花姜酮对激素不敏感哮喘小鼠的治疗作用及其机制

Effect and underlying mechanism of zerumbone on mice with hormone insensitive asthma

目的探讨花姜酮对激素不敏感哮喘小鼠的治疗作用及其可能机制。方法将30只BALB/c小鼠随机均分为五组:第0、1、2、7天,B、C、D、E组用75μg鸡卵清蛋白+10μg脂多糖滴鼻致敏,A组用PBS滴鼻致敏;第14、15、21、22天,五组小鼠均用50μg鸡卵清蛋白滴鼻激发;每次激发30min前,A、B、C、D、E组小鼠分别腹腔注射200μl PBS、200μl PBS、花姜酮10mg/kg、花姜酮50mg/kg、地塞米松1mg/kg。采用瑞氏-吉姆萨复合染色进行支气管肺泡灌洗液(BALF)中细胞分类计数,HE染色观察气道炎症的改变,ELISA法检测BALF及血清中的IL-17水平,流式细胞术检测小鼠脾脏中Th17细胞的比例。结果与A组相比,B组小鼠气道炎症加重,IL-17水平、Th17细胞的比例以及总细胞、嗜中性粒细胞和嗜酸性粒细胞计数均增加(P<0.05),而C组和D组能抑制上述各指标的增加过程(P<0.05),其中D组的抑制作用更为明显(P<0.05)。结论花姜酮能有效治疗激素不敏感哮喘小鼠;其机制可能是通过抑制Th17细胞及IL-17介导的中性粒细胞性气道炎症。

Objective To investigate the therapeutic effect and underlying mechanism of zerumbone on the mice with hormone insensitive asthma.Methods Thirty BALB/c mice were equally randomized into five groups.The mice were sensitized with 75μg ovalbumin and 10μg lipopolysaccharide by nasal drip in groups of B,C,D and E,which were sensitized with PBS in group A on the 0,1 st,2 nd and 7 th day.Then the mice in all groups were challenged with 50μg ovalbumin by nasal drip on the 14 th,15 th,21 st and 22 nd day.Thirty minutes before challenging every time,the groups of A,B,C,D and E were intraperitoneally injected with 200μl PBS,200μl PBS,zerumbone 10 mg/kg,zerumbone 50 mg/kg and dexamethasone 1 mg/kg,respectively.The number of cell classification in bronchoalveolar lavage fluid(BALF)was determined by Wright-Giemsa staining.The change of airway inflammation was observed by HE staining.The levels of IL-17 in serum and BALF were detected by ELISA method.The proportion of Th17 cells in the spleen was measured by flow cytometry.Results Compared with group A,the airway inflammation of mice was worse,and the level of IL-17,the proportion of Th17 cells and the number of total cells,neutrophilic granulocyte and eosinophilic granulocyte were all significantly increased in group B(P<0.05),which were obviously inhibited by zerumbone in groups of C and D(P<0.05),especially in group D(P<0.05).Conclusion Zerumbone has an effective therapeutic effect on hormone insensitive asthma in mice probably via inhibiting Th17 cells and IL-17-mediated neutrophilic airway inflammation.