针刺调控miR-665对帕金森病小鼠BNIP3介导的线粒体自噬的影响  

Effects of Acupuncture Regulation of miR-665 on BNIP3-Mediated Mitochondrial Autophagy in Mice with Parkinson′s Disease

作　　者：班维固[1] 齐辉[1] 卿鹏[2] 滕秀英[1] 陆丽娜 朱莹 许文婷 BAN Weigu;QI Hui;QING Peng;TENG Xiuying;LU Lina;ZHU Ying;XU Wenting(The Second Department of Rehabilitation Medicine,The Second Affiliated Hospital to Heilongjiang University of Chinese Medicine,Harbin Heilongjiang China 150001;Department of Acupuncture and Moxibustion,The First Affiliated Hospital to Jinan University,Guangzhou Guangdong China 510630)

机构地区：[1]黑龙江中医药大学附属第二医院康复医学二科,黑龙江哈尔滨150001 [2]暨南大学附属第一医院针灸科,广东广州510630

出　　处：《中医学报》2024年第6期1138-1143,共6页Acta Chinese Medicine

基　　金：黑龙江中医药管理局面上项目(ZHY2023-104)。

摘　　要：目的:探讨针刺调控miR-665对帕金森病(Parkinson′s disease,PD)小鼠Bcl2/腺病毒E1B相互作用蛋白3(recombinant Bcl2/adenovirus E1B 19kDa interacting protein 3,BNIP3)介导的线粒体自噬的影响。方法:从55只BALB/c小鼠中随机选择10只作为假手术组,其余小鼠皮下注射鱼藤酮溶液(1 mg·kg^(-1)),每日1次,连续14 d。将造模成功的20只小鼠随机分为模型组和针刺组,每组10只。假手术组、模型组小鼠不进行干预,针刺组小鼠给予针刺治疗。采用RT-PCR法检测小鼠纹状体miR-665 mRNA水平,计数小鼠纹状体辅助性T细胞(helper T cells,TH)阳性细胞。通过旷场实验检测小鼠的自主活动情况,采用ELISA法检测小鼠血清白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-8(interleukin-8,IL-8)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平。采用Western blot检测小鼠纹状体mito-LC 3Ⅱ、自噬蛋白5(autophagy protein 5,Atg5)、苄氯素1(recombinant beclin 1,Beclin1)、BNIP3蛋白表达水平。结果:与假手术组比较,模型组、针刺组小鼠纹状体miR-665 mRNA水平升高(P<0.05);与模型组比较,针刺组小鼠纹状体miR-665 mRNA水平降低(P<0.05)。与假手术组比较,模型组、针刺组小鼠脑组织TH阳性细胞数量减少(P<0.05);与模型组比较,针刺组小鼠脑组织TH阳性细胞数量增加(P<0.05)。与假手术组比较,模型组、针刺组小鼠的僵滞时间百分比增加,且运动时间、平均速度、总路程减少(P<0.05);与模型组比较,针刺组小鼠的运动时间、平均速度、总路程增加,且僵滞时间百分比降低(P<0.05)。与假手术组比较,模型组、针刺组小鼠血清IL-1β、IL-6、TNF-α、IL-8水平升高(P<0.05);与模型组比较,针刺组小鼠血清IL-1β、IL-6、TNF-α、IL-8水平降低(P<0.05)。与假手术组比较,模型组、针刺组小鼠纹状体mito-LC 3Ⅱ、Beclin1、Atg5、BNIP3蛋白表达水平升高(P<0.05);与模型组比较,针刺�Objective:To explore the interaction of recombinant Bcl2/adenovirus E1B 19kDa interacting protein 3 in mice with Parkinson′s disease(PD)through acupuncture Effects of BNIP3 on mitochondrial autophagy.Methods:10 out of 55 BALB/c mice were randomly selected as sham operation group,and the other mice were injected subcutaneously with rotenone solution(1 mg·kg^(-1))once a day for 14 days.The 20 mice were randomly divided into model group and acupuncture group,with 10 mice in each group.Mice in sham operation group and model group were not interfered,and mice in acupuncture group were treated with acupuncture.The mRNA level of miR-665 in mouse striatum was detected by RT-PCR,and the helper T cells(TH)positive cells in mouse striatum were counted.The autonomic activity of mice was detected by open field test,and serum interleukin-1β(IL-1β),interleukin-8(IL-8),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.Western blot was used to detect the expression levels of mito-LC 3Ⅱ,autophagy protein 5(Atg5),recombinant beclin 1(Beclin1)and BNIP3 in the striate of mice.Results:Compared with sham operation group,the level of miR-665 mRNA in striatum of mice in model group and acupuncture group was increased(P<0.05).Compared with model group,the level of miR-665 mRNA in striatum of mice in acupuncture group was decreased(P<0.05).Compared with sham operation group,the number of TH positive cells in brain tissue of mice in model group and acupuncture group decreased(P<0.05).Compared with model group,the number of TH positive cells in brain tissue of mice in acupuncture group increased(P<0.05).Compared with sham operation group,the percentage of retarding time increased in model group and acupuncture group,and the movement time,average speed and total distance decreased(P<0.05).Compared with model group,the movement time,average speed,and total distance of mice in acupuncture group increased,and the percentage of stagnant time decreased(P<0.05).Compared with sham operation group,serum level

关 键 词：针刺 帕金森病 miR-665 BNIP3 线粒体自噬 小鼠 

分 类 号：R259.425[医药卫生—中西医结合]