固本防哮饮对哮喘小鼠NLRP3和TLR4的影响

Effects of Guben Fangxiao Decoction on NLRP3 and TLR4 of Asthmatic Mice

目的观察中药复方固本防哮饮对哮喘小鼠肺组织中炎症小体3(NLRP3)及Toll样受体4(TLR4)的影响并探讨其防治哮喘机制。方法采用卵蛋白(OVA)联合人呼吸道合胞病毒(RSV)致敏激发雌性Balb/c小鼠,建立哮喘持续期小鼠模型,随机分为正常组、模型组、中药组(固本防哮饮组)。苏木精-伊红染色法观察小鼠肺组织病理变化,Western blotting法及real-time q PCR法测定小鼠肺组织中NLRP3、TLR4、白细胞介素-1β(IL-1β)蛋白及mRNA表达,ELISA法测定小鼠肺组织、血清中IL-1(IL-1)及IL-18(IL-18)表达水平。结果模型小鼠肺组织内TLR4、IL-1β蛋白及mRNA表达均高于正常组(P<0.05),固本防哮饮组小鼠肺组织中TLR4、IL-1β蛋白、mRNA表达均低于模型组(P<0.05);肺组织中NLRP3蛋白及m RNA表达、肺组织及血清中IL-18蛋白表达3组间差异无统计学意义(P>0.05)。结论TLR4的激活可能与哮喘持续期气道炎症持续存在有关。固本防哮饮可能通过下调TLR4,减少IL-1的释放,减轻气道炎症,从而缓解哮喘症状。

Objective:To study the effects of Guben Fangxiao Decoction on NLRP3 and TLR4 of asthmatic mice.Methods:Female Balb/c mice were challenged with OVA and RSV to establish a persistent mouse model of asthma.They were randomly divided into the normal group,the model group and Chinese medicine group(Guben Fangxiao Decoction group).Hematoxylin-eosin staining was used to observe the pathological changes of lung tissue in mice.The expression of NLRP3,TLR4,IL-1βprotein and m RNA in lung tissue of mice was determined with Western blotting and real-time qPCR.Expression levels of IL-1βand IL-18 in mouse lung tissue and serum were detected with ELISA.Results:The expressions of TLR4,IL-1βprotein and mRNA in lung tissue of the model group were higher than those of the normal group(P<0.05).Above indices of Guben Fangxiao Decoction group were lower than those of the model group(P<0.05).There was no statistical difference in NLRP3 and IL-18 protein and mRNA expression(P>0.05).Conclusion:The activation of TLR4 may be related to the persistence of airway inflammation during asthma persistence.Guben Fangxiao Decoction may alleviate asthma symptoms by lowering TLR4,reducing IL-1βrelease and alleviating airway inflammation.