miR-137对Parkin诱导的线粒体自噬的影响  

Effect of miR-137 on Parkin-induced mitophagy

作　　者：李文[1] 胡喆[2] 林智君[3] 杨玥[2] 陈煜森[3] 钟望涛[3] 林春霞[3] 赵斌[1] 冯杜[1] 

机构地区：[1]广东医学院附属医院神经病学研究所 广东省衰老相关心脑血管疾病重点实验室,湛江524001 [2]广东医学院附属医院麻醉科,湛江524001 [3]广东医学院附属医院神经内科,湛江524001

出　　处：《新医学》2015年第5期283-288,共6页Journal of New Medicine

基　　金：国家自然科学青年基金项目(31301104;31401182);广东省教育厅育苗工程项目(2013LYM_0035);广东省卫生厅项目(A2014484);广东医学院青年基金项目(XQ1328;XQ1336)

摘　　要：目的探讨miR-137对Parkin诱导的线粒体自噬的影响。方法将miR-137 mimics和miR-137 inhibitor分别转染进He La细胞8 h后,再转染p EGEPC1-Parkin,24 h之后加羰基-氰-对-三氟甲氧基本腙(FCCP)处理4 h或者不加FCCP,利用免疫荧光和蛋白免疫印迹技术分析miR-137对线粒体自噬的影响。用实时定量PCR方法检测帕金森病患者与健康对照者血液样本中miRNAs的表达情况。结果在FCCP作用下,Parkin的表达促进细胞自噬分子LC3由LC3Ⅰ型变为LC3Ⅱ型,Parkin蛋白从胞浆转位至线粒体。而细胞先经外源过表达miR-137处理之后,Parkin引起的细胞自噬受到显著抑制(P=0.003)。miR-137在帕金森病患者血液中的表达水平显著高于健康对照组(P<0.001)。结论在FCCP作用下,miR-137抑制Parkin介导的线粒体自噬。miR-137在帕金森病患者血液样本中高表达,提示miR-137可能通过调控Parkin介导的线粒体自噬,参与帕金森病的发生。Objective To investigate the effect of miR-137 upon Parkin-induced mitophagy.Me-thods miR-137 mimics and inhibitors were transfected into Hela cells for 8 h.pEGFPC1-Parkin was then transfected for another 24 h.Hela cells were treated with or without carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone （FCCP）for 4 h before being harvested.Immunofluorescence and Western blot were used for detec-ting the effect of miR-137 on mitophagy.Expression of miRNAs in blood sample from patients with Parkinson＇s disease （PD）and healthy donors were measured with Real-time quantitative PCR.Results Under the inter-vention of FCCP,the expressed Parkin promoted the conversion of LC3Ⅰ into LC3Ⅱ and translocated the Par-kin protein from cytoplasm to mitochondria.After in vitro cell treatment with over-expressed miR-137,Parkin-induced mitophagy was significantly inhibited （P =0.003）.The expression of miR-137 in the blood of PD pa-tients was significantly higher than that of normal controls （P 〈0.001）.Conclusions miR-137 could sup-press Parkin-mediated mitophagy in the presence of FCCP.The high expression level of miR-137 in PD patients indicates that miR-137 may be involved in the incidence of PD via regulating Parkin-mediated mitophagy.

关 键 词：MIRNA 线粒体自噬 PARKIN 帕金森病 羰基-氰-对-三氟甲氧基本腙 

分 类 号：R742.5[医药卫生—神经病学与精神病学]