复方银杏颗粒对肺纤维化大鼠模型的影响及作用机制研究

Effects of compound recipe Gingko granules on rat models with pulmonary fibrosis and its possible action mechanism

目的观察复方银杏颗粒对肺纤维化大鼠模型的影响,探讨复方银杏颗粒治疗肺纤维化的可能机制。方法 50只雄性SD大鼠一次性气管内注射盐酸博来霉素A5建立大鼠肺纤维化模型。将造模成功的大鼠随机模型组、阳性对照组,复方银杏颗粒高、中、低剂量组,每组10只,另选10只作为正常组,造模第2天起,正常组和模型组用0. 9%氯化钠溶液灌胃,阳性对照组10 ml/mg(0. 32 mg/ml),复方银杏颗粒高、中、低剂量组(15 g/kg、7. 5 g/kg、3. 75g/kg)灌胃给药,大鼠处死抽取肺泡灌洗液,计数白细胞并检测BALF蛋白与血清总蛋白浓度,计算血管通透指数(PPI);双抗夹心ELISA法对肺组织匀浆的细胞因子IFN-γ、IL-2、IL-4和IL-10水平进行检测,HE染色、Masson染色和羟脯氨酸含量研究肺部炎症及纤维化改变;采用免疫组化法测定肺组织IFN-γ及IL-4蛋白表达水平。结果与正常组比较,模型组大鼠白细胞数目和PPI均显著升高(P <0. 05),肺组织匀浆液细胞因子IFN-γ、IL-2、IL-4和IL-10释放增加,HE染色、Masson染色和羟脯氨酸含量结果显示大鼠造模成功;免疫组化结果显示模型大鼠IFN-γ、IL-4在肺组织表达上调。阳性药和复方银杏颗粒治疗后,大鼠白细胞数目和PPI均不同程度降低(P <0. 05),肺组织匀浆液细胞因子IFN-γ、IL-2、IL-4和IL-10释放减少,HE染色、Masson染色和羟脯氨酸含量结果显示大鼠纤维化停止甚至减轻;免疫组化结果显示6组大鼠IFN-γ、IL-4蛋白在肺组织表达下调。结论复方银杏颗粒明显改善肺间质纤维化,其机制可能与调控Th1/Th2信号通路有关。

Objective To observe the effects of compound recipe Gingko granules on rat models with pulmonary fibrosis,and to explore its possible action mechanism.Methods A total of 50 male Sprague-Dawley rats were injected intratracheally with bleomycin A5(5mg/kg)to establish rat pulmonary fibrosis models,then the model rats were randomly divided into model group,prednisone intervention group,high-dose,middle-dose,low-dose compound recipe Gingko granules groups,with 10 rats in each group,moreover,the other 10 rats were enrolled as control group.The rats in control group and model group were given normal by gavage,and the rats in prednisone intervention group were given prednisone 10ml/mg(0.32mg/ml),and the rats in compound recipe Gingko granules groups were given different doses of compound recipe Gingko granules(15g/kg,7.5g/kg,3.75g/kg,respectively)by gavage.After the rats were sacrificed the alveolar lavage fluid was collected and the leukocytes were counted.The BALF protein and serum total protein concentration were measured to calculate the vascular permeability index(PPI).The levels of cytokines IFN-γ,IL-2 and IL-4 The levels of IL-10 and IL-10 were detected by ELISA,and the changes of lung inflammation and lung fibrosis were detected by HE staining,Masson staining and hydroxyproline content detection.The expression levels of IFN-γand IL-4 in lung tissues were detected by immunohistochemistry.Results As compared with those in control group,the number of white blood cells and vascular permeability index in model group were significantly increased(P<0.05),and the levels of IFN-γ,IL-2,IL-4 and IL-10 in homogenate of lung tissues were increased.The HE staining,Masson staining and hydroxyproline content detection showed that the rat models were successfully established.Immunohistochemistry results showed that the expression levels of IFN-γand IL-4 in lung tissues were up-regulated in model rats.The number of white blood cells and vascular permeability index of rats in in prednisone intervention group and compound recipe Gingko granules groups were decreased to different extents(P<0.05).Moreover the levels of IFN-γ,IL-2,IL-4 and IL-10 in homogenate of lung tissues were increased.The results of HE staining,Masson staining and hydroxyproline content detection showed that the lung fibrosis of rats alleviated and even stopped.The results of immunohistochemistry showed that the expressions of IFN-γand IL-4 in lung tissue were down-regulated in all groups.Conclusion The compound recipe Gingko granules can significantly improve pulmonary interstitial fibrosis,and its action mechanism may be related to the regulation of Th1/Th2 signaling pathway.