郑州大学学报（医学版）2025，Vol.60Issue(5)：603-608,6.DOI:10.13705/j.issn.1671-6825.2025.01.072
RIP3参与的线粒体DNA-cGAS-STING信号通路在HK-2细胞缺氧-复氧模型中的作用
Role of RIP3-involved mitochondrial DNA-cGAS-STING signaling path-way in hypoxia-reoxygenation model of HK-2 cells
摘要
Abstract
Aim:To explore the mechanism of RIP3-involved mitochondrial DNA(mtDNA)-cGAS-STING signaling pathway in HK-2 hypoxia-reoxygenation(H-R)cell model.Methods:The H-R cell model was constructed.HK-2 cells in logarithmic growth phase were cultured in a special medium without glucose or serum under hypoxia condition for 3,6 and 12 hours,and then reoxygenated,and control group was set up at the same time.Cell viability was detected by MTT,RIP3,cGAS,STING,TBK1,IRF3,IFN-α,IFN-β,TNF-α,IL-1β and NF-κB protein expression levels by Western blot,RIP3,cGAS,STING mRNA expression levels by RT-qPCR.Cell transfection and addition of RU.521(cGAS inhibitor),Mito-TEMPO(mitochondrial ROS-specific scavenger),and ethidium bromide(EtBr)were used to verify the cGAS-STING path-way.mtDNA level was determined by immunofluorescence staining.Results:Compared with the control group,the cell sur-vival rate of the H-R 3,6,and 12 groups decreased,and the expression levels of related proteins RIP3,cGAS,STING,TBK1,IRF3,IFN-α,IFN-β,TNF-α,IL-1β,NF-κB and the expression levels of RIP3,STING mRNA in the mtDNA-cGAS-STING signaling pathway increased,and the expression levels of cGAS mRNA in the H-R 6 and 12 groups increased(P＜0.05).Compared with the H-R 12+si-NC group,the cell survival rate of the H-R 12+si-cGAS group increased,and the levels of RIP3,cGAS,and STING mRNA decreased;compared with the H-R 12 group,the protein expression levels of RIP3,cGAS,STING,TBK1,IRF3,IFN-β,and IL-1β in the H-R 12+RU.521 group and the H-R 12+Mito-TEMPO group de-creased(P＜0.05).The treatment of H-R for 12 hours increased the cytoplasmic double-stranded DNA and co-localized with mitochondria,while EtBr treatment decreased cytoplasmic mtDNA(P＜0.05).Conclusion:H-R affects cell viability and the mtDNA-cGAS-STING signaling pathway,in which cGAS plays an important role,and cytoplasmic mtDNA has a reg-ulatory effect on the cGAS-STING axis.关键词
缺氧-复氧/HK-2细胞/RIP3/线粒体DNA-cGAS-STING信号通路Key words
hypoxia-reoxygenation/HK-2 cell/RIP3/mitochondrial DNA-cGAS-STING signaling pathway分类
医药卫生引用本文复制引用
孔亚坤,王焕,尹俊杰,熊承云,吴苏豫,张莹,赵建林..RIP3参与的线粒体DNA-cGAS-STING信号通路在HK-2细胞缺氧-复氧模型中的作用[J].郑州大学学报（医学版）,2025,60(5):603-608,6.基金项目
河南省医学科技攻关计划项目(LHGJ20220988) （LHGJ20220988）
