右归饮对肺纤维化大鼠前列腺素及其受体的影响

Effect of Youguiyin on prostaglandin and its receptor in rats with pulmonary fibrosis

目的探讨前列腺素及其受体EP2/EP3在右归饮改善大鼠肺纤维化中的作用机制。方法 SPF级雄性SD大32只采用随机数字表完全随机法分为正常组、模型组、右归饮低剂量组(右低组)、右归饮高剂量组(右高组)各8只。正常组无任何干预,模型及用药组经气管插管雾化博来霉素(6 mg/kg)建立实验性肺纤维化大鼠模型。造模后第14天右低组和右高组分别定时给予5 g/kg、10 g/kg右归饮灌胃1次,模型组等量饮用水灌胃,连续给药20 d。记录大鼠肺脏质量和体重,计算肺脏系数,ELISA法检测血清和肺组织中前列腺素E2(PGE2)的浓度,免疫组织化学染色分析肺组织中EP2和EP3的表达情况以及组织定位,分别计数阳性细胞数目。结果与正常组相比,模型组大鼠肺脏质量(3. 1±0. 44 g,P <0. 05)和肺脏系数(0. 69±0. 13%,P <0. 05)均病理性升高;血清PGE2为45. 49±7. 38 pg/ml(P <0. 05),肺组织中PGE2为113. 64±16. 78 pg/ml(P <0. 05),均显著升高;与正常组相比,模型大鼠肺组织实变区内可见大量EP2强阳性的AECⅡs异常增生,肺泡管和肺泡腔内大量聚集的巨噬细胞部分呈EP3阳性,EP2阳性和EP3阳性的细胞数量均显著增加(P <0. 05);与模型组比较,右低组大鼠肺脏质量(2. 63±0. 24 g,P <0. 05)和肺脏系数下降(0. 57±0. 08%,P <0. 05),血清中的PGE2(23. 01±8. 61 pg/ml,P <0. 05)和肺组织的PGE2(89. 84±20. 54 pg/ml,P<0. 05)水平均降低,EP2阳性的Ⅱ型肺泡上皮细胞(AECⅡs)细胞EP2表达减弱,细胞数量减少(P <0. 05),而EP3阳性的巨噬细胞数增加(P <0. 05),右高组除血清中的PGE2水平降低(P <0. 05)外,其他未见统计学差异。结论右归饮能够明显改善博来霉素所致大鼠肺纤维化,可能与降低肺脏组织中PGE2水平,抑制实变区EP2的表达,减少EP2阳性的AECⅡs数量,增加EP3阳性的巨噬细胞有关。

Objective To investigate the role of the PGE2 and EP2/EP3 receptor on improving lung function of rats with pulmonary fibrosis treated with Youguiyin Decoction.Methods 32 SPF SD male rats were divided into normal group,model group,Youguiyin low dose group and Youguiyin high dose group,according to the random number table method.The normal group(8 rats)was given no intervention and was given normal feeding.The other 3 groups were established by intravenous injection of Bleomycin(6 mg/kg)via tracheal intubation.Youguiyin low dose group(8 rats)and Youguiyin high dose group(8 rats)were given 5 g/kg and 10 g/kg of Youguiyin Decoction at specific time from the 14th day after model establishment.The model group(8 rats)was given the same dose of drinking water once a day for 20 days.The lung weight and body weight were recorded,and the lung coefficient was calculated.The concentration of PGE2 in serum and lung tissue was measured with ELISA.The expression of EP2 and EP3 in lung tissue and tissue localization were analyzed by immunohistochemical staining,and the number of positive cells was counted.Results Compared with the normal group,the lung weight(3.1±0.44 g,P<0.05)and lung coefficient(0.69±0.13%,P<0.05)of the model group increased pathologically,and the level of PGE2 increased in both serum(45.49±7.38 pg/ml,P<0.05)and lung tissue(113.64±16.78 pg/ml,P<0.05).Abnormal proliferation of AEC IIs associated with strong EP2-positive in the consolidation of lung tissues,and the macrophages accumulated in the alveolar ducts and alveolar spaces were EP3-positive.The positive cells in the two groups increased significantly(P<0.05).Compared with the model group,the lung weight(2.63±0.24 g,P<0.05)and lung coefficient(0.57±0.08%,P<0.05)of Youguiyin low dose group decreased,and the level of PGE2 decreased in both serum(23.01±8.61 pg/ml,P<0.05)and lung tissue(89.84±20.54 pg/ml,P<0.05).The expression of EP2 in the AEC IIs with strong EP2-positive decreased,and the number of cell reduced(P<0.05).The number of macrophages with EP3-positive increased(P<0.05).In the Youguiyin high dose group,there was no statistical difference except the decreased level of PGE2 in serum(P<0.05).Conclusion Youguiyin can significantly improve pulmonary fibrosis induced by Bleomycin in rats,which may be associated with lowing PGE2 level in the consolidation of lung tissues,inhibiting the expression of EP2 in the consolidation area,reduction of AECIIs with EP2-positive,and increase of macrophages with EP3-positive.