NOD样受体热蛋白结构域相关蛋白3介导的细胞焦亡对哮喘大鼠炎症水平的调控作用

Regulation of pyroptosis mediated by NOD-like receptor pyrin domain-related protein 3 on inflammatory in asthmatic rats

目的:探讨NOD样受体热蛋白结构域相关蛋白3(NLRP3)在哮喘中炎症水平的调控作用机制。方法:从GSE40732和GSE69683数据集中分析哮喘组和对照组之间的差异表达基因(DEGs),并鉴定程序性细胞死亡在哮喘中的水平。在NLRP3敲降大鼠中建立哮喘大鼠模型,通过HE染色和Tunel染色分析肺组织的病理变化和凋亡水平,通过免疫组化检测肺组织中NLRP3的表达,通过实时荧光定量聚合酶链反应(RT-qPCR)和Western blot检测NLRP3介导细胞焦亡相关mRNA和蛋白表达的改变。结果:哮喘组和对照组之间鉴定了926个差异表达基因(DEGs),细胞焦亡在哮喘中的水平显著高于对照组。哮喘模型中的炎症、凋亡和NLRP3水平明显高于对照组,而在NLRP3敲降后,哮喘大鼠中的炎症和凋亡水平降低。与对照组比较,NLRP3、Gasdermin D蛋白(GSDMD)、胱天蛋白酶-1(Caspase-1)和Caspase-8在哮喘大鼠模型中的表达升高,高迁移率族蛋白1(HMGB1)的表达降低(均P<0.05),这些异常表达在NLRP3敲降后得到了显著的改善(P<0.05)。结论:NLRP3通过细胞焦亡信号可能在哮喘中发挥促炎促凋亡作用,阻断该通路可能是改善哮喘炎症的潜在治疗策略。

Objective:To explore the regulatory mechanism of NOD-like receptor pyrin domain-related protein 3(NLRP3)in asthma inflammation.Methods:Differential expression genes(DEGs)between asthma and control groups were analyzed from the GSE40732 and GSE69683 datasets,and the levels of programmed cell death were identified.An asthma rat model was established in NLRP3 knockdown rats,and pathological changes and apoptosis levels in lung tissue were analyzed using HE staining and TUNEL staining.Immunohistochemistry was employed to detect NLRP3 expression in lung tissue,and changes in NLRP3-mediated pyroptosis-related mRNA and protein expression were measured by RT-qPCR and Western blot.Results:A total of 926 DEGs were identified between asthma and control groups,and the level of pyroptosis was significantly higher in asthma(P<0.05).In the asthma model,inflammation,apoptosis,and NLRP3 levels were significantly higher compared to the control group(all P<0.05).However,after NLRP3 knockdown,inflammation and apoptosis levels were reduced in asthma rats(all P<0.05).Compared to the control group,the expressions of NLRP3,GSDMD,Caspase-1,and Caspase-8 were upregulated,while HMGB1 expression was downregulated in the asthma rat model(all P<0.05).These abnormal expressions were significantly improved after NLRP3 knockdown(all P<0.05).Conclusion:NLRP3 may exert a pro-inflammatory and pro-apoptotic role in asthma through the pyroptosis signaling pathway,and blocking this pathway can be a potential therapeutic strategy to improve asthma inflammation.