祛风宣肺方对豚鼠咳嗽敏感性增高模型的肺组织病理及磷酸化p38丝裂原活化蛋白激酶的影响

The effect of Qufeng Xuanfei formula on lung tissue pathology and the protein expression of phosphorylated p38 mitogen-activated protein kinase in cough sensitivity increases guinea pigs model

目的观察祛风宣肺方对豚鼠咳嗽敏感性增高模型的肺组织病理及磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)的影响。方法将40只雄性SPF级健康豚鼠随机分为空白对照组、模型组、祛风宣肺方组、西药对照组4组,每组10只。采用卵蛋白和氢氧化铝腹腔注射及卵蛋白溶液雾化的方法建立豚鼠咳嗽敏感性增高模型,造模成功后连续灌胃给药10 d,取材留取肺组织,应用电镜和光镜观察各组肺组织病理变化,应用免疫组化方法观察肺组织p-p38MAPK的表达情况。结果光镜下,空白对照组结构基本正常,模型组可见支气管管腔狭窄、上皮细胞脱落、黏膜皱襞增厚,且可见支气管黏膜层及黏膜下层、肺泡腔内及周围等炎症细胞浸润,其中以单核细胞、嗜酸性粒细胞为主。病理变化程度由重到轻依次为模型组、西药对照组、祛风宣肺方组。电镜下,空白对照组结构基本正常,模型组可见肺毛细血管增厚、基底膜增厚、肺泡炎症细胞浸润及Ⅱ型肺泡上皮细胞结构改变。病理变化程度由重到轻依次为模型组、西药对照组、祛风宣肺方组。肺组织pp38MAPK的分布情况,光镜下4组均有p-p38MAPK的阳性表达,主要表达于支气管、肺泡、肺毛细血管周围,阳性表达程度由强到弱依次为模型组、西药对照组、祛风宣肺方组、空白对照组。各组肺组织p-p38MAPK平均积分光密度数值比较,模型组、祛风宣肺方组、西药对照组较空白对照组均升高,差异均有统计学意义(P<0.05),提示咳嗽敏感性增高动物模型造模成功;祛风宣肺方组、西药对照组较模型组均降低,差异均有统计学意义(P<0.05);祛风宣肺方组较西药对照组降低,差异有统计学意义(P<0.05)。结论祛风宣肺方对豚鼠咳嗽敏感性增高模型有较好的治疗作用,其作用机制可能与修复气道损伤、抑制气道炎症细胞分泌及影响p-p38MAPK的表达水平等有关,从而减轻气道神经源性炎症,进而降低了咳嗽敏感性。

Objective To observe the effect of Qufeng Xuanfei formula on lung tissue pathology and the protein expression of phosphorylated p38 mitogen-activated protein kinase in cough sensitivity increases guinea pigs model. Methods 40 male SPF healthy guinea pigs were randomly divided into the blank control group,the model group,Qufeng Xuanfei formula group and the control group,with 10 rats in each group. The model of guinea pig with increased cough reflex sensitivity were established successfully with injecting of egg albumin and aluminium hydroxide and atomizing egg albumin,after intragastrical administration for 10 days,the lung tissue pathology under the light microscope and electron microscope and the p-p38 MAPK expression of lung tissue by immunohistochemical method were detected. Results Under the light microscopy,the blank group had normal structure,the model group could be found pathological changes in bronchostenosis,desquamation of epithelial cells,thickening of bronchial mucosa duplicature,the condition of inflammatory cell infiltration in bronchial mucosa,submucosa layer,the alveolar cavity and the surrounding,etc. The order of pathological changes from heavy to light is the model group,the western medicine control group and the Qufeng Xuanfei formula group. Under electron microscopy,the blank group had normal structure,the model group could be found pathological changes in thickening of pulmonary capillary,thickening of basement membrane,inflammatory cell infiltration in alveolar and the structure changes of alveolar type II cell,etc. The order of pathological changes from heavy to light is the model group,the western medicine control group and the Qufeng Xuanfei formula group. In the pp38 MAPK expression of lung tissue,the positive expression of p-p38 MAPK was visible in four groups,mainly expressing around bronchus,alveolar and pulmonary capillaries. The order of the positive expression level of p-p38 MAPK from strong to weak was the model group,the western medicine control group,the Qufeng Xuanfei formula group and the blank control group. By calculating p-p38 MAPK average IOD value in each group,the model group,Qufeng Xuanfei formula group,western medicine control group increased compared with the blank control group,with statistical significance（ P 〈0. 05）,proving that the animal model of increased cough reflex sensitivity could be established successfully. The qufeng xuanfei formula group and western medicine control group compared with model group respectively were lower,with statistical significance（ P 〈0. 05）; The Qufeng Xuanfei formula group compared with western medicine control group were lower,with statistical significance（ P 〈0. 05）. Conclusion Qufeng Xuanfei formula has a satisfactory curative effect on guinea pigs with increased cough reflex sensitivity. Its mechanism may be related to repair damaged airway,inhibit the secretion of airway inflammatory cells,and influence the protein expression level of p-p38 MAPK,then it could inhibit airway neurogenic inflammation and reduce increased cough reflex sensitivity.