miR-21-3p/p53信号通路对缺血/再灌注损伤小鼠的肾脏保护作用及其机制  

Protective effects and the mechanism of miR-21-3p/p53 signaling pathway on renal function in mice with renal ischemia/reperfusion injury

作　　者：雷小林 刘燕秀 张婵 LEI Xiaolin;LIU Yanxiu;ZHANG Chan(Department of Nephrology,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710000,China;不详)

机构地区：[1]西安交通大学第一附属医院肾脏内科,西安710000 [2]吉安市中心人民医院消化内科,江西吉安343000 [3]西北工业大学医学研究院,西安710072

出　　处：《实用医学杂志》2023年第19期2475-2482,共8页The Journal of Practical Medicine

基　　金：国家自然科学基金-青年科学基金项目(编号:82200386);中国博士后科学基金-面上项目(编号:2022M712590);陕西省自然科学基础研究计划项目-青年项目(编号:2022JQ-881)。

摘　　要：目的观察miR-21-3p对肾缺血/再灌注(ischemia/reperfusion,I/R)损伤小鼠的影响,并探讨其肾脏保护作用是否与通过miR-21-3p/p53信号通路而调控凋亡水平有关。方法体内实验随机分为Sham、I/R、I/R+agomir、I/R+agomir+PIF、I/R+PIF共5组。建立小鼠肾缺血/再灌注(I/R)损伤模型;构建过表达miR-21-3p(agomiR-21-3p)小鼠,同时给予p53抑制剂Pifithrin-α(PIF)处理;采用双荧光素酶报告基因实验验证miR-21-3p与p53的互作;检测小鼠血清中肾功能指标小鼠胱抑素C(Cys C)、尿素氮(BUN)、肌酐(Scr)的含量,以及炎症指标肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、超敏C反应蛋白(hs-CRP)含量水平;此外,TUNEL染色观察肾细胞凋亡情况,并检测肾组织(细胞)匀浆中细胞凋亡相关基因p53、Bax、Bcl2、Caspase-3的表达水平。结果与Sham组比较,I/R组小鼠肾细胞miR-21-3p表达明显降低;而与I/R组相比,I/R+agomir组、I/R+PIF组和I/R+agomir+PIF组miR-21-3p表达均明显升高(P<0.05),而p53表达趋势相反;双荧光素酶报告基因实验证实miR-21-3p与p53存在互作关系;与Sham组比较,其他肾组织细胞凋亡率均升高;但是与I/R组相比,I/R+agomir组、I/R+PIF组和I/R+agomir+PIF组凋亡率降低,I/R+agomir+PIF组损伤最低(P<0.05)。结论miR-21-3p可以通过靶向下调p53表达来抑制凋亡的发生,从而减轻肾I/R小鼠的肾损伤,发挥肾脏保护作用。Objective To observe the effect of miR-21-3p on mice with renal ischemia/reperfusion(I/R)injury and to explore whether the renal protective effect is related to the regulation of apoptosis levels through miR-21-3p/p53 signaling pathway.Methods In vivo experiments were randomly divided into five groups:Sham,I/R,I/R+agomir,I/R+agomir+PIF,and I/R+PIF.Renal I/R injury model in vivo was established,overexpressing of miR-21-3p(agomiR-21-3p)in mice were constructed and treated with the p53 inhibitor Pifithrin-α(PIF).Dualluciferase reporter assay was used to verify the interaction between miR-21-3p and p53.Serum levels of cystain C(Cys C),blood urea nitrogen(BUN)and serum creatinine(Scr)were measured to reflect renal function of mice.Serum oxidative stress and inflammation indices were also determined,including tumor necrosis factor-alpha(TNF-α),Interleukin-1β(IL-1β),and high-sensitive c-reactive protein(hs-CRP).In addition,apoptosis rate of renal cells was visualized by TUNEL staining and the expression of apoptosis b-cell lymphoma-2(Bcl-2),Bcl-2 assaciated X protein(Bax),p53 and caspase-3 were measured too.Results Compared with Sham group,the expression of miR-21-3p in I/R mice was significantly decreased.Compared with I/R group,the expression of miR-21-3p in the I/R+agomir group,the I/R+PIF group and the I/R+agomir+PIF group was significantly increased(P<0.05),while the trend of p53 expression was opposite.Dual-luciferase reporter assay confirmed the interaction between miR-21-3p and p53.Compared with Sham group,the rate of cell apoptosis in all other renal tissues was increased;however,compared with the I/R group,the I/R+agomir,the I/R+PIF group and I/R+agomir+PIF groups were decreased,and the I/R+agomir+PIF group was the lowest(P<0.05).Conclusions miR-21-3p could inhibit the development of apoptosis by targeting p53 expression,thus alleviating renal injury in mice with renal I/R.

关 键 词：miR-21-3p 缺血/再灌注损伤 p53 细胞凋亡 

分 类 号：R363.14[医药卫生—病理学]