor Hospital,Huhehot O10010,China)

机构地区：[1]内蒙古医科大学附属人民医院神经内科,呼和浩特010010

出　　处：《脑与神经疾病杂志》2024年第5期290-296,共7页Journal of Brain and Nervous Diseases

基　　金：内蒙古自治区自然科学基金项目[2017MS(LH)0821]。

摘　　要：目的探讨黄芩素(Baicalein)对帕金森病(PD)神经保护的可能机制。方法在本研究中,采用6-羟基多巴胺(6-OHDA)诱导小鼠的PD模型,研究黄岑素的保护作用机制。首先,对黄芩素干预后的PD模型小鼠进行神经学评分,高效液相色谱电化学法检测纹状体多巴胺含量,TUNEL和Fluoro-Jade B染色检测凋亡细胞及神经元。采用自噬抑制剂3-甲基腺嘌呤(3-MA)抑制PD模型小鼠线粒体自噬,通过Jc-1染色检测黄芩素干预后的PD模型小鼠的线粒体膜电位,通过Western blot检测自噬相关蛋白LC3、P62。在PD模型体内转染agomiR-141-3p,通过qRT-PCR检测转染效率,通过Jc-1和Wb检测线粒体膜电位和自噬相关蛋白的变化情况。通过Starbase网站(https://starbase.sysu.edu.cn/)、双荧光素酶切报告实验在293T细胞中分析miR-141-3p和SIRT7的靶向关系,qRT-PCR检测SIRT7在PD模型体内的表达水平。结果PD严重影响了小鼠的神经功能,下调了线粒体膜电位水平及线粒体自噬相关蛋白LC3的表达水平、上调了P62的表达水平。而黄芩素提高了PD模型小鼠的神经学评分,一定程度上还原了多巴胺与神经元的数目,恢复了线粒体膜电位水平及LC3和P62的表达水平;黄芩素抑制了PD小鼠miR-141-3p的表达,上调miR-141-3p可以逆转黄芩素促线粒体膜电位上升、LC3 z-Ⅱ/LC3-Ⅰ比值上升及P62表达下降;而miR-141-3p可以靶向负调控SIRT7,通过下调SIRT7同样可以逆转黄芩素促线粒体膜电位上升、LC3-Ⅱ/LC3-Ⅰ比值上调及P62表达下调;黄芩素可以使SIRT7表达上调,而过表达miR-141-3p可以抑制SIRT7的表达。结论黄芩素通过下调miR-141-3p靶向负调控SIRT7介导线粒体自噬,从而对PD神经发生保护。Objective This article aims to explore the possible mechanism of baicalin in the neuroprotection of Parkinson's disease(PD).Methods In this study,we used a PD model induced by 6-hydroxydopamine(6-OHDA)in mice to investigate the protective mechanism of Baicalein.First of all,we performed neurology scoring on PD model mice after baicalein intervention,detected dopamine content in striatum by high performance liquid chromatography electrochemical method,and detected apoptotic cells and neurons by TUNEL and Fluoro Jade B staining.Autophagy of PD model mice was inhibited by autophagy inhibitor 3-methyladenine(3-MA).Mitochondrial membrane potential of PD model mice after baicalein intervention was detected by Jc-1 staining,and autophagy related proteins LC3 and P62 were detected by Western blot.AgomiR-141-3p was transfected into PD model,and the transfection efficiency was detected by qRT-PCR.The changes of mitochondrial membrane potential and autophagy related proteins were detected by Jc-1 and WB.Through the Starbase website(https:/starbase.sysu.edu.cn/),the targeting relationship between miR-141-3p and SIRT7 was analyzed in 293T cells by double Luciferase digestion report experiment,and the expression level of SIRT7 in PD model was detected by qRT-PCR.Results PD surgery seriously affected the neural function of mice,down regulated the level of mitochondrial membrane potential and the expression level of mitochondrial autophagy related protein LC3,and up regulated the expression level of P62.Baicalein increased the neurology score of PD model mice,reduced the number of dopamine and neurons to a certain extent,and restored the level of mitochondrial membrane potential and the expression levels of LC3 and P62.Baicalein inhibited the expression of miR-141-3p in PD mice,and upregulation of miR-141-3p could reverse the increase of mitochondrial membrane potential,the increase of LC3-II/LC3-I ratio and the decrease of P62 expression induced by baicalein.However,miR-141-3p can negatively regulate SIRT7 by targeting.By downr

关 键 词：帕金森病 黄芩素 微小核糖核酸-141-3p 沉默调节蛋白7 线粒体自噬 6-羟基多巴胺 靶向调控 

分 类 号：R742.5[医药卫生—神经病学与精神病学]