hsa_circ_0006370促进高糖诱导的人肾小管上皮细胞(HK-2)铁死亡

hsa_circ_0006370 promotes ferroptosis in high glucose treated renal tubular epithelial cells(HK-2)

目的探讨hsa_circ_0006370在高糖诱导的肾小管上皮细胞铁死亡中的作用。方法培养人近曲小管上皮细胞HK-2,RT-qPCR检测高糖和爱拉斯汀(Erastin)处理对HK-2细胞中hsa_circ_0006370表达的影响。在HK-2细胞中干扰hsa_circ_0006370表达,将实验分为4组:高糖+si-NC、高糖+si-hsa_circ_0006370、Erastin+si-NC和Erastin+si hsa_circ_0006370,RT-qPCR检测各组细胞中hsa_circ_0006370的表达水平;MTS实验检测各组细胞的增殖水平;相对应试剂盒分别检测各组细胞内铁离子、活性氧(ROS)、乳酸脱氢酶(LDH)、丙二醛(MDA)、胱甘肽过氧化物酶(GSH-Px)水平;Western blot检测各组细胞中的GPX4、xCT、FT、TFR、NRF2、HO-1蛋白表达水平。结果和正常培养组相比,高糖培养组和Erastin培养组细胞中hsa_circ_0006370表达水平显著升高(P均<0.01)。与高糖+si-NC组相比,高糖+si-hsa_circ_0006370组细胞中hsa_circ_0006370相对表达水平显著降低(P<0.01),细胞的增殖能力显著增强(P<0.01),细胞内铁离子、ROS、LDH和MDA活性水平均显著降低,而GSH-Px水平明显增加(P均<0.01),细胞中的GPX4、xCT、FT、NRF2和HO-1蛋白表达水平显著升高,而TFR蛋白表达水平显著降低;与Erastin+si-NC组相比,Erastin+si-hsa_circ_0006370组细胞中hsa_circ_0006370相对表达水平显著降低(P<0.01),细胞的增殖能力显著增强(P<0.01),细胞内铁离子、ROS、LDH和MDA活性水平均显著降低,而GSH-Px水平显著增加(P均<0.01),细胞中的GPX4、xCT、FT、NRF2和HO-1蛋白表达水平显著升高,而TFR蛋白表达水平显著降低。结论hsa_circ_0006370促进高糖诱导的肾小管上皮细胞HK-2铁死亡。

Objective To investigate the effect of hsa_circ_0006370 on promoting ferroptosis in high glucose treated renal tubular epithelial cells HK-2.Methods Human proximal tubular(HK-2)cells were cultured,the effects of high glucose and Erastin treatment on the expression of hsa_circ_0006370 in HK-2 was detected by RT-qPCR.Interfere the expression of hsa_circ_0006370 in HK-2 cells,and the HK-2 cells was divided into four groups:high glucose+si-NC,high glucose+si-hsa_circ_0006370,Erastin+si-NC,Erastin+si-hsa_circ_0006370.The expression level of hsa_circ_0006370 in each group were detected by RT-qPCR.The proliferation of cells in each group was assessed by MTS assay.The levels of ROS,LDH,MDA and GSH-Px in each group were detected with their detection kits,respectively.The protein expression levels of GPX4,xCT,FT,TFR,NRF2,and HO-1 in each group were measured by Western blot.Results Compared with normal culture group,the expression level of hsa_circ_0006370 in high glucose culture group and Erastin culture group were significantly increased(P<0.01).Compared with the high glucose+si-NC group,the relative expression level of hsa_circ_0006370 in high glucose+si-hsa_circ_0006370 group was significantly decreased(P<0.001),the proliferation rate of cells was significantly enhanced(P<0.01),the levels of Fe2+,ROS,LDH,and MDA were significantly decreased,while the GSH-Px level was significantly increased(P<0.01);and the protein expression levels of GPX4,xCT,FT,NRF2,and HO-1 were significantly decreased,while the protein expression level of TFR was significantly increased.Compared with the Erastin+si-NC group,the relative expression level of hsa_circ_0006370 in Erastin+si-hsa_circ_0006370 group was significantly decreased(P<0.01),the proliferation rate of cells was significantly enhanced(P<0.01),the levels of Fe2+,ROS,LDH,and MDA were significantly decreased,while the GSH-Px level was significantly increased(P<0.01);and the protein expression levels of GPX4,xCT,FT,NRF2,and HO-1 were significantly decreased,while the protein expression level of TFR was significantly increased.Conclusion hsa_circ_0006370 promotes ferroptosis in high-glucose treated HK-2 cells.