黄芪当归对药对特发性肺纤维化小鼠生存状况及组织修复相关基因表达水平的影响

Influence of the pair drugs of astragalus and angelica on the IPF living conditions and tissue repair related gene expression level in mice

目的观察不同剂量与配伍比例的黄芪、当归对药对特发性肺纤维化(idiopathic pulmonary fibrosis,IPF)小鼠生存率、体质量及组织修复相关基因c-kit原癌基因及碱性成纤维细胞生长因子(fibroblast growth factor2,FGF2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达水平的影响,并探讨其相互关系及保护机制。方法 SPF级ICR雄性小鼠80只,随机分为正常对照组、模型组,以及6个中药治疗组(大剂量黄芪50 g当归10g组、大剂量黄芪30 g当归30g组、大剂量黄芪10 g当归50 g组、小剂量黄芪25 g当归5 g组、小剂量黄芪15 g当归15 g组、小剂量黄芪5 g当归25 g组),除正常对照组外,其余各组采用气管内注射博莱霉素(5 mg/kg)复制小鼠肺纤维化模型,于28天处死小鼠,采用real time-PCR法检测组织修复相关基因c-kit、FGF2及VEGF表达水平,HE染色及武兆发简化Mallory氏染色,观察肺组织形态变化。结果 HE染色及武兆发简化Mallory氏染色显示,6个中药治疗组较模型组均有所改善,其中中药治疗第1、6组改善明显,肺泡结构已基本恢复。第28天时,与模型组比较,各组小鼠体质量均呈上升的趋势,中药第1组小鼠体质量上升趋势较大(P<0.05)。28天死亡率,中药治疗组小鼠的死亡率较模型组降低,其中第1组死亡率与模型组的差异有统计学意义(P<0.05)。模型组c-kit、FGF2、VEGFm RNA表达水平均高于对照组(P<0.05)。结论从小鼠的生存率与体质量变化及病理形态观察,大剂量黄芪当归5∶1组能明显改善小鼠的生存质量,其作用机制可能与抑制VEGF m RNA的表达水平,促进c-kit及FGF2 m RNA的表达水平有关。

Objective To observe the influence of different doses and compatibility proportion of the pair drugs of astragalus and angelica on the survival rate,weight and tissue repair kit and FGF2 c- related genes,level of VEGF expression of IPF mice and discuss the relationship between the pair drugs and the protection mechanism. Methods 80 SPF ICR male mice were randomly divided into the normal control group,the model group,the traditional Chinese medicine treatment group and 6（ Three groups of large dose of astragalus membranaceus and angelica： 1. 50 g and 10 g; 2. 30 g and 30 g; 3. 10 g and 50 g.Three groups of small doses of Astragalus and Angelica： 4. 25 g and 5 g; 5. 15 g and 15 g; 6. 5 g and 25g）. Except the normal control group,other groups by intratracheal injection of bleomycin（ 5mg / kg） to replicate the model of pulmonary fibrosis in mice,the mice were sacrificed after 28 days. The expression levels of the repair of FGF2 and VEGF related gene c-kit were tested by RT-PCR; the morphological changes oflung tissue were observed by HE staining and Wu Zhaofa simplified Mallory＇s staining. Results The result of HE dyeing and reduction Wu Zhaofa Mallory showed that six Chinese medicine treatment groups were improved compared to the model group and the Chinese medicine group of 1 and 6 showed significant improvement,and the alveolar structure have been restored. Compared to the model group,mice weight of each group showed ascendant trend,and Chinese medicine group of 1 showed significantly larger trend of weight gain in mice（P〈0. 05）. Mice mortality was reduced in the Chinese medicine treatment groups compared to the model group,and the group 1 showed significant difference（P〈0. 05）. Expression levels of c-kit,FGF2,VEGF m RNA in the model group were significantly higher than the control group（P〈0. 05）. Conclusion The results showed that the large dose of astragalus and angelica group of 5∶ 1 can obviously improve the quality of the survival of mice and its mechanism may be related to inhibit the expression of VEGF m RNA level,and promote the c-kit and FGF2 m RNA expression level.