基于IL-4/JAK1/STAT6信号通路影响巨噬细胞M2极化探讨培元定喘汤治疗哮喘气道炎症的作用机制

Mechanism of Peiyuan Dingchuan Decoction in treating asthma airway inflammation based on the influence of IL-4/JAK1/STAT6 signal pathway on macrophage M2 polarization

目的:观察培元定喘汤对哮喘小鼠气道炎症及JAK1/STAT6信号通路的影响,探讨培元定喘汤对哮喘气道炎症的作用机制。方法:40只C57BL/6小鼠随机分为4组,每组10只,除空白组外,模型组、布地奈德组、培元定喘汤组应用卵清蛋白(OVA)雾化及激发的方式建立哮喘小鼠模型。观察各组小鼠的一般状态;ELISA检测肺泡灌洗液(BALF)中白细胞介素(IL)-4、IL-5、IL-13及血清中IgE水平;HE染色法观察小鼠肺组织病理形态学的变化;qRT-PCR法检测肺组织中IL-4、JAK1、STAT6、Arg-1 mRNA表达;免疫组化法检测肺组织中Arg-1和CD163的表达;免疫荧光染色检测肺组织IL-4的表达。结果:与模型组比较,布地奈德组和培元定喘汤组病理特征明显减轻;布地奈德组和培元定喘汤组BALF中IL-4、IL-5、IL-13及血清中IgE水平均显著降低(P<0.05),肺组织中IL-4、JAK1、STAT6、Arg-1 mRNA表达均显著降低(P<0.05),肺组织Arg-1及CD163表达显著降低(P<0.05),肺组织中IL-4的荧光强度明显减弱。结论:培元定喘汤可以通过抑制IL-4/JAK1/STAT6信号通路的激活抑制巨噬细胞M2极化,从而达到治疗哮喘气道炎症的作用。

Objective:To observe the effects of Peiyuan Dingchuan Decoction on airway inflammation and JAK1/STAT6 signaling pathway in asthma mice,and explore the mechanism of Peiyuan Dingchuan Decoction on airway inflammation in asthma.Methods:Forty C57BL/6 mice were randomly divided into 4 groups,with 10 mice in each group.Except for the blank group,the model group,budesonide group,and Peiyuan Dingchuan Decoction group were used to establish asthma mouse models by nebulizing and stimulating ovalbumin(OVA).The general state of each group mice was observed;the expressions of IL-4,IL-5,IL-13 in BALF and IgE levels in serum were detected by ELISA;HE staining method was used to observe the pathological and morphological changes of mouse lung tissue;The mRNA expressions of IL-4,JAK1,STAT6,Arg-1 in lung tissue were detected by qRT-PCR method;Immunohistochemical method was used to detect the expressions of Arg-1 and CD163 in lung tissue;Immunofluorescence staining was used to detect the expression of IL-4 in lung tissue.Results:Compared with the model group,the pathological characteristics of the budesonide group and the Peiyuan Dingchuan Decoction group were significantly reduced;The expressions of IL-4,IL-5,IL-13 in BALF,and IgE level in serum were significantly reduced in the budesonide group and the Peiyuan Dingchuan Decoction group showed a decrease(P<0.05).The mRNA expressions of IL-4,JAK1,STAT6 and Arg-1were significantly reduced in the budesonide group and Peiyuan Dingchuan Decoction group(P<0.05).The expressions of Arg-1and CD163 in the lung tissue of mice in the budesonide group and Peiyuan Dingchuan Decoction group were significantly reduced(P<0.05);The fluorescence intensity of IL-4 in the budesonide group and Peiyuan Dingchuan Decoction group was significantly reduced.Conclusion:Peiyuan Dingchuan Decoction can inhibit macrophage M2 polarization by inhibiting the activation of IL-4/JAK1/STAT6 signaling pathway,thereby achieving the therapeutic effect of asthma airway inflammation.