泻肺定喘汤对中性粒细胞型哮喘大鼠KLF-2/RelA比例平衡及中性粒细胞凋亡的影响

Effect of Xiefei Dingchuan Decoction on KLF-2/RelA Proportion Balance and Neutrophil Apoptosis in Rats with Neutrophil Asthma

目的观察泻肺定喘汤对中性粒细胞型哮喘大鼠KLF-2/RelA比例平衡及中性粒细胞凋亡的影响。方法 48只大鼠随机分为空白组、模型组、地塞米松组和泻肺定喘汤低、中、高剂量组。除空白组外,其余组均按照脂多糖(LPS)+卵白蛋白(OVA)联合致敏的方法复制中性粒细胞型哮喘模型。造模后各药物干预组给予相应的药物治疗,空白组和模型组给予等体积的0.9%氯化钠注射液灌胃治疗,每日1次,连续2周。疗程结束后观察各组大鼠肺泡灌洗液中白细胞计数及分类,流式细胞术检测外周血中性粒细胞的凋亡,并通过Western blotting法测定中性粒细胞中KLF-2、RelA、Bax和Bcl-2蛋白的表达。结果与模型组比较,泻肺定喘汤中、高剂量均能够显著降低肺泡灌洗液中白细胞数量及中性粒细胞、嗜酸性粒细胞和淋巴细胞的百分比,增加中性粒细胞的凋亡,差异均具有显著统计学意义(P <0.01);此外,泻肺定喘汤中、高剂量还能显著降低中性粒细胞中Bcl-2蛋白的表达,上调KLF-2、RelA、Bax蛋白的表达和KLF-2/RelA、Bax/Bcl-2的比值,与模型组比较,差异均具有统计学意义(P <0.05或P <0.01)。结论泻肺定喘汤能够通过调控KLF2/RelA比例的平衡,从而促进中性粒细胞的凋亡,减轻炎症反应。

Objective: To observe the effect of Xiefei Dingchuan Decoction on the balance of KLF-2/RelA ratio and apoptosis of neutrophil cells in rats with neutrophil asthma. Methods: 48 rats were randomly divided into the control group,the model group,DXN group and XFD low,medium and high dose groups. Rats were sensitized by LPS+OVA to establish the neutrophil asthma model except the control group. After modeling,intervention groups were given the corresponding drug treatment,while the control group and the model group were treated with saline gavage,once a day for two weeks. After the treatment,the white blood cell count and classification in the alveolar lavage fluid were observed. The apoptosis of peripheral blood neutrophils was detected by flow cytometry. The expression of KLF-2,RelA,Bax and Bcl-2 protein in neutrophils was detected by Western blot. Results: Compared with the model group,XFD medium and high dose groups could significantly reduce the number of white blood cells,the percentage of neutrophils,eosinophils and lymphocytes in the bronchoalveolar lavage fluid,and increase the apoptosis of neutrophils,differences were statistically significant(P < 0.01). XFD medium and high dose groups could also significantly decrease the expression of Bcl-2 protein in neutrophils,up-regulated the expressions of KLF-2,RelA,Bax proteins and ratios of KLF-2/RelA and Bax/Bcl-2,with significant difference compared with the model group(P < 0.05 or P < 0.01). Conclusion: XFD can promote the apoptosis of neutrophils and alleviate inflammation by regulating the balance of KLF2 / RelA ratio.