艾灸调控SNX5对帕金森病模型小鼠神经元铁死亡的影响  

Moxibustion improves motor function by down-regulating SNX5 and inhibiting ferroptosis in neurons of corpus striatum in mice with Parkinson’s disease

作　　者：高达 黎庆连 王启章 吴春晓 郝小倩 汪晓铃 李萌竹 赵丽君 朱美玲[1,2,3] GAO Da;LI Qing-lian;WANG Qi-zhang;WU Chun-xiao;HAO Xiao-qian;WANG Xiao-ling;LI Meng-zhu;ZHAO Li-jun;ZHU Mei-ling(Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine,Shenzhen 518104,Guangdong Province;Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,Guangdong Province)

机构地区：[1]广州中医药大学,广州510006 [2]深圳市中西医结合医院,广东深圳518104 [3]深圳市中医院,广东深圳518033

出　　处：《针刺研究》2024年第7期736-742,共7页Acupuncture Research

基　　金：广东省自然科学基金项目(No.2022A151510450);深圳市自然科学基金项目(No.JCYJ20220530150212028);深圳市“医疗卫生三名工程”项目(No.SZZYSM202106009);深圳市宝安中医药发展基金会项目(No.2022KJCX-ZJZL-7)。

摘　　要：目的:探讨艾灸调控分选连接蛋白5(SNX5)改善帕金森病(PD)小鼠黑质纹状体铁死亡的作用机制。方法:C57BL/6J小鼠随机分成空白组、假手术组、模型组、艾灸组,每组10只。采用6-羟基多巴胺单侧注入法制备PD模型。艾灸组小鼠予“百会”“四神聪”艾灸,每次20 min,1次/d,每周6次,连续干预4周。干预结束后对各组小鼠进行阿扑吗啡旋转行为检测、爬杆实验,免疫荧光法检测各组小鼠黑质酪氨酸羟化酶(TH)的表达,比色法检测各组小鼠纹状体亚铁离子(Fe2+)、丙二醛(MDA)含量和还原型谷胱甘肽(GSH)/氧化型谷胱甘肽(GSSG)的含量比值,实时荧光定量PCR法和Western blot法检测小鼠纹状体组织SNX5、谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链(FTH1)的mRNA和蛋白表达。结果:与假手术组比较,模型组小鼠每分钟转圈数明显增加(P<0.01),爬杆时间明显延长(P<0.01),黑质TH荧光强度、纹状体GSH/GSSG比值、纹状体GPX4和FTH1的mRNA与蛋白表达降低(P<0.01,P<0.05),纹状体Fe2+与MDA含量、SNX5 mRNA和蛋白表达升高(P<0.01,P<0.05)。与模型组比较,艾灸组小鼠每分钟转圈数明显减少(P<0.01),爬杆时间明显缩短(P<0.01),黑质TH荧光强度、纹状体GSH/GSSG比值、纹状体GPX4和FTH1的mRNA与蛋白表达升高(P<0.01,P<0.05),纹状体Fe2+与MDA含量、SNX5 mRNA和蛋白表达降低(P<0.01,P<0.05)。结论:艾灸可能通过降低SNX5的表达抑制PD模型小鼠神经元铁死亡的发生。Objective To explore the effect of moxibustion on the expression of sorting nexin 5(SNX5),glutathione peroxidase(GPX4)and ferritin heavy chain(FTH1)in the corpus striatum in mice with Parkinson’s disease(PD),so as to explore its mechanisms underlying improvement of PD by ameliorating ferroptosis in the substantia nigra striatum.Methods C57BL/6J mice were randomly divided into normal,sham operation,model,and moxibustion groups,with 10 mice in each group.The PD model was established by unilateral injection of 6-hydroxydopamine(3.5μL)into the right medial forebrain bundle(AP=-1.2 mm,ML=-1.3 mm,DV=-4.75 mm).The mice in the moxibustion group received moxibustion at“Baihui”(GV20)and“Sishencong”(EX-HN1)for 20 min each time,once a day,6 times a week for 4 weeks.After the intervention,mice received apomorphine rotation behavior detection and pole climbing test.The expression of tyrosine hydroxylase(TH)in the substantia nigra was detected by immunofluorescence,the contents of Fe2+,malondialdehyde(MDA),the ratio of glutathione/oxidized glutathione(GSH/GSSG)in the corpus striatum were detected by using photocolorimetric method,and the expression levels of SNX5(endocytosomal protein),GPX4(one of the key targets for inhibiting ferroptosis)and FTH1 proteins and mRNAs in the corpus striatum were detected by Western blot and qPCR,respectively.Results Behavior tests showed that the pole climbing time and number of body rotation were significantly increased in the model group relevant to the sham operation group(P<0.01),and strikingly decreased in the moxibustion group relevant to the model group(P<0.01).The immunofluorescence intensity of TH in the substantia nigra,the ratio of GSH/GSSG,and the expression levels of GPX4 and FTH1 mRNAs and proteins in the corpus striatum were markedly decreased(P<0.01,P<0.05),while the contents of Fe2+and MDA and the expression levels of SNX5 mRNA and protein in the corpus striatum significantly increased in the model group relevant to the sham operation group(P<0.01,P<0.05).Compared wit

关 键 词：艾灸 帕金森病 铁死亡 分选连接蛋白5 

分 类 号：R245[医药卫生—针灸推拿学]