摘要
目的观察细颗粒物(PM_(2.5))暴露对氧糖剥夺后再复氧复糖(OGD/R)大鼠神经小胶质细胞系GMI-R1的损伤作用,并基于硫氧还蛋白相互作用蛋白(TXNIP)/NLR家族Pyrin结构域3(NLRP3)通路探讨相关机制。方法培养GMI-R1细胞并分为对照组、模型组、实验组、TXNIP抑制剂组。模型组、实验组、TXNIP抑制剂组制作OGD/R模型,对照组常规培养。实验组和TXNIP抑制剂组在造模前先进行50μg/m L PM_(2.5)暴露24 h,TXNIP抑制剂组PM_(2.5)暴露前给予10μmol/L的TXNIP抑制剂鲁斯可皂苷元预处理30 min。复氧24 h后采用流式细胞术检测死亡细胞,采用ELISA法检测各组培养基上清中的白细胞介素(IL)-18和IL-1β,采用Western blotting法检测各组细胞中的TXNIP、NLRP3、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白水解酶1(Caspase-1)、消皮素D(GSDMD),采用免疫荧光法检测GSDMD-N。结果对照组、模型组、实验组细胞死亡率及培养液上清中IL-18、IL-1β水平依次升高,TXNIP抑制剂组细胞死亡率及培养液上清中IL-18、IL-1β水平低于实验组(P均<0.01)。对照组、模型组、实验组细胞中TXNIP、NLRP3、ASC、Caspase-1蛋白及焦亡相关蛋白GSDMD、GSDMD-N表达依次增高,TXNIP抑制剂组NLRP3、ASC、Caspase-1、GSDMD、GSDMD-N蛋白表达低于实验组(P均<0.05)。结论PM_(2.5)暴露能加重OGD/R下GMI-R1细胞的损伤,加重炎症反应,机制可能与促进TXNIP/NLRP3通路活化和细胞焦亡有关。
Objective To investigate the impact of PM_(2.5) exposure on microglia injury in rats(GMI-R1)subjected to oxygen-glucose deprivation and recovery(OGD/R),and to explore the related mechanism based on the thioredoxin-interacting protein(TXNIP)/NOD-like receptor family pyrin domain-containing 3(NLRP3)pathway.Methods GMI-R1 cells were cultured and divided into the control group,model group,experimental group,and TXNIP inhibitor group,respectively.OGD/R models were established in the model group,experimental group and TXNIP inhibitor group,and cells in the control group were cultured routinely.Cells in the experimental group and TXNIP inhibitor group were exposed to PM_(2.5)(50μg/mL)for 24 h before modeling.Cells in the TXNIP inhibitor group were pretreated with 10μmol/L ruscogenin(TXNIP inhibitor)for 30 minutes before PM_(2.5) exposure.After 24 h reoxygenation,flow cytometry was used to detect dead cells,and ELISA was used to detect interleukin(IL)-18 and IL-1βin the culture medium supernatant of each group.Western blotting was used to detect TXNIP,NLRP3,apoptosis-associated speck-like protein(ASC),Caspase-1,and Gasdermin D(GSDMD)in cells of each group,and immunofluorescence was used to detect GSDMD-N.Results The cell death rate and the levels of IL-18 and IL-1βin the culture supernatant of the control group,the model group and the experimental group increased in turn,and the cell death rate and the levels of IL-18 and IL-1βin the culture supernatant of the TXNIP inhibitor group were lower than those of the experimental group(all P<0.01).The expression levels of TXNIP,NLRP3,ASC,Caspase-1 and pyroptosis-related proteins GSDMD and GSDMD-N in the control group,model group and experimental group increased in turn;the expression levels of NLRP3,ASC,Caspase-1 and pyroptosis-related proteins GSDMD and GSDMD-N in the TXNIP inhibitor group were lower than those in the experimental group(all P<0.05).Conclusion PM_(2.5) exposure can aggravate the injury of GMI-R1 cells and intensify the inflammatory response under OGD/R,and the mechanism may be related to promoting the activation of TXNIP/NLRP3 pathway and pyroptosis.
作者
周宇
李伟
马勇
李斌
柴尔青
ZHOU Yu;LI Wei;MA Yong;LI Bin;CHAI Erqing(不详;First Clinical Medical College,Lanzhou University,Lanzhou 730000,China)
出处
《山东医药》
CAS
2024年第8期28-32,共5页
Shandong Medical Journal
基金
甘肃省自然科学基金资助项目(20JR10RA379,23JRRA1308)。
