过表达miR-29a对人椎间盘退变髓核细胞凋亡的影响  

Overexpression of MicroRNA-29a regulates nucleus pulposus cells apoptosis in human intervertebral disc degeneration

作　　者：刘晓潭[1] 田林强[1] 王宏伟[1] 郭志豪[1] 

机构地区：[1]新乡医学院第三附属医院骨二科,河南新乡453003

出　　处：《重庆医学》2016年第14期1893-1895,共3页Chongqing medicine

摘　　要：目的探讨miR-29a在人椎间盘退变髓核(NP)细胞凋亡的调控作用。方法体外培养椎间盘退变NP细胞,构建重组真核表达质粒cherry/miR-29a,脂质体Lipofectamine转染进入NP细胞,应用反转录实时定量PCR(RT-qPCR)检测NP细胞中miR-29a的表达变化,Western blot法检测凋亡相关蛋白Caspase-3前体的表达变化,流式细胞仪检测NP细胞凋亡水平变化。结果 NP细胞转染重组质粒cherry/miR-29a48h后,NP细胞中miR-29a的表达水平较NP细胞和转染空质粒的NP细胞中的miR-29a明显升高(P<0.05);凋亡相关蛋白Caspase-3前体的水平则明显下降(P<0.05);NP细胞凋亡水平明显升高(P<0.05)。结论在椎间盘NP细胞中过表达miR-29a能促进NP细胞凋亡,其机制可能是通过Caspase-3途径起作用。Objective To investigate the role of MicroRNA-29a（miR-29a）on nucleus pulposus cells apoptosis in human intervertebral disc degeneration.Methods Intervertebral disc degeneration nucleus pulposus cells was isolated,cells were transfected with recombinant plasmid cherry/miR-29 aby lipofectamine method,and then RT-qPCR was used to measure the expressive level of miR-29;the protein expressive level of Caspase-3was detected by Western blot and flow cytometry（FCM）was applied to detect the cells apoptosis.Results NP cells transfer to the recombinant plasmid cherry/miR-29 a.The miR-29 aexpression level of NP cell was significantly higher compared with the blank and empty plasmid group（P〈0.05）;the apoptosis-related protein Caspase-3precursor level significantly decreased（P〈0.05）while the NP cell apoptosis level significantly increased（P〈0.05）after 48 hours.Conclusion The overexpression of miR-29 amight regulate nucleus pulposus cells apoptosis of human intervertebral disc degeneration,and the mechanisms among it might be concerned with Caspase-3pathway.

关 键 词：椎间盘 细胞凋亡 Caspase-3 过表达 miR-29a 椎间盘退变NP细胞 

分 类 号：R681.53[医药卫生—骨科学]