MiR-29a靶向调控瞬时受体电位通道家族4在睾丸缺血再灌注损伤中对GC-1细胞增殖及凋亡的影响及作用机制  

Effect and mechanism of miR-29a target regulating TRPV4 on proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury

作　　者：宁金卓[1] 程帆[1] 余伟民[1] 饶婷[1] 阮远[1] 袁润[1] Ning Jinzhuo;Cheng Fan;Yu Weimin;Rao Ting;Ruan Yuan;Yuan Run(Department of Urology,the People's Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,430060

出　　处：《国际泌尿系统杂志》2021年第1期124-128,共5页International Journal of Urology and Nephrology

基　　金：武汉大学自主科研项目(2042019kf0059)。

摘　　要：目的研究miR-29a通过靶向调控瞬时受体电位通道家族4(TRPV4)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞增殖和凋亡的影响及作用机制。方法建立睾丸GC-1细胞缺氧复氧模型,RT-PCR检测不同复氧损伤时间点miR-29a和TRPV4的表达水平;分别采用MTT实验、流式细胞术检测转染miR-29a对GC-1细胞增殖和凋亡能力的变化;荧光素酶实验证实miR-29a和TRPV4的靶向关系,Western blot法测定转染miR-29a后细胞中TRPV4的蛋白表达水平。结果miR-29a随着复氧损伤时间的增加,其表达显著降低,而TRPV4的表达明显增加;过表达miR-29a能显著促进GC-1细胞的增殖能力,抑制细胞凋亡水平。沉默miR-29a可使GC-1细胞的增殖能力明显减弱,凋亡水平增加;荧光素酶报告实验表明TRPV4是miR-29a的下游靶基因。过表达miR-29a能明显抑制GC-1细胞的TRPV4表达,而沉默miR-29a能显著促进TRPV4表达。结论MiR-29a可以通过靶向调控TRPV4来改变GC-1细胞的增殖和凋亡能力,从而影响睾丸IRI的发生发展。Objective To investigate the effect and mechanism of miR-29a on the proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury(IRI)by targeting TRPV4.Methods The hypoxia reoxygenation model of testicular GC-1 cells was established,RT-PCR was used to detect the expression of miR-29a and TRPV4 at different reoxygenation injury time points.The effect of transfected miR-29a on the proliferation and apoptosis of GC-1 cells was measured by MTT method and flow cytometry.Luciferase reporter was used to verify the relationship between miR-29a and TRPV4,and Western blot method was used to examine the protein expression level of THPV4 after miR-29a was transfected.Results The expression of miH-29a was significantly decreased with the increasing reoxygenation injury time,and the TRPV4 expression was increased notably.The overexpression of miR-29a promoted the proliferation of GC-1 cells and inhibit the level of apoptosis.The silence of miR-29a decreased the proliferation of GC-1 cells and increased the level of apoptosis.The luciferase reporter showed TRPV4 was the downstream target gene of miR-29a.The overexpression of miR-29a significantly inhibited the TRPV4 expression,while the silence of miR-29a promoted the TRPV4 expression of GC-1 cells.Conclusions MiR-29a could change the proliferation and apoptosis of GC-1 cells by targeting TRPV4,thus affecting the development of testicular IRI.

关 键 词：睾丸 再灌注损伤 瞬时受体电位通道 GC-1细胞 Mir-29a 

分 类 号：R726.9[医药卫生—儿科]