止咳平喘合剂影响哮喘大鼠嗜酸粒细胞及Eotaxin表达的实验研究

Study of Effects of Zhike Pingchuan Mixture on EOS and Eotaxin Expression in Asthmatic rats

目的:观察中药止咳平喘合剂对哮喘大鼠肺内嗜酸粒细胞(Eosinophil,EOS)及嗜酸粒细胞趋化因子(Eotaxin)表达水平的影响,探讨中药止咳平喘合剂治疗哮喘的可能作用机制。方法:用改良的卵白蛋白多点注射致敏与雾化吸入激发法建立大鼠哮喘模型。将60只雄性SD大鼠随机分为6组,每组均10只,分别为正常对照组、哮喘模型组、强的松组、止咳平喘合剂小剂量组、中剂量组和大剂量组。实验第15天起灌胃给药,根据等效剂量换算比率计算,正常对照组、哮喘模型组分别给予生理盐水10mL/kg,强的松组给予强的松混悬液10mg/kg,止咳平喘合剂小剂量组7.5g/kg,中剂量组15g/kg,大剂量组30g/kg,给药连续5天,每天1次。末次雾化激发后24h处死大鼠留取标本。镜下观察各组哮喘大鼠肺组织病理形态的变化及支气管肺泡灌洗液(BALF)中白细胞、EOS计数的变化;免疫组化染色法检测肺组织Eotaxin蛋白的表达水平。结果:哮喘模型组大鼠BALF中EOS数量明显增高,与其它各组相比,均存在显著性差异(P<0.01);止咳平喘合剂组大鼠肺组织气道黏膜及黏膜下层的炎性细胞浸润明显减轻,与哮喘模型组相比均有显著性差异(P<0.01);哮喘模型组大鼠肺组织Eotaxin蛋白表达量明显增高,与正常对照组及药物治疗组相比有显著性差异(P<0.01),止咳平喘合剂各剂量组之间无显著性差异(P>0.05)。结论:止咳平喘合剂可明显减轻哮喘大鼠气道炎症,其机制可能与其减低哮喘大鼠肺组织Eotaxin蛋白的表达相关。

Objectives： To observe the effects of Zhikepingchuan Mixture on Eosinophi｜ （EOS） and Eotaxin expression of allergic - induced asthmatic rat and investigate the mechanism if Zhikepingchuan Mixture treating asthma. Methods ： Asthma rats models were induced by modified intraperitioneal sensitization and challenged with nebulized ovalbumin. Sixty male SD rats were randomly divided into six experimental groups（ 10 in each）： normal control group, asthma model group, positive control group （prednisone group）, lowdose group of Zhikepingchuan Mixture, middle- dose group of Zhikepingchuan Mixture, high -dose group of Zhikepingchuan Mixture. From thelSth day ,The normal control group and the asthma model group received normal saline in 10mL/kg; the positive control group received prednisone in 10mg/kg; the low dose group received 7.5g/kg of Zhikepingchuan mixture （containing crude drug 2.0g/mL）, and middle dose group received 15g/kg, and high dose group 30g/kg. All drugs were given for 5 days, once a day. After excitation in the last 24h,rats in each group were all put to death. We intivigated the change of eosinophils count, the total count of leuko- cyte in BALF and the change of lung tissue of each group by microscope. The expression of eotaxin in the lung tissues was detected by immunohistnchemical staining. Results： The count of EOS and the total count of leukocyte in BALF of the asthma model group were significantly increased compared with those of normal group and each treatment group （P 〈 0.01 ）. The eotaxin expression of asthma model group in the lung tissue significantly increased compared with each treat- ment group（P 〈 0.01 ）. The three dose groups of Zhikepingchuan Mixture had no statistical difference（ P 〉 0.05）. Conclusions ： Zhikepingchuan Mixture also can decrease the expression of eotaxin in the lung tissue. So Zhikepingchuan Mixture has the role of restraining airway inflammatory in asthmatic rats.