tRF-1:30对高糖诱导的肾小管上皮细胞炎性因子表达的影响  

Effect of tRF-1:30 on the expression of inflammatory factors in renal tubular epithelial cells induced by high glucose

作　　者：夏雨薇 乔云阳 刘雪薇 施会敏 曲高婷 张爱青[2] 甘卫华[1] XIA Yuwei;QIAO Yunyang;LIU Xuewei;SHI Huimin;QU Gaoting;ZHANG Aiqing;GAN Weihua(Department of Pediatric Nephrology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210003,China;Department of Pediatric Nephrology,the Fourth Affiliated Hospital of Nanjing Medical University)

机构地区：[1]南京医科大学第二附属医院儿科,210003 [2]南京医科大学第四附属医院儿科

出　　处：《天津医药》2024年第6期561-566,共6页Tianjin Medical Journal

基　　金：江苏省医学会儿科医学科研专项基金项目(SYH-32034-0073,SYH-32034-0085);南京市卫生科技发展专项资金项目(YKK23209,YKK23288,YKK23289)。

摘　　要：目的探讨tRF-1:30(tRF-1:30-Gln-CTG-4)对高糖(HG)诱导的肾小管上皮细胞(RTECs)中炎性因子表达的影响及分子机制。方法将小鼠RTECs分为Control组、HG组、HG+tRF-1:30 mimic组、HG+tRF-1:30 NC组、HG+si-IKZF2组(IKAROS家族锌指2,tRF-1:30抑制剂)、HG+si-NC组。实时荧光定量PCR检测tRF-1:30、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、单核细胞趋化蛋白-1(MCP-1)和IKZF2 mRNA的水平。酶联免疫吸附试验检测炎性因子水平,Western blot检测IKZF2蛋白表达水平,双萤光素酶报告实验验证tRF-1:30和IKZF2的关系。结果在HG诱导的RTECs中炎性因子的表达水平显著升高,而tRF-1:30表达水平显著降低。过表达tRF-1:30显著降低HG诱导的RTECs中炎性因子的表达水平。IKZF2在HG诱导的RTECs中显著高表达,进一步敲低IKZF2可抑制炎性因子的释放,而过表达tRF-1:30后IKZF2的表达水平下调。双萤光素酶报告实验进一步验证tRF-1:30与IKZF2可能存在靶向关系。结论过表达tRF-1:30可能通过负向调控IKZF2的表达进而抑制HG诱导的RTECs炎性因子的释放。Objective To investigate the effect and molecular mechanism of tRF-1:30-Gln-CTG-4(tRF-1:30)on the expression of inflammatory factors in high glucose(HG)-induced renal tubular epithelial cells(RTECs).Methods RTECs were divided into the control group,the HG group,the HG+tRF-1:30 mimic group,the HG+tRF-1:30 negative control(NC)group,the HG+si-IKZF2 group and the HG+si-NC group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of tRF-1:30,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1)and IKAROS family zinc finger protein 2(IKZF2).Enzyme-linked immunosorbent assay(ELISA)was used to detect levels of TNF-α,IL-6 and MCP-1.Protein expression of IKZF2 was detected by Western blot assay.Dual-luciferase reporter assay was used to detect the targeting relationship between tRF-1:30 and IKZF2.Results The expression levels of inflammatory factors were elevated in HG-induced RTECs,and the expression level of tRF-1:30 was decreased(P<0.05).Overexpression of tRF-1:30 significantly decreased expression levels of inflammatory factors in HG-induced RTECs(P<0.05),and the expression level of IKZF2 was significantly increased(P<0.05).Further knockdown of IKZF2 can inhibit the release of inflammatory factors,and the expression level of IKZF2 was down-regulated after overexpression of tRF-1:30.Double luciferase reporting experiment further verified the possible targeting relationship between tRF-1:30 and IKZF2.Conclusion Overexpression of tRF-1:30 inhibits the expression of inflammatory factors in HG-induced RTECs by target binding and negatively regulating the expression of IKZF2.

关 键 词：糖尿病肾病 端粒重复序列结合蛋白质1 tRF-1:30-Gln-CTG-4 肾小管上皮细胞 炎性因子 IKAROS家族锌指2 

分 类 号：R587.2[医药卫生—内分泌]