1,25-二羟维生素D3通过NLRP3/caspase-1/GSDMD信号轴干预中性粒细胞性哮喘  

1,25-dihydroxyvitamin D3 ameliorates neutrophilic asthma by regulating NLRP3/caspase-1/GSDMD signal axis

作　　者：张景鸿[1] 韦丽莹 陈一平 ZHANG Jing-hong;WEI Li-ying;CHEN Yi-ping(Dept of Emergency,the Second Affiliated Hospital of Guangxi Medical University,Nanning 530007,China;Dept of General Practice,the Affiliated Minzu Hospital of Guangxi Medical University,Nanning 530001,China)

机构地区：[1]广西医科大学第二附属医院急诊科,广西南宁530007 [2]广西医科大学附属民族医院全科医学科,广西南宁530001

出　　处：《中国药理学通报》2022年第9期1327-1333,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81860005);广西自治区自然科学基金面上项目(No 2019GXNSFAA185037,2020GXNSFAA297012)。

摘　　要：目的探讨1,25-二羟维生素D3对中性粒细胞性哮喘小鼠模型的气道炎症的干预作用,以及对NLRP3/caspase-1/GSDMD信号轴的影响。方法选取24只♀SPF级BALB/c小鼠,随机(随机数字表法)分为3组:正常对照组(A)、模型组(B)、治疗组(C),每组8只。用鸡卵清蛋白(OVA)结合脂多糖诱导的方法制作小鼠中性粒细胞性哮喘动物模型。C组在每次卵蛋白激发前给予0.1%的1,25-二羟维生素D腹腔注射(4μg·kg^(-1))。进行无创气道反应性检测,收集肺组织标本和支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF);病理HE染色、AB-PAS染色观察肺组织病理改变和气道黏液分泌情况;BALF中炎症细胞计数及分类,检测BALF中IL-1β、IL-18、IL-17水平;Ly-6G免疫组化染色了解肺组织中性粒细胞浸润;Western blot测定肺组织NLRP3、caspase-1、GSDMD蛋白表达水平。结果治疗组BALF中IL-1β、IL-18、IL-17分泌较模型组减少(P<0.05),中性粒细胞比例低于模型组,肺组织病理提示气道炎症病变较模型组减轻,AHR明显降低;肺组织NLRP3、caspase-1、GSDMD蛋白表达水平较模型组明显降低(P<0.05)。结论1,25-二羟维生素D3能减轻中性粒细胞性哮喘小鼠气道炎症和炎性细胞因子分泌,抑制NLRP3、caspase-1、GSDMD蛋白表达。1,25-二羟维生素D3可能通过调节NLRP3/caspase-1/GSDMD信号轴改善中性粒细胞性哮喘。Aim To investigate the effect of 1,25-dihydroxyvitamin D3 on airway inflammation in a mouse model of neutrophilic asthma and its influence on the NLRP3/caspase-1/GSDMD signal axis.Methods Twenty-four female SPF BALB/c mice were selected and randomly(random number table method)divided into three groups:normal control group(A),model group(B),and intervention treatment group(C),with eight mice in each group.An animal model of mouse neutrophilic asthma was made by the method induced by chicken ovalbumin(OVA)combined with lipopolysaccharide(LPS).Group C was given 0.1%1,25-(OH)Dintraperitoneal injection(4μg·kg^(-1))before each OVA challenge.A noninvasive pulmonary function test was performed to assess airway responsiveness,and lung tissues and bronchoalveolar lavage fluid(BALF)were collected.HE staining and AB-PAS staining of lung tissues were performed to observe the pathological changes and airway mucus secretion.Total inflammatory cell count and classification were carried out,and the levels of IL-1β,IL-18,and IL-17 in BALF were detected.Immunohistochemistry staining of Ly-6 G was done to confirm the presence of neutrophils in lung.The protein expressions of NLRP3,caspase-1,and GSDMD in lung tissues were determined by Western blot.Results The levels of IL-1β,IL-18,IL-17 in BALF in treatment group was lower than that in model group(P<0.05),and the proportion of neutrophils in BALF was lower than that in model group.Lung histology suggested that airway inflammation in treatment group was less than that in model group.In treatment group,AHR was significantly reduced;NLRP3,caspase-1,GSDMD protein expression levels were significantly lower than those in model group(P<0.05).Conclusions 1,25-dihydroxyvitamin D3 reduces airway inflammation and inflammatory cytokine secretion in mice with neutrophil asthma,and inhibits the expression of NLRP3,caspase-1 and GSDMD in lung.1,25-dihydroxyvitamin D3 may improve neutrophilic asthma by regulating the NLRP3/caspase-1/GSDMD signal axis.

关 键 词：哮喘 中性粒细胞 1 25-二羟维生素D3 NLRP3 CASPASE-1 GSDMD 

分 类 号：R-332[医药卫生] R364.5R331.123R562.25R977.24