新型长链非编码RNA XLOC_032768在顺铂诱导急性肾损伤小鼠中的作用

The role of a novel long non-coding ribonucleicacid XLOC_032768 in cisplatin induced acute renal injury in mice

目的探讨新型长链非编码RNA(lncRNA)XLOC_032768在顺铂诱导急性肾损伤小鼠中的作用及其机制。方法30 mg/kg顺铂腹腔注射小鼠(鼠龄范围为6~8周,体重20~25 g,购自江苏卡文斯实验动物中心)建立急性肾损伤模型,通过小鼠肾脏全转录组测序和反转录-聚合酶链反应(RT-PCR)筛选出新型lncRNA进行实验研究。将24只雄性C57小鼠随机分为4组:对照+lncRNA对照组、顺铂+lncRNA对照组、对照+lncRNA治疗组、顺铂+lncRNA治疗组。血清检测肌酐(Cr)和尿素氮(BUN)含量;苏木精-伊红(HE)染色观察肾脏组织病理学变化评估肾小管损伤;RT-PCR检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6表达。组间两样本均数的比较采用t检验及单因素方差分析。结果全转录组测序结果显示新型lncRNA-XLOC_032768在顺铂诱导的急性肾损伤小鼠肾脏中表达显著降低,RT-PCR结果亦显示顺铂组lncRNA XLOC_032768表达量较对照组显著降低(0.99±0.01比0.41±0.05,t=11.850,P<0.05),差异有统计学意义。与对照+lncRNA对照组比较,顺铂+lncRNA对照组肌酐(26.67±2.12比93.83±3.50,t=20.670,P<0.05)、尿素氮显著升高(20.83±1.70比52.17±2.36,t=10.780,P<0.05),加重肾小管损伤(0.17±0.17比2.50±0.22,t=8.370,P<0.05)及TNF-α表达增高(0.95±0.22比3.35±0.15,t=16.060,P<0.05)、IL-1β(0.98±0.01比2.68±1.23,t=13.340,P<0.05)、IL-6(0.98±0.01比3.53±0.17,t=15.300,P<0.05),差异有统计学意义。与顺铂+lncRNA对照组比较,顺铂+lncRNA-XLOC_032768治疗组显著降低肌酐(65.83±2.39,t=6.610,P<0.05)及尿素氮表达(33.00±1.83,t=6.420,P<0.05),改善肾小管损伤(1.33±0.21,t=3.790,P<0.05)及降低TNF-α(1.83±0.17,t=6.330,P<0.05)、IL-1β(2.03±0.13,t=3.560,P<0.05)、IL-6表达(2.55±0.18,t=4.050,P<0.05),差异有统计学意义。结论lncRNA-XLOC_032768通过减少炎性反应保护顺铂诱导的急性肾损伤。

Objective To explore the effect of a novel long non-coding RNA(lncRNA)XLOC_032768 on cisplatin induced acute renal injury in mice.Methods The model of acute renal injury was established by intraperitoneal injection of 30 mg/kg cisplatin.A new gene was selected by performing differentially expressed genes(DEGs)of the transcriptome data and real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)experiment.Twenty-four male C57 mice were randomly divided into 4 groups:control group,cisplatin group,cisplatin+lncRNA control group,cisplatin+lncRNA treatment group.The levels of serum creatinine(Cr)and ureanitrogen(BUN)were measured.Renal tubular injury was estimated by hematoxylin and eosin(HE)staining.The level of tumor necrosis factor alpha(TNF)-α,interleukin(IL)-1βand IL-6 was detected by PCR.The SPSS 17.0 software was used for statistical analysis.Results We found the expression of lncRNA XLOC_032768 was significantly repressed by cisplatin treatment by performing DEGs of the transcriptome data,which was also validated by RT-qPCR experiment(0.99±0.01 vs.0.41±0.05,t=11.850,P<0.05).Compared with the control group,the serum Cr(26.67±2.12 vs.93.83±3.50,t=20.670,P<0.05),and BUN(20.83±1.70 vs.52.17±2.36,t=10.780,P<0.05)levels,the renal tubular injury score(0.17±0.17 vs.2.50±0.22,t=8.370,P<0.05),the level of TNF-α(0.95±0.22 vs.3.35±0.15,t=16.060,P<0.05),IL-1β(0.98±0.01 vs.2.68±1.23,t=13.340,P<0.05),IL-6(0.98±0.01 vs.3.53±0.17,t=15.300,P<0.05)in cisplatin group was increased.Compared with the cisplatin group,the serum Cr(65.83±2.39,t=6.610,P<0.05)and BUN(33.00±1.83,t=6.420,P<0.05)levels,the renal tubular injury score(1.33±0.21,t=3.790,P<0.05),the level of TNF-α(1.83±0.17,t=6.330,P<0.05),IL-1β(2.03±0.13,t=3.560,P<0.05),IL-6(2.55±0.18,t=4.050,P<0.05)in cisplatin+lncRNA group was decreased.Conclusion A novel lncRNA XLOC_032768 protects cisplatin-induced acute renal injury by reducing inflammatory response.