학술논문
NLRP3炎性小体对哮喘小鼠TGF-β1/Smad3信号通路的调控及机制研究 / Regulation and mechanism of NLRP3 inflammasome on TGF-β1/Smad3 signa-ling pathway in asthmatic mice
Document Type
Academic Journal
Author
Source
遵义医科大学学报 / Journal of Zunyi Medical University. 46(12):1141-1147
Subject
Language
Chinese
ISSN
1000-2715
Abstract
目的 观察NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎性小体对哮喘小鼠转化生长因子β1(TGF-β1)/Smad3信号通路的调控作用,并探讨其对哮喘气道重塑影响的可能机制.方法 将24只雌性BALB/c小鼠随机分成正常对照组(NC组,n=8)、哮喘组(AS组,n=8)、哮喘组+MCC950干预组(AS+MCC950干预组,n=8).AS组和AS+MCC950干预组通过给予鸡卵清白蛋白致敏和雾化吸入激发,制作哮喘小鼠模型,AS+MCC950干预组予NLRP3特异性抑制剂MCC950(10 mg/kg)进行干预.采用HE和AB-PAS染色观察各组小鼠肺组织的病理形态,qRT-PCR检测肺组织中NLRP3 mRNA表达,IHC染色检测肺组织NLRP3、TGF-β1和Smad3蛋白表达,Western blot检测肺组织中NLRP3蛋白表达,ELISA法检测支气管肺泡灌洗液(BALF)中炎症因子IL-1 β和IL-18水平.结果 与NC组比较,AS组小鼠BALF中 IL-1β和IL-18表达、气道上皮杯状细胞和黏液分泌、肺组织中NLRP3、TGF-β1、Smad3蛋白表达和NLRP3 mRNA表达水平均升高(P＜0.05),使用MCC950干预后,AS+MCC950干预组小鼠上述指标的表达水平较AS组下降(P＜0.05).结论 MCC950可以特异性的抑制NLRP3炎性小体的活化,下调炎症因子IL-1β和IL-18分泌,抑制肺组织中TGF-β1和Smad3的表达,其可能机制是通过干预NLRP3/IL-1 β/TGF-β1/Smad3信号通路来抑制哮喘小鼠气道重塑,说明NLRP3炎性小体参与调控哮喘小鼠TGF-β1/Smad3信号通路.
Objective To observe the regulatory effect of NOD-like receptor protein3(NLRP3)inflammasome on transforming growth factor β1(TGF-β1)/Smad3 signaling pathway in asthmatic mice,and to explore the pos-sible mechanism of its effect on airway remodeling.Methods Twenty four female BALB/c mice were randomly divided into normal control group(NC group,n=8),asthma group(AS group,n=8),asthma+MCC950 in-tervention group(AS+MCC950 intervention group,n=8).The AS group and AS+MCC950 intervention group were sensitized by chicken ovalbumin and challenged by aerosol inhalation to make asthma mouse models,and the AS+MCC950 intervention group was intervened with NLRP3-specific inhibitor MCC950(10 mg/kg).HE and AB-PAS staining were used to observe the pathological appearance of lung tissue of mice in each group,qRT-PCR was used to detect the NLRP3 mRNA expression in lung tissue,IHC staining was used to detect lung tissue NLRP3,TGF-β1 and Smad3 protein expression,Western blot was used to detect the NLRP3 protein ex-pression in lung tissue,and ELISA was used to detect the levels of inflammatory factors IL-1 β and IL-18 in the bronchoalveolar lavage fluid(BALF)of mice.Results Compared with the NC group,the expression of 1L-1βand IL-18 in BALF,airway epithelial goblet cells and mucus secretion,and the expression of NLRP3,TGF-β1,Smad3 protein and NLRP3 mRNA in the lung tissue of mice in the AS group were all increased(P＜0.05).Af-ter MCC950 intervention,the expression levels of the above indicators in the AS+MCC950 intervention group were lower than those in the AS group(P＜0.05).Conclusion MCC950 can specifically inhibit the activation of NLRP3 inflammasome,down-regulate the secretion of inflammatory factors IL-1 β and IL-18,and inhibit the ex-pression of TGF-β1 and Smad3 in lung tissue.The possible mechanism is through the intervention of NLRP3/IL-1β/TGF-β1/Smad3 signaling pathway to inhibit airway remodeling in asthmatic mice,indicating that the NLRP3 inflammasome is involved in regulating TGF-β1/Smad3 signaling pathway in asthmatic mice.
Objective To observe the regulatory effect of NOD-like receptor protein3(NLRP3)inflammasome on transforming growth factor β1(TGF-β1)/Smad3 signaling pathway in asthmatic mice,and to explore the pos-sible mechanism of its effect on airway remodeling.Methods Twenty four female BALB/c mice were randomly divided into normal control group(NC group,n=8),asthma group(AS group,n=8),asthma+MCC950 in-tervention group(AS+MCC950 intervention group,n=8).The AS group and AS+MCC950 intervention group were sensitized by chicken ovalbumin and challenged by aerosol inhalation to make asthma mouse models,and the AS+MCC950 intervention group was intervened with NLRP3-specific inhibitor MCC950(10 mg/kg).HE and AB-PAS staining were used to observe the pathological appearance of lung tissue of mice in each group,qRT-PCR was used to detect the NLRP3 mRNA expression in lung tissue,IHC staining was used to detect lung tissue NLRP3,TGF-β1 and Smad3 protein expression,Western blot was used to detect the NLRP3 protein ex-pression in lung tissue,and ELISA was used to detect the levels of inflammatory factors IL-1 β and IL-18 in the bronchoalveolar lavage fluid(BALF)of mice.Results Compared with the NC group,the expression of 1L-1βand IL-18 in BALF,airway epithelial goblet cells and mucus secretion,and the expression of NLRP3,TGF-β1,Smad3 protein and NLRP3 mRNA in the lung tissue of mice in the AS group were all increased(P＜0.05).Af-ter MCC950 intervention,the expression levels of the above indicators in the AS+MCC950 intervention group were lower than those in the AS group(P＜0.05).Conclusion MCC950 can specifically inhibit the activation of NLRP3 inflammasome,down-regulate the secretion of inflammatory factors IL-1 β and IL-18,and inhibit the ex-pression of TGF-β1 and Smad3 in lung tissue.The possible mechanism is through the intervention of NLRP3/IL-1β/TGF-β1/Smad3 signaling pathway to inhibit airway remodeling in asthmatic mice,indicating that the NLRP3 inflammasome is involved in regulating TGF-β1/Smad3 signaling pathway in asthmatic mice.