摘要
目的探讨NLRP3/SDF-1信号通路在哮喘气道炎症过程的作用机制。方法36只4周龄雄性BALB/c小鼠随机分为对照组、哮喘组和布地奈德(BUD)组,每组12只。采用腹腔注射卵清蛋白(OVA)构建小鼠哮喘模型,H-E染色观察肺组织病变,计数肺泡灌洗液(BALF)中炎性细胞的数量,Western-blot检测肺组织NLRP3、SDF-1和CXCR4蛋白的表达水平;采用慢病毒构建BEAS-2B/sh NLRP3细胞系,CCK-8检测细胞增殖活力,使用不同浓度(0、200、400、800 U/mL)的屋尘螨(HDM)干预细胞,模拟体外哮喘模型,比较单独使用NLRP3抑制剂(MCC950)和联合BUD治疗时,NLRP3、p-NF-κB和SDF-1蛋白的表达水平。结果(1)炎性细胞数:哮喘组较对照组增加(P<0.001),而BUD组较哮喘组降低(P<0.001)。(2)小鼠肺组织的NLRP3、SDF-1和CXCR4蛋白的表达水平:哮喘组3个指标均较对照组升高(P<0.001、P<0.01和P<0.001),BUD组则均较哮喘组降低(均为P<0.001)。(3)细胞的NLRP3、p-NF-κB和SDF-1蛋白的表达水平:400、800 U/mL HDM组的p-NF-κB表达水平均较对照组升高(P<0.05和P<0.01)。经HDM干预后,BEAS-2B/sh NLRP3组细胞的p-NF-κB水平降低(P<0.01、P<0.01和P<0.001),且SDF-1蛋白的变化趋势同p-NF-κB蛋白。与单独使用MCC950组比较,联合使用MCC950和BUD治疗组的3个指标的表达水平均降低(P<0.05、P<0.05和P<0.01)。结论BUD可通过限制NLRP3/SDF-1信号抑制哮喘的发生,联合使用BUD和NLRP3抑制剂可能成为哮喘的治疗策略。
Objective To explore the mechanism of NLRP3/SDF-1 signaling in the inflammatory process of asthmatic airways.Methods Thirty-six 4-week-old male BALB/c mice were randomly divided into control group,asthma group and budesonide(BUD)group,with 12 mice in each group.A mouse asthma model was constructed by intraperitoneal injection of ovalbumin(OVA).H-E staining was used to observe lung tissue lesions,and the number of inflammatory cells in alveolar lavage fluid(BALF)was counted.Western-blot was used to detect the protein expression levels of NLRP3,SDF-1 and CXCR4 in lung tissue.BEAS-2B/sh NLRP3 cell line was constructed by using lentivirus,CCK-8 was used to detect cell proliferation activity.Different concentrations(0,200,400,800 U/mL)of house dust mite(HDM)were used to treat cells and simulate an in vitro asthma model.The NLRP3,p-NF-κB and SDF-1 protein expression levels were compared in NLRP3 inhibitor(MCC950)alone and in combination with BUD treatment.Results(1)Inflammatory cell counts:increased in the asthma group compared to the control group(P<0.001),while decreased in the BUD group compared to the asthma group(P<0.001).(2)Protein expression levels of NLRP3,SDF-1 and CXCR4 in mouse lung tissue:the asthma group showed an increase in all three indicators compared to the control group(P<0.001,P<0.01 and P<0.001),while the BUD group showed a decrease compared to the asthma group(P<0.001).(3)Protein expression levels of NLRP3,p-NF-κB and SDF-1 in cells:The p-NF-κB expression levels in HDM(400 U/mL)and HDM(800 U/mL)groups were higher than that in the control group(P<0.05,P<0.01).After HDM intervention,the p-NF-κB level of cells in BEAS-2B/sh NLRP3 group decreased(P<0.01,P<0.01 and P<0.001),and the SDF-1 protein had the same trend as the p-NF-κB protein.Compared with the group treated with MCC950 alone,the combined treatment of MCC950 and BUD resulted in a decrease in protein expression of all three indicators(P<0.05,P<0.05 and P<0.01).Conclusion BUD can inhibit asthma by limiting the NLRP3/SDF-1 signaling,and the
作者
李玉娟
叶建芬
汪杨
王玲
徐晓燕
游爱萍
LI Yujuan;YE Jianfen;WANG Yang;WANG Ling;XU Xiaoyan;YOU Aiping(Department of Pediatrics,Fujian Medical University Affiliated Nanping First Hospital,Nanping 353000,China;Fujian Maternity and Child Health Hospital,College of Clinical Medicine for Obstetrics&Gynecology and Pediatrics,Fujian Medical University,Fuzhou 350005,China)
出处
《福建医科大学学报》
2024年第1期20-25,共6页
Journal of Fujian Medical University
基金
福建省自然科学基金项目(2021J011431)。
