【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。更多还原

【Abstract】 Objective To investigate the effect of miR-137 upon Parkin-induced mitophagy. Methods miR-137 mimics and inhibitors were transfected into Hela cells for 8 h. p EGFPC1-Parkin was then transfected for another 24 h. Hela cells were treated with or without carbonyl cyanide p-trifluoromethoxyphenylhydrazone( FCCP) for 4 h before being harvested. Immunofluorescence and Western blot were used for detecting 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 intervention of FCCP,the expressed Parkin promoted the conversion of LC3Ⅰ into LC3Ⅱ and translocated the Parkin protein from cytoplasm to mitochondria. After in vitro cell treatment with over-expressed miR-137,Parkininduced mitophagy was significantly inhibited( P = 0. 003). The expression of miR-137 in the blood of PD patients was significantly higher than that of normal controls( P < 0. 001). Conclusions miR-137 could suppress 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.更多还原