ROS/TXNIP/NLRP3信号通路在高糖诱导人胚胎滋养层细胞焦亡过程中的作用  

Role of ROS/TXNIP/NLRP3 signaling pathway in high glucose-induced pyroptosis of human embryonic trophoblast cells

作　　者：时艳梅[1] 张素芹 赵明瑜[1] Shi Yanmei;Zhang Suqin;Zhao Mingyu(Department of Gynecology and Obstetrics,Yantaishan Hospital,Yantai 264003,China)

机构地区：[1]烟台市烟台山医院产科,烟台264003

出　　处：《中华内分泌外科杂志》2023年第3期312-316,共5页Chinese Journal of Endocrine Surgery

基　　金：烟台市科技计划项目(2020MSGY088)。

摘　　要：目的探究活性氧(ROS)/硫氧还蛋白相互作用蛋白(TXNIP)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路在高糖诱导下人胚胎滋养层细胞焦亡变化情况。方法体外培养人胚胎滋养层细胞,建立高糖损伤模型,分为对照组、高糖(HG)组、HG+ROS抑制剂N-乙酰-L-半胱氨酸(HG+NAC)组。噻唑蓝(MTT)法检测各组细胞存活率变化情况;二氢乙啶-ROS荧光探针法检测各组细胞的ROS水平;实时荧光定量PCR(RT-qPCR)法检测转染后各组细胞TXNIP和NLRP3 mRNA表达情况;蛋白免疫印迹分析法检测各组细胞TXNIP、NLRP3、半胱氨酸天冬氨酸蛋白酶(Caspase)-1和白细胞介素(IL)-1β及肿瘤坏死因子-α(TNF-α)、GSDMD蛋白表达水平;流式细胞术检测各组细胞焦亡情况。结果高糖诱导人胚胎滋养层细胞损伤的最佳D-葡萄糖浓度为30 mmol/L;与对照组(96.27±3.10)%相比,HG组(55.44±2.15)%人胚胎滋养层细胞存活率显著降低(P<0.05),7’二氯荧光素(DCF)荧光强度(ROS水平)、TXNIP、NLRP3蛋白表达水平、细胞焦亡数目、Caspase-1、GSDMD、IL-1β和TNF-α蛋白表达水平显著升高(P<0.05);与HG组相比,HG+NAC组(84.75±2.33)%人胚胎滋养层细胞存活率显著升高(P<0.05),DCF荧光强度(ROS水平)、TXNIP、NLRP3蛋白表达水平、细胞焦亡数目、Caspase-1、GSDMD、IL-1β和TNF-α蛋白表达水平显著降低(P<0.05)。结论抑制高糖诱导下人胚胎滋养层细胞内ROS水平,可能通过抑制TXNIP/NLRP3信号通路,促进细胞增殖,并减少焦亡的发生。Objective To investigate the changes of ROS/TXNIP/NLRP3 signaling pathway in pyroptosis of human embryonic trophoblast cells induced by high glucose.Methods Human embryonic trophoblast cells were cultured in vitro to establish high glucose injury model,and they were randomly divided into control group,high glucose(HG)group and HG+ROS inhibitor N-acetyl-L-cysteine(HG+NAC)group.MTT assay was used to detect the cell survival rate.The level of ROS in each group was detected by dihydroethidine ROS fluorescence probe.Expression of TXNIP and NLRP3 mRNA was detected by real-time quantitative PCR(RT-qPCR).Western blot analysis was used to detect the expression levels of TXNIP,NLRP3,Caspase-1,interleukin(IL)-1β,tumor necrosis factor-α(TNF-α)and GSDMD proteins.In addition,pyroptosis was detected by flow cytometry.Results The optimal glucose concentration for high glucose-induced injury of human embryonic trophoblast cells was 30 mmol/L.Compared with the control group(96.27±3.10)%,the survival rate of human embryonic trophoblast cells in HG group(55.44±2.15)%was significantly lower(P<0.05),while the fluorescence intensity(ROS level)of 7'dichlorofluorescein(DCF),the expression levels of TXNIP and NLRP3 proteins,the number of pyroptosis,expression levels of Caspase-1,GSDMD,IL-1βand TNF-αproteins were significantly higher(P<0.05);Compared with HG group,the survival rate of human embryonic trophoblast cells in HG+NAC group(84.75±2.33)%was significantly higher(P<0.05),the fluorescence intensity(ROS level)of DCF,the expression levels of TXNIP and NLRP3 proteins,the number of pyroptosis,and expression levels of Caspase-1,GSDMD,IL-1βand TNF-αproteins were significantly lower(P<0.05).Conclusion Inhibition of ROS level in human embryonic trophoblast cells induced by high glucose may promote cell proliferation and reduce the occurrence of pyroptosis by inhibiting TXNIP/NLRP3 signaling pathway.

关 键 词：人胚胎滋养层细胞 高糖 焦亡 活性氧/硫氧还蛋白相互作用蛋白/NOD样受体蛋白-3通路 

分 类 号：R711[医药卫生—妇产科学]