FGF-1和miR-133a在毛细胞氧化损伤中所致凋亡过程中的表达水平及作用  

The expression and effect of FGF-1 and miR-133a in t-BHP induced cochlear hair cell oxidative damage

作　　者：赵远[1] 冯玉超[1] 苏艺伟 张维森 刘移民[1] ZHAO Yuan; FENG Yu-chao; SU Yi-wei; ZHANG Wei-sen; LIU Yi-ming(Guangzhou Prevention and Treatment Center for Occupational Diseases, Guangzhou , Guangdong 51060, China)

机构地区：[1]广州市职业病防治院,广东广州510620

出　　处：《热带医学杂志》2018年第10期1270-1274,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(81470146);广州市医学重点学科建设项目(穗卫科教【2016】27号);广东省"十二五"医学重点专科(粤卫函【2012】20号);广州市"121人才梯队工程"后备人才项目(穗人社发【2011】167号)

摘　　要：目的观察酸性成纤维细胞生长因子(FGF-1)和miR-133a在HEI-OC1耳蜗毛细胞氧化凋亡过程中的表达,探讨miR-133a对FGF-1的调控机制及其在毛细胞氧化损伤过程中的作用。方法应用叔丁基过氧化氢(t-BHP)致耳蜗毛细胞氧化损伤细胞模型模拟毛细胞受到噪声刺激后的氧化损伤过程,设50、100、200μmol/L三个染毒组和未染毒对照组。提取细胞总RNA和总蛋白,采用qRT-PCR以及Western blot检测miR-133a和FGF-1表达水平,检索生物信息学网站,对FGF-1 3’-UTR区上miR-133a可能作用的靶序列进行预测分析;脂质体转染miR-133amimics入耳蜗毛细胞内,采用q RT-PCR和Western blot分别检测转染后FGF-1 mRNA和蛋白表达水平的变化。结果 qRT-PCR结果显示,t-BHP处理组FGF-1 mRNA的表达为对照组1.32倍(P<0.05)、1.67倍(P<0.05)和2.64倍(P<0.001),且随着染毒浓度的升高呈增加的趋势(F=9.231,P<0.05);同时miR-133a表达均有所降低,分别为对照组的0.81倍(P<0.05)、0.68倍(P<0.05)和0.37倍(P<0.05),且随着染毒浓度的升高呈降低的趋势(F=17.63,P<0.05)。脂质体转染miR-133a mimics,50、100、200μmol/L的转染组miR-133a表达分别为对照组的767、1 499和1 607倍,差异有统计学意义(P<0.05)。miR-133a mimics转染组FGF-1的mRNA和蛋白表达水平均低于对照组,同时miR-133a inhibitors转染组FGF-1的mRNA和蛋白表达水平均高于对照组。软件预测结果显示,在多个物种中靶序列(FGF1-3′UTR上第356-363位核苷酸)在物种之间十分保守,其种子区域完全互补。结论 FGF-1的表达受miR-133a的调控,FGF-1基因的356-363碱基区域可能为miR-133a的识别元件。Objective To investigate the expression of FGF-1 and miR-133 a in cochlear hair cells undergoing oxidativedamage and discuss the regulatory mechanism of miR-133 a on FGF-1 expression and its action in the cochlear hairs cellsdamage process. Methods The t-BHP-induced oxidative damage cell model of cochlear hair cells was used to simulate theoxidative damage process of hair cells after noise stimulation. Set 50,100,200 μmol/L exposure groups and the controlgroup. Total cellular RNA and total protein were extracted. The expression levels of miR-133 a and FGF-1 were detected byqRT-PCR and Western-blot. The possible miR-133 a target sequences on the 3′-UTR region of FGF-1 were predicted bybioinformatic tools. miR-133 a mimics were transfected into the cochlear hair cells by liposomes. The expressions of FGF-1 mRNA and protein were detected by q RT-PCR and Western-blot,respectively. Results The results of qRT-PCR showedthat the expression of FGF-1 in t-BHP treatment group was 1.32 times（P〈0.05）,1.67 times（P〈0.05）and 2.64 times（P〈0.001）of those in the control group,and the level was increased with the increase in the concentration of the drug（F=9.231,P〈0.05）. At the same time,the expression of miR-133 a was decreased,which was 0.81 times（P〈0.05）,0.68 times（P〈0.05）and 0.37 times of the control group（P〈0.05）,and there was a tendency to decrease with increasingconcentration of the drug（F=17.63,P〈0.05）. The FGF-1 levels were increased for 767 times,1 499 times and 1 607 timesin the cells transfected with miR-133 a mimics at 50,100 and 200 μmol/L,respectively（P〈0.05）. The m RNA and proteinexpression levels of FGF-1 in miR-133 a mimics transfection group were lower than those in the control group,and themRNA and protein expression levels of FGF-1 in miR-133 a inhibitors transfected group were higher than those in the control group. The result of software forecast shows that the terget sequences（356-363 nucleotide on 3′ UTR of FGF-1） areconservative and compatible

关 键 词：毛细胞 氧化损伤 FGF-1 miR-133a 

分 类 号：R446.6[医药卫生—诊断学]