[关键词]
[摘要]
目的研究中成药润肺止嗽丸对慢性阻塞性肺疾病(COPD)大鼠细胞凋亡的影响,并探讨其作用机制。方法 32只雄性Wistar大鼠随机分为对照组、模型组、润肺1月和3月组;使用烟熏联合气道滴入脂多糖法制备COPD大鼠模型;润肺1、3月组ig给予润肺止嗽丸0.9 g/kg,每天给药1次,分别连续给药1、3个月,对照组和模型组给予等体积的超纯水1个月;HE染色观察慢阻肺大鼠肺组织变化;TUNEL法检测凋亡细胞并计算凋亡指数(AI);Elisa法检测肺泡灌洗液(BALF)中血管内皮生长因子(VEGF)水平;Western blotting法检测肺组织中VEGF和血管内皮生长因子受体2(VEGFR2)的蛋白表达。结果与对照组比较,模型组大鼠肺组织形态病变明显;AI显著升高;BALF中VEGF、肺组织中VEGF和VEGFR2水平均显著下降。与模型组比较,润肺1月和3月组肺组织形态明显好转;AI显著下降;BALF中VEGF、肺组织中VEGF和VEGFR2水平显著升高;且润肺3月组效果好于1月组。结论润肺止嗽丸可以改善慢阻肺细胞凋亡,机制可能与上调VEGF和VEGFR2相关。
[Key word]
[Abstract]
Objective To study the effect of Runfei Zhisou Pills (RZP) on cell apoptosis in rat model of chronic obstructive pulmonary disease (COPD) and its mechanism. Methods Totally 32 male Wistar rats were randomly divided into control group, model group, 1-month RZP group, and 3-month RZP group. Rats in 1-month and 3-month RZP group, received RZP of 0.9 g/kg once daily for 1 and 3 months respectively, and rats in control and model groups received water with equal volume for 1 month. The rat COPD model was established using tobacco smoke combined with intratracheal instillation of LPS. Morphological changes of lung tissue in COPD rats were observed with HE staining under light microscope. The VEGF levels in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The VEGF and VEGFR2 protein expression in lung tissues was measured by Western blotting. Cell apoptosis in lung tissues was detected with TUNEL method, and apoptosis index (AI) was counted. Results Compared with control group, the pathological changes of lung tissue in model group were obvious, the AI significantly increased, VEGF in BALF and VEGF and VEGFR2 in lung tissue decreased significantly. Compared with model group, morphological improvement of lung tissue was obvious, the AI significantly decreased, and VEGF in BALF, VEGF and VEGFR2 in lung tissue increased significantly in 1-month and 3-month RZP groups. Conclusion RZP can inhibit cell apoptosis of COPD, and the mechanism may be related with up-regulation of VEGF and VEGFR2.
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[基金项目]
河北省卫生厅基础研究计划专项项目(2014131015)