【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。更多还原

【Abstract】 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.更多还原