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七拗汤对内毒素诱导加重型哮喘小鼠的作用和机理研究

The effect of Qi'ao Decoction on ovalbumin induced and lipopolysaccharide enhanced severe asthma mice and its mechanism

  • 摘要: 目的:评价七拗汤对哮喘小鼠气道炎症和高反应性的影响。方法:120只Balb/C小鼠随机分为6组:空白组、模型组、地塞米松组、七拗汤高剂量组、七拗汤中剂量组、七拗汤低剂量组。除空白组外,各组小鼠以皮下注射、腹腔注射OVA致敏,雾化吸入OVA激发,内毒素鼻腔滴入诱导。空白组小鼠用PBS进行致敏、激发并在气道内滴入PBS。七拗汤高、中、低剂量组分别予26.8、13.4、6.7 gkg-1的七拗汤灌胃;地塞米松组予0.002 4 gkg-1的醋酸地塞米松溶液,1次/天,连续7天;末次滴鼻激发24 h后测定相关指标。结果:模型组中嗜酸性粒细胞和中性细胞数显著上升(P0.01);与模型组相比,地塞米松组嗜酸性粒细胞、中性细胞数量降低(P0.05,P0.01)。七拗汤高、中、低剂量组嗜酸性粒细胞水平均有下降(P0.01)。模型组气道反应性升高,与模型组相比,地塞米松组、七拗汤高、中、低剂量组均有所降低(P0.01)。病理显示,模型组小鼠肺组织炎细胞浸润,主要是中性粒细胞和嗜酸性粒细胞,并见气管壁增厚。与模型组相比,地塞米松组病变明显减轻,仅见轻度的支气管周围炎;七拗汤高、中、低剂量组肺泡壁、气道壁病变程度均较模型组减轻,管壁及其周围浸润的炎细胞数量减少,支气管上皮细胞变性不明显,支气管腔内渗出物不同程度地减少。模型组IFN-和IL-12水平显著降低,IL-4上升(P0.05);与模型组相比,地塞米松组IFN-和IL-12水平升高,IL-4降低(P0.05)。七拗汤高、中剂量组中IFN-水平升高,中剂量组中IL-4和IL-12水平与模型组相比具有统计学意义(P0.05);七拗汤低剂量组细胞因子水平改变与模型组相比无意义。结论:七拗汤能够显著抑制卵蛋白致敏激发结合内毒素诱导的加重型哮喘小鼠的气道炎症和气道高反应性,通过调整其细胞因子水平,改善肺组织病理结构等发挥治疗效果。

     

    Abstract: AIM:To evaluate the effect of Qi'ao Deocoction(QAD) on the inflammation and hyperresponsiveness of asthma mice.METHODS:120 Balb/C mice were randomly divided into six groups:normal group,model group,dexamethasone group,high dose QAD group,medium dose QAD group and low dose QAD group.The asthma model was reproduced in Balb/C mice sensitized by ovalbumin,challenged by OVA and LPS.The mice of the normal group were sensitized,chanllenged and intranasally instilled by PBS.On day 28-34,6.7,13.4 and 26.8 gkg-1 Qi'ao Decoction were administrated;0.002 4 gkg-1 dexamethasone solution was given to the dexamethasone group;normal and model groups were given the same amount of normal saline.Bronchoalveolar lavage fluid,airway hyperresponsiveness,lung histopathology and cytokines were then collected and analyzed.RESULTS:Compared with normal group,total cellular score,the number of macrophages,lymphocytes,eosinophils and neutrophils of model group significantly increased(P0.01).Compared with model group,the administration of dexamethasone induced a significant decrease in eosinophils and neutrophils(P0.05,P0.01).The number of eosinophils,which plays an important role in airway inflammatory reaction of asthma,of the three QAD groups all decreased(P0.01).RL before and after Ach(5 mgmL-1) stimulation in the model group both overtook that in the normal group(P0.01).Compared with model group,dexamethasone group,high dose QAD group,medium dose QAD group and low dose QAD group groups all had significantly lower RL before and after Ach stimulation(P0.01).Normal pulmonary histopathology was found in the normal group.In the model group,mice exhibited marked increases in inflammatory cell infiltration,mostly including neutrophils and macrophages,perivascular inflammation and thickened alveolus wall(P0.01).Dexamethasone application mitigated inflammation around the bronchi(P0.05).These histopathological changes were ameliorated in the three decoction groups(P0.01,P0.05).In addition,alveolus and airway wall lesions of medium dose QAD group and high dose QAD group were reduced,the number of inflammatory cells infiltrated around the walls decreased,no clear degeneration of bronchial epithelial cells was found,and exudates in bronchi declined in different degrees.Compared with normal group,IFN- and IL-12 of model group significantly decreased,while IL-4 increased,showing statistic difference(P0.05).Compared with model group,IFN- and IL-12 level of dexamethasone group went up too,but IL-4 declined(P0.05).The level of IFN- of medium dose QAD group and high dose QAD group both increased;IL-4 and IL-12 of medium dose group were found significant differences(P0.05);but none of the cytokines of low dose QAD group showed statistical significance(P0.05).CONCLUSION:QAD can significantly inhibit airway inflammation and airway hyperresponsiveness of mice with severe asthma induced by ovalumin and lipopolysaccharide,adjust the balance of cytokines,and improve lung histopathological condition.So,it exhibits great effect on severe asthma.

     

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