摘要
目的观察微小RNA(miRNA, miR)-135a沉默对急性肾损伤大鼠肾功能保护作用并探讨其分子机制。方法将2018年6月到2019年12月购自河南实验动物中心的60只SD大鼠按照随机数字表法分为对照组、模型组和miR-135a组。miR-135a组通过腹腔注射沉默miR-35a的腺相关病毒1×10^(11) vg/只;对照组和模型组通过腹腔注射对照miRNA腺相关病毒1×10^(11) vg/只。3组大鼠接种15 d后, 模型组和miR-35a组连续15 d注射庆大霉素50 mg/kg建立急性肾损伤模型;对照组连续15 d注射等体积生理盐水。建模成功48 h后处死小鼠, 采用自动生化仪分析大鼠血肌酐和血尿氮水平;采用苏木精-伊红(HE)染色分析肾脏病理学变化;采用荧光定量聚合酶链反应(PCR)分析趋化因子2(CCL2)和趋化因子3(CCL3) mRNA表达水平;采用酶联免疫吸附试验(ELISA)分析3组大鼠外周血炎性因子水平;采用蛋白质印迹法(Western blot)分析miR-135a靶蛋白叉头蛋白转录因子O1(FoxO1)表达水平。计量资料比较采用单因素方差分析。结果 miR-135a组大鼠肾组织病理评分[(5.19±1.38)分]显著低于模型组[(8.09±3.09)分], 差异有统计学意义(t=3.193, P<0.05)。miR-135a组大鼠造模48 h后血肌酐和尿素氮水平[(73.76±9.43)、(12.60±4.01) mmol/L]显著低于模型组[(130.37±10.54)、(29.43±5.12) mmol/L], 差异有统计学意义(t=2.941、3.019, P<0.05)。miR-135a组大鼠肾组织CCL2和CCL3 mRNA表达水平(1.59±0.12、1.73±0.19)显著低于模型组(3.42±0.38、2.51±0.29), 差异有统计学意义(t=2.012、2.240, P<0.05)。miR-135a组大鼠血清白细胞介素(IL)-6和肿瘤坏死因子-α(TNF-α)水平[(165.76±14.36)、(196.250±15.65) pg/ml]显著低于模型组[(252.32±21.44)、(318.02±27.36) pg/ml], 差异有统计学意义(t=3.212、3.937, P<0.05)。miR-135a组大鼠肾组织FoxO1蛋白表达水平(0.29±0.09)显著高于模型组(0.12±0.07), 差异有统计学意义(t=2.719, P<0.05)。结论 miR-135a沉默通过上调FoxO1蛋白表
Objective To observe the protective effect of microRNA(miRNA,miR)-135a silencing on renal function in rats with acute kidney injury and its molecular mechanism.Methods A total of 60 SD rats from June 2018 to December 2019 were randomly divided into control group,model group and miR-135a group.Adeno-associated virus packing miR-135a silencing(1×10^(11) vg)was intraperitoneally injected in miR-135a group.Adeno-associated virus packing miRNA control(1×10^(11) vg)was intraperitoneally injected in control group and model group.After 15 days of inoculation,the model group and miR-35a group were injected with gentamicin(50 mg/kg)for 15 days to establish the acute kidney injury model,and the control group was injected with the same volume of normal saline for 15 days.After 48 h,the mice were killed and the pathological changes of kidney were analyzed by hematoxylin and eosin(HE)staining.The mRNA expression levels of chemokine 2(CCL2)and chemokine 3(CCL3)were analyzed by real-time polymerase chain reaction(PCR).The levels of inflammatory factors in peripheral blood were analyzed by enzyme linked immunosorbent assay(ELISA).The expression levels of forkhead box protein O1(FoxO1)were analyzed by Western blotting.Measurement data were compared by one-way ANOVA.Results Compared with the renal pathological score of model group(8.09±3.09),the pathological score of miR-135a group(5.19±1.38)significantly decreased(t=3.193,P<0.05).Compared with the model group[(130.37±10.54),(29.43±5.12)mmol/L]after modeling for 48 h,the levels of serum creatinine and urea nitrogen in miR-135a group[(73.76±9.43),(12.60±4.01)mmol/L]after modeling for 48 h significantly decreased(t=2.941,3.019,P<0.05).Compared with the mRNA expression levels of CCL2 and CCL3 in model group(3.42±0.38,2.51±0.29),the mRNA expression levels of CCL2 and CCL3 in miR-135a group(1.59±0.12,1.73±0.19)were significantly decreased(t=2.012,2.240,P<0.05).Compared with the model group,the levels of serum interleukin(IL)-6 and tumor necrosis factor-α(TNF-α)in miR-135a
作者
东辛欣
朱桂珍
鲁杨
陈国晓
张士龙
Dong Xinxin;Zhu Guizhen;Lu Yang;Chen Guoxiao;Zhang Shilong(Blood Purification Center,Zhengzhou University People’s Hospital,Henan Provincial People’s Hospital,Zhengzhou 450003,China;Department of Urology,Henan Provincial People’s Hospital,People’s Hospital of Zhengzhou University,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第8期1534-1537,共4页
Chinese Journal of Experimental Surgery
