固本防哮饮对支气管哮喘缓解期小鼠免疫失衡的影响

Effects of Guben Fangxiao Decoction(固本防哮饮) on Immune Imbalance in Mice with Bronchial Asthma during Remission

目的探讨固本防哮饮治疗支气管哮喘缓解期的可能作用机制。方法 36只BALB/c雌性小鼠随机分为正常组、模型组、孟鲁司特钠组、固本防哮饮预防组、固本防哮饮高剂量组及固本防哮饮等效剂量组,每组6只。除正常组外其余各组采用卵蛋白致敏、雾化联合呼吸道合胞病毒滴鼻诱导建立支气管哮喘缓解期小鼠模型。造模后正常组、模型组给予蒸馏水20 ml/kg灌胃,固本防哮饮等效剂量、高剂量组分别给予24、36 g/(kg·d)固本防哮饮灌胃,孟鲁司特钠组给予孟鲁司特钠片2. 6 mg/(kg·d)灌胃,每日1次,共28天。固本防哮饮预防组从致敏第1天给予24 g/(kg·d)的固本防哮饮每日灌胃1次,共83天。比较各组小鼠肺组织病理变化,采用实时定量PCR法检测肺组织中胸腺基质淋巴细胞生成素(TSLP)、胸腺基质淋巴细胞生成素受体(TSLPR)、GATA3 mRNA表达,Western Blotting法检测TSLP、TSLPR、GATA3蛋白表达水平,酶联免疫吸附试验测定血清TSLP、TSLPR蛋白表达。结果病理学结果显示,正常组小鼠气道上皮平整,支气管管腔光滑,无明显炎性细胞浸润;模型组小鼠气道上皮不完整,肺泡壁增厚,同时气道黏膜层、黏膜下层以及血管周围组织可见大量炎性细胞浸润;固本防哮饮各治疗组和孟鲁司特钠组小鼠肺组织病理炎症均较模型组减轻,且各治疗组病理差异不明显。与正常组比较,模型组小鼠肺组织TSLP、TSLPR、GATA3 mRNA及蛋白表达显著升高(P <0. 05)。与模型组比较,各给药组小鼠肺组织TSLP、TSLPR、GATA3 mRNA及蛋白表达均不同程度降低(P <0. 05)。各组小鼠血清中TSLP、TSLPR蛋白表达差异无统计学意义(P> 0. 05)。结论固本防哮饮可改善支气管哮喘缓解期的肺组织病理改变,其机制可能是抑制肺组织TSLP、TSLPR、GATA3表达,纠正肺组织Th1/Th2的偏移,从而调控机体的免疫功能。

Objective To investigate the possible mechanism of Guben Fangxiao Decoction(固本防哮饮) in the treatment of bronchial asthma during remission. Methods A total of 36 female BALB/c mice were randomly divided into a normal group,a model group,a montelukast sodium group,a Guben Fangxiao Decoction prevention group,a Guben Fangxiao Decoction high-dose group and a Guben Fangxiao Decoction equivalent dose group,with 6 mice in each group. Except the normal group,the other groups were induced by ovalbumin sensitization,nebulization combined with respiratory syncytial virus to establish a mouse model of bronchial asthma during remission. After the model was established,the normal group and the model group were given 20 ml/kg of distilled water for intragastric administration. The equivalent dose and high dose group of Guben Fangxiao Decoction were given 24,36 g/( kg·d),respectively. In the montelukast sodium group,montelukast sodium tablets were administered 2. 6 mg/( kg·d) intragastrically once a day for 28 days. The Guben Fangxiao Decoction prevention group was given a daily dose of 24 g/( kg·d)of Guben Fangxiao Decoction on the first day of sensitization for 83 days. The pathological changes of lung tissue in mice were compared between the groups. The expression of thymic stromal lymphopoietin( TSLP),thymic stromal lymphopoietin receptor( TSLPR) and GATA3 mRNA was detected by real-time quantitative PCR. Western blotting was used to detect TSLP.,TSLPR,GATA3 protein expression levels. Enzyme-linked immunosorbent assay was used to determine serum TSLP,TSLPR protein expression. Results The pathological results showed that the airway epithelium was smooth in the normal group,the bronchial lumen was smooth,and there was no obvious inflammatory cell infiltration. In the model group,the airway epithelium was incomplete,and the alveolar wall was thickened. The airway mucosa and submucosa could see a large number of inflammatory cell infiltration in the perivascular tissue;the pathological inflammation of lung tissue in the treatment group and the montelukast sodium group were all reduced compared with the model group,and the pathological difference of each treatment group was not obvious. Compared with the normal group,the expression of TSLP,TSLPR and GATA3 mRNA and protein in the lung tissue of the model group was significantly increased( P < 0. 05). Compared with the model group,the expressions of TSLP,TSLPR and GATA3 mRNA and protein in lung tissue of each group were decreased( P < 0. 05). There was no significant difference in the expression of TSLP and TSLPR protein in serum of each group( P > 0. 05). Conclusion Guben Fangxiao Decoction can improve the pathological changes of lung tissue in the remission period of bronchial asthma.The mechanism may be to inhibit the expression of TSLP,TSLPR and GATA3 in lung tissue,correct the shift of Th1/Th2 in lung tissue,and regulate the immune function of the body.