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
夏雨薇 1乔云阳 1刘雪薇 1施会敏 1曲高婷 1张爱青 2甘卫华1
作者信息
- 1. 南京医科大学第二附属医院儿科(邮编 210003)
- 2. 南京医科大学第四附属医院儿科
- 折叠
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.
Abstract
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2Key words
diabetic nephropathies/telomeric repeat binding protein 1/tRF-1:30-Gln-CTG-4/renal tubular epithelial cells/inflammatory factors/IKAROS family zinc finger protein 2引用本文复制引用
基金项目
江苏省医学会儿科医学科研专项(SYH-32034-0073)
江苏省医学会儿科医学科研专项(SYH-32034-0085)
南京市卫生科技发展专项(YKK23209)
南京市卫生科技发展专项(YKK23288)
南京市卫生科技发展专项(YKK23289)
出版年
2024
NETL
NSTL
万方数据