tRF-1:32-Glu-TTC-1介导高糖诱导肾小管上皮细胞焦亡的机制研究  

Mechanism of tRF-1:32-Glu-TTC-1 in mediating high glucose-induced pyroptosis in renal tubular epithelial cells

作　　者：刘雪薇 黄婵 夏雨薇 宁蓓蓓 施会敏 曲高婷 张爱青[2] 甘卫华[1] LIU Xuewei;HUANG Chan;XIA Yuwei;NING Beibei;SHI Huimin;QU Gaoting;ZHANG Aiqing;GAN Weihua(Department of Pediatric Nephrology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210003,China;Department of Pediatric Nephrology,the Fourth Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210031)

机构地区：[1]南京医科大学第二附属医院小儿肾内科,江苏南京210003 [2]南京医科大学第四附属医院小儿肾内科,江苏南京210031

出　　处：《徐州医科大学学报》2024年第12期859-864,共6页Journal of Xuzhou Medical University

基　　金：国家自然科学基金青年基金(82400864);江苏省自然科学基金面上项目(BK20211385);江苏省医学会儿科医学科研专项基金[SYH-32034-0085(20230032),SYH-32034-0073(20230020)];南京市卫生科技发展专项资金项目(YKK23209、YKK23288、YKK23289);南医大二附院卓越人才培养计划(789ZYRC202090251)。

摘　　要：目的探究tRF-1:32-Glu-TTC-1在高糖(HG)诱导的肾小管上皮细胞(RTECs)焦亡中的作用及相关机制。方法用高糖(35 mmol/L)干预RTECs,RT-PCR法检测tRF-1:32-Glu-TTC-1的表达,扫描电镜观察细胞形态变化,RT-PCR法检测NLRP3、GSDMD在mRNA水平上的表达变化,Western blot法检测NLRP3、caspase-1、GSDMD-N在蛋白水平上的表达变化;采用tRF-1:32-Glu-TTC-1抑制剂转染RTECs并用RT-PCR法检测转染效率,RT-PCR及Western blot法检测焦亡相关指标在mRNA及蛋白水平表达变化,Western blot法检测通路蛋白p-PI3K/PI3K、p-Akt/Akt、p-FoxO3a/FoxO3a的表达水平。结果与Control组相比,HG组tRF-1:32-Glu-TTC-1表达增加,扫描电镜下可见细胞焦亡发生,焦亡相关指标转录上调及蛋白水平表达增加;抑制表达tRF-1:32-Glu-TTC-1组焦亡相关指标在mRNA及蛋白水平低于HG组及抑制剂空载组(HG+NC inhibitor组);与Control组相比,HG组p-PI3K/PI3K、p-Akt/Akt、p-FoxO3a/FoxO3a蛋白减少;抑制表达tRF-1:32-Glu-TTC-1组p-PI3K/PI3K、p-Akt/Akt、p-FoxO3a/FoxO3a蛋白水平高于HG组及抑制剂空载组。结论tRF-1:32-Glu-TTC-1可能通过负向调控PI3K/Akt/FoxO3a信号通路促进高糖诱导的RTECs焦亡。Objective To investigate the mechanism of tRF-1:32-Glu-TTC-1 in mediating high glucose(HG)-induced pyroptosis in renal tubular epithelial cells(RTECs).Methods RTECs were treated with high glucose(35 mmol/L),and the expression of tRF-1:32-Glu-TTC-1 was detected by RT-PCR.The morphological changes of these cells were observed by scanning electron microscopy(SEM).The mRNA levels of NLRP3 and GSDMD were detected by RT-PCR,and the protein levels of NLRP3,caspase-1,and GSDMD-N were measured by Western blot.RTECs were transfected with tRF-1:32-Glu-TTC-1 inhibitor,and the transfection efficiency was detected by RT-PCR.The mRNA and protein levels of pyroptosis-related indicators were detected by Western blot.The protein levels of p-PI3K/PI3K,p-Akt/Akt,and p-FoxO3a/FoxO3a were measured by Western blot.Results Compared with the Control group,the HG group showed increased expression of tRF-1:32-Glu-TTC-1,and pyroptosis was observed by SEM.The mRNA and protein levels of pyroptosis-related indicators was upregulated.The tRF-1:32-Glu-TTC-1 inhibition group showed lower mRNA and protein levels of pyroptosis-related indicators than the HG group and the inhibitor blank vector group.Compared with the Control group,decreased levels of p-PI3K/PI3K,p-Akt/Akt,and p-FoxO3a/FoxO3a proteins were observed in the HG group.The tRF-1:32-Glu-TTC-1 inhibition group presented higher protein levels of p-PI3K/PI3K,p-Akt/Akt,and p-FoxO3a/FoxO3a than the HG group and the inhibitor blank vector group.Conclusions tRF-1:32-Glu-TTC-1 may promote HG-induced pyroptosis in RTECs through negatively regulating the PI3K/Akt/FoxO3a signaling pathway.

关 键 词：tRF-1:32-Glu-TTC-1 糖尿病肾病 肾小管上皮细胞 焦亡 信号通路 

分 类 号：R692.6[医药卫生—泌尿科学]