平喘方对支气管哮喘模型小鼠巨噬细胞炎性蛋白-1α水平及CD86分子表达的实验研究

Experimental Effects of "Pingchuan Decoction" on Bronchial Asthma Macrophage Inflammatory Protein-1α and Expression of CD86 in Mice

目的通过研究哮喘模型小鼠巨噬细胞炎性蛋白-1α水平及CD86分子表达的影响,探索平喘方防治支气管哮喘的作用机制。方法通过对BALB/C小鼠用卵蛋白等腹腔注射致敏、雾化吸入激发,建立支气管哮喘小鼠模型。150只BALB/C小鼠随机分为5组,在激发哮喘7d、14d、21d时,用双抗体夹心ABC-ELISA法检测外周血和BALF中巨噬细胞炎性蛋白-1α(MIP-1α)水平,直接免疫荧光流式细胞术检测外周血和BALF中CD86+、CD3-CD86+细胞百分率,观察肺组织病理学改变。结果哮喘激发7d:各治疗组外周血和BALF中MIP-1α水平、CD86+细胞百分率均明显低于模型组(P<0.01);外周血CD3-CD86+细胞百分率明显均明显低于模型组(P<0.01);哮喘激发14d:各治疗组外周血和BALF中CD3-CD86+细胞百分率均明显低于模型组(P<0.01);平喘方常规剂量组、地塞米松组外周血和BALF中MIP-1α水平均明显低于模型组(P<0.01);哮喘激发21d:各治疗组外周血和BALF中MIP-1α水平、CD3-CD86+细胞百分率均明显低于模型组(P<0.01)。肺组织病理显示,各治疗组细支气管壁结构完好,管腔内少量炎性渗出物,管周少量炎细胞浸润。肺组织胶原染色图像分析显示,各治疗组面积与光密度乘积较模型组明显减少(P<0.01)。结论平喘方能显著降低MIP-1α水平、下调CD86分子表达、减轻气道黏膜上皮损伤,减少气道炎细胞浸润,减轻支气管壁及周围间质充血、水肿,具有减轻哮喘发病症状,改善哮喘气道炎症的作用。

Objective To study the effects of macrophage inflammatory protein-1α（MIP-1α） and expression of CD86 and explore the mechanism of ＂Pingchuan Decoction＂ on bronchial asthma. Methods To establish the model of asthmatic mice, the BALB/C mice were sensitized by intra-abdominal injection of mixed ovalbumin, and then activated by inhaling ovalbumin. One hundred and fifty BALB/C mice were randomly divided into control group, model group, normal-dosage ＂Pingchuan Decoction＂ group, qow-dosage ＂Pingchuan Decoction＂ group and Dexamethasone group. In the experiment, the level of MIP-1α in blood and BALF were measured by ABC-ELISA; direct immunfluorescence flow cytometry was used to detect CD86＋cell, CD3-CD86＋ cell percentage in blood and BALF; general histological changes of lung tissue were observed by hematoxylin and eosin stained sections on the 7th day, 14th day and 21st day. Results Compared with the control group, the levels of MIP-1α, the CD86＋cell, CD3-CD86＋ cell percentage in blood and BALF were raised significantly （P〈0.01）; CD86＋cell percentage in blood and BALF of any treatment group were significantly lower in the model group （P〈0.01）. The levels of MIP-1α, the CD86＋cell, the CD3-CD86＋ cell percentage in blood and BALF were of no difference between any two treatment groups （P〉0.05）. As compared with the control group, the levels of MIP-lot and CD3-CD86＋ cell percentage in blood and BALF were decreased significantly （P〈0.01）; the CD3- CD86＋ cell percentage in blood was significantly lower than the model group （P〈0.01） on the 7th day. The CD3-CD86＋ cell percentage in blood and BALF of every treatment group was lower than the model group （P〈0.01）. The levels of MIP-lot in blood and BALF of normal-dosage ＂Pingchuan Decoction＂ group and Dexamethasone group were lower than the model group （P〈0.01） on the 14th day. The levels of MIP-lot and the CD3-CD86＋ cell percentage in blood and BALF of every treatment group were lower than the model group （P〈 0.01） on the 21st day. In airway remodeling experiment, compared with the control group, the levels of MIP-1α, the CD86＋cell, CD3- CD86＋ cell percentage in blood and BALF and the levels of IgE in serum and BALF were raised significantly （P〈O.O1）. The level of MIP-lot, the CD86＂ cell, the CD3-CD86~ cell percentage in blood and BALF, the level of MIP-lot in serum and BALF were of no difference between any two treatment groups （.P〉0.05）. The collagen deposition was increased in model group, decreased in every treatment group compared with the model group. Conclusion The model mice sensitized by intra-abdominal injection of mixed ovalbumin and activated by inhaling ovalbumin had higher level of MIP-lot and the expression of CD86. ＂Pingchuan Decoction＂ can decrease the level of MIP-lot and the expression of CD86, improve chronic airway inflammation and inhibit the airway remodeling.