摘要
目的探讨miR-488-3p调控环磷酸鸟苷-腺苷酸合成酶(cGAS)-干扰素基因刺激因子(STING)通路对肾小管上皮细胞损伤的影响。方法将人肾小管上皮细胞HK-2分为对照组、缺氧复氧(HR)组、抑制对照组、miR-488-3p抑制组、cGAS-STING通路激活剂(RocA)组、miR-488-3p抑制+RocA组。用实时定量PCR检测HK-2细胞中miR-488-3p的表达水平;用CCK-8法检测细胞活力;用ELISA检测HK-2细胞上清液中白细胞介素(IL)-10、IL-1β、肿瘤坏死因子α(TNF-α)水平;用DCFH-DA法检测细胞中活性氧(ROS)平均荧光强度。结果与对照组比较,HR组HK-2细胞中miR-488-3p表达水平、上清液中IL-1β和TNF-α水平、细胞中ROS平均荧光强度、丙二醛(MDA)水平、细胞凋亡率及cleaved caspase-3、Bax、cGAS、STING蛋白表达水平升高,细胞活力、上清液中IL-10水平、细胞中超氧化物歧化酶(SOD)水平降低(P<0.05);与HR组、抑制对照组比较,miR-488-3p抑制组HK-2细胞中miR-488-3p表达、上清液中IL-1β和TNF-α水平、细胞中ROS平均荧光强度、MDA水平、细胞凋亡率及cleaved caspase-3、Bax、cGAS、STING蛋白表达水平降低,细胞活力、上清液中IL-10水平、细胞中SOD水平升高(P<0.05)。结论下调miR-488-3p表达可抑制HR诱导的HK-2细胞炎症、氧化应激及细胞凋亡,减轻细胞损伤的机制可能与抑制cGAS-STING通路激活有关。
Objective To investigate the effect of miR-488-3p on renal tubular epithelial cell injury by regulating the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon gene(STING)signaling pathway.Methods Human renal tubular epithelial HK-2 cells were divided into control,hypoxia-reoxygenation(HR),inhibition control,miR-488-3p inhibition,cGAS-STING pathway activator(RocA),and miR-488-3p inhibition+RocA groups.Quantitative real-time polymerase chain reaction was used to detect miR-488-3p expression in HK-2 cells.Cell viability was assessed using the CCK-8 assay.Enzyme-linked immunosorbent assay was used to measure the levels of interleukin(IL)-10,IL-1β,and tumor necrosis factor-α(TNF-α)in the culture supernatant of HK-2 cells.The DCFH-DA method was applied to detect the average fluorescence intensity of reactive oxygen species(ROS)in cells.Results Compared with the control group,the HR group showed increased miR-488-3p expression,IL-1βand TNF-αlevels,average fluorescence intensity of ROS,malondialdehyde(MDA)levels,apoptosis rate,and protein expression of cleaved caspase-3,Bax,cGAS,and STING,whereas cell viability,IL-10 levels,and superoxide dismutase(SOD)activity were reduced in the HR group(P<0.05).Compared with the HR and inhibition control groups,the miR-488-3p inhibition group exhibited decreased expression of miR-488-3p,IL-1βand TNF-αlevels,average fluorescence intensity of ROS,MDA levels,apoptosis rate,and protein levels of cleaved caspase-3,Bax,cGAS,and STING,whereas cell viability,IL-10 levels,and SOD activity were increased(P<0.05).Conclusion Downregulation of miR-488-3p expression mitigates HR-induced inflammation,oxidative stress,and apoptosis in HK-2 cells,thereby reducing cell damage.The underlying mechanisms may be associated with an inhibition of the cGAS-STING signaling pathway.
作者
徐芳丽
王焕
徐可
XU Fangli;WANG Huan;XU Ke(Department of Nephrology,Xinxiang Central Hospital,Xinxiang 453000,China)
出处
《中国医科大学学报》
北大核心
2025年第11期982-987,共6页
Journal of China Medical University
基金
河南省医学科技攻关计划(LHGJ20220988)。
