微小RNA通过SIRT1调控小鼠耳蜗老化HEI-OC1细胞凋亡的研究  

miR-96 regulates apoptosis of mouse cochlear aging HEI-OC1 cells via SIRT1

作　　者：袁佳添 谢奇伟 王玥 程鹏[2] 王耀文[2] YUAN Jiatian;XIE Qiwei;WANG Yue;CHENG Peng;WANG Yaowen(Department of Otolaryngogy,Beilun People's Hospital,Ningbo,Zhejiang,315826,China;Department of Otolaryngogy Head and Neck Surgery,Ningbo First Hospital,Ningbo,Zhejiang,315000,China;Medical College of Ningbo University,Ningbo,Zhejiang,315000,China)

机构地区：[1]宁波市北仑区人民医院耳鼻咽喉科,浙江宁波315826 [2]宁波市第一医院耳鼻咽喉头颈外科,浙江宁波315000 [3]宁波大学医学院,浙江宁波315000

出　　处：《中国耳鼻咽喉头颈外科》2023年第2期84-88,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：浙江省医药卫生科技项目(2019KY584);浙江省中医药科技计划项目(2023ZL648)。

摘　　要：目的构建小鼠耳蜗毛细胞株HEI-OC1(house ear institute-organ of Corti 1)老化模型,探索微小RNA-96(miR-96)可否通过SIRT1调控老化HEI-OC1凋亡。方法CCK-8筛选D-半乳糖浓度。流式细胞仪检测细胞凋亡,Western blot检测p53、p38、caspase-3蛋白表达。q RT-PCR检测miR-96、SIRT1 mRNA表达,Western blot检测SIRT1蛋白表达。miR-96模拟物转染HEI-OC1细胞,qRT-PCR检测miR-96、SIRT1 mRNA表达,Western blot检测SIRT1、caspase-3蛋白表达。结果HEI-OC1细胞在15 mg/ml D-半乳糖条件下培养72 h后,凋亡较对照组显著增加(P<0.01),且p53、p38(P<0.05)及caspase-3(P<0.0001)蛋白表达明显增加。老化组miR-96与SIRT1 mRNA和蛋白表达均明显减低(P<0.05)。miR-96过表达后,SIRT1 mRNA及蛋白较对照组表达升高,caspase-3蛋白表达降低(P<0.05)。结论老化HEI-OC1细胞中miR-96、SIRT1 mRNA与蛋白的表达均下降。miR-96可通过上调SIRT1抑制HEI-OC1细胞的凋亡。miR-96、SIRT1可能成为保护老年性聋耳蜗毛细胞的作用靶点。OBJECTIVE To explore whether miR-96 could regulate the apoptosis of aging HEI-OC1(house ear institute-organ of Corti 1)through SIRT1.METHODS D-galactose concentration was screened by CCK8.Apoptosis was detected by flow cytometry,and protein expressions of p53,p38 and caspase-3 were detected by Western blot.The mRNA expressions of miR-96 and SIRT1 were detected by qRT-PCR,and the protein expressions of SIRT1 were detected by Western blot.The miR-96 mimics were transfected into HEI-OC1 cells,mRNA expressions of miR-96 and SIRT1 were detected by qRT-PCR,and protein expressions of SIRT1 and caspase-3 were detected by Western blot.RESULTS HEI-OC1 cells were cultured in 15 mg/ml D-galactose for 72 hours.The apoptosis of HEI-OC1 cells was significantly increased compared with the control group(P<0.01),and the protein expressions of p53,p38(P<0.05)and caspase-3(P<0.0001)were significantly increased.The expression of miR-96、SIRT1 mRNA and protein in HEI-OC1 cells of aging group were significantly decreased(P<0.05).After miR-96 overexpression,SIRT1 mRNA and protein expression were increased compared with that of the control group,while caspase-3 protein expression was decreased(P<0.05).CONCLUSION The expression of miR-96 as well as SIRT1 mRNA and protein in aging HEI-OC1 cells were decreased.miR-96 can inhibit the apoptosis of HEI-OC1 cells by upregulating SIRT1.miR-96 and SIRT1 may become targets for protecting cochlear hair cells in presbycusis.

关 键 词：老年性聋 细胞凋亡 miR-96 SIRT1 HEI-OC1 

分 类 号：R764.436[医药卫生—耳鼻咽喉科]