杨梅素调节NLRP3/Caspase-1/IL-1β信号通路对肺炎链球菌诱导肺泡上皮细胞焦亡的影响  

The Effect of Myricetin on Streptococcus Pneumoniae-Induced Pyroptosis of Alveolar Epithelial Cells by Modulating NLRP3/Caspase-1/IL-1β Signaling Pathway

作　　者：王永梅 蒋燕 黄嘉阳 王香红 WANG Yongmei;JIANG Yan;HUANG Jiayang;WANG Xianghong(Department of Pediatrics,Suzhou High Tech Zone People's Hospital,Suzhou 215129,China)

机构地区：[1]苏州高新区人民医院儿科,江苏苏州215129

出　　处：《标记免疫分析与临床》2024年第9期1704-1710,共7页Labeled Immunoassays and Clinical Medicine

摘　　要：目的探讨杨梅素(myricetin,MYR)调节NOD样受体热蛋白结构域相关蛋白3(nucleotide-binding oligomerization domain,leucine-rich repeat and pyrin domain-containing 3,NLRP3)/半胱氨酸蛋白酶-1(Caspase-1)/白细胞介素-1β(IL-1β)信号通路对肺炎链球菌诱导肺泡上皮细胞焦亡的影响。方法将人II型肺泡上皮细胞(human type II alveolar epithelial cells,AT II)随机分为5组:对照组、模型组、L-MYR组(20μg/mL)、M-MYR组(40μg/mL)、H-MYR组(80μg/mL)、ATP组(80μg/mL MYR+2 mmol/L NLRP3/Caspase-1/IL-1β信号通路激活剂ATP),且除对照组正常培养,其余各组以肺炎链球菌感染建立损伤模型。采用MTT和平板克隆实验检测细胞增殖;TUNEL检测细胞焦亡;ELISA检测细胞中白细胞介素-1β(IL-1β)、白细胞介素18(IL-18)、乳酸脱氢酶(lactic dehydrogenase,LDH)水平;Western blotting检测细胞中NLRP3、cleaved caspase-1、焦孔素D(gasdermin D,GSDMD)、IL-1β蛋白表达。结果与对照组相比,模型组细胞的A 490、克隆数降低,细胞焦亡率、IL-1β、IL-18、LDH、NLRP3、cleaved caspase-1、GSDMD、IL-1β表达升高(P<0.05);与模型组相比,L-MYR组、M-MYR组、H-MYR组细胞的A 490、克隆数升高,细胞焦亡率、IL-1β、IL-18、LDH、NLRP3、cleaved caspase-1、GSDMD、IL-1β表达较低(P<0.05);与H-MYR组相比,ATP组细胞的A 490、克隆数降低,细胞焦亡率、IL-1β、IL-18、LDH、NLRP3、cleaved caspase-1、GSDMD、IL-1β表达升高(P<0.05)。结论MYR可能通过抑制NLRP3/Caspase-1/IL-1β信号通路,减轻肺炎链球菌诱导的肺泡上皮细胞炎症反应,抑制细胞焦亡。Objective To investigate the effect of myricetin(MYR)on alveolar epithelial cell pyroptosis induced by Streptococcus pneumoniae through modulating NOD-like receptor thermal protein domain associated protein 3(NLRP3)/cysteine protease-1(Caspase-1)/interleukin-1β(IL-1β)signaling pathway.Methods Human type II alveolar epithelial cells(AT II.)were randomly separated into five groups:control group,model group,L-MYR group(20μg/mL),M-MYR group(40μg/mL),H-MYR group(80μg/mL),and ATP group(80μg/mL MYR+2 mmol/L NLRP3/Caspase-1/IL-1βsignaling pathway activator ATP).Except for applying the normal culture condition in the control group,the damage model was established with Streptococcus pneumoniae infection in all the other groups.MTT and plate cloning method were used to detect the proliferation of cells.TUNEL was used to detect pyroptosis of cells.ELISA was applied to detect the levels of interleukin-1β(IL-1β),interleukin-18(IL-18),and lactate dehydrogenase(LDH).Western blot was used to detect the protein expressions of NLRP3,cleaved caspase-1,pyocorin D(GSDMD)and IL-1β.Results Compared with the control group,A 490 and the number of clones in the model group decreased,while the cell pyroptosis rate,and the expressions of IL-1β,IL-18,LDH,NLRP3,cleaved caspase-1,GSDMD and IL-1βincreased(P<0.05).Compared with the model group,A 490 and the number of clones in the L-MYR group,M-MYR group and H-MYR group increased,while the cell pyroptosis rate,and the expressions of IL-1β,IL-18,LDH,NLRP3,cleaved caspase-1,GSDMD and IL-1βdecreased(P<0.05).Compared with the H-MYR group,A 490 and the number of clones in the ATP group decreased,while the cell pyroptosis rate,and the expressions of IL-1β,IL-18,LDH,NLRP3,cleaved caspase-1,GSDMD and IL-1βincreased(P<0.05).Conclusion MYR could alleviate the inflammatory response of alveolar epithelial cells induced by Streptococcus pneumoniae through inhibiting the NLRP3/Caspase-1/IL-1βsignaling pathway,thereby inhibiting cell pyroptosis.

关 键 词：杨梅素 NOD样受体热蛋白结构域相关蛋白3/半胱氨酸蛋白酶-1/白细胞介素-1β信号通路 肺炎链球菌感染 细胞焦亡 

分 类 号：Q255[生物学—细胞生物学]