黄芪和当归各提取物优化配方对特发性肺纤维化小鼠生存状况及肺细胞新生相关基因表达水平的影响

【目的】观察黄芪当归各提取物优化配方对特发性肺纤维化(IPF)小鼠生存状况、转化生长因子(TGF-β)、血管内皮生长因子(VEGF)表达水平的影响,确定最佳治疗方案,并探讨其内在机理。【方法】选用SPF级ICR雄性小鼠105只,随机分为正常组、模型组、5个中药治疗组(即黄芪当归组分优化方1~5组)。模型组和治疗组于气管内注射博莱霉素(剂量为5 mg/kg)复制小鼠肺纤维化模型,在第21天时剪取小鼠肺组织,采用碱水解法检测羟脯氨酸(HYP)的含量;苏木素—伊红(HE)染色及Mallory氏染色,观察肺组织形态变化;聚合酶链反应(PCR)技术检测肺细胞新生相关基因转化生长因子(TGF)-β、血管内皮生长因子(VEGF)表达水平。【结果】HE染色及Mallory氏染色结果显示,5个中药治疗组较模型组肺纤维化程度均有所改善,其中优化方1组改善明显,肺泡结构恢复较好。治疗组小鼠的21 d死亡率较模型组均有所降低,且优化方1组、5组作用显著,差异有统计学意义(P0.05)。与模型组比较,治疗组小鼠肺组织HYP含量各有不同程度的降低,差异有统计学意义(P0.05或P0.01)。TGF-β、VEGF基因表达水平模型组均高于正常组,治疗各组(除优化方5组TGF-β表达外)较模型组有不同程度的降低,差异有统计学意义(P0.05或P0.01)。【结论】黄芪水提物和当归醇提物所占比例较大时,可降低肺中HYP的含量,抑制TGF-β、VEGF表达水平,从而改善小鼠肺纤维化的程度。

Effects of Optimized Formulas of Radix Astragali and Radix Angelicae Sinensis Extracts on Survival Status of Idiopathic Pulmonary Fibrosis Mice and on Expression of Cytogenesis-related Factors in Lung Tissues

Objective To observe the effect of the optimized formulas of extracts of Radix Astragali and Radix Angelicae Sinensis on the survival status of the idiopathic pulmonary fibrosis (IPF) mice,and on the expression levels of transforming growth factor-β (TGF-β) and vascular endothelial growth factor(VEGF),so as to optimize the therapeutic regimen and to explore the therapeutic mechanism.Methods One hundred and five SPF ICR male mice were randomly divided into normal group,model group and 5 Chinese medicine treatment groups (group 1,2,3,4,5 of the optimized formula of Radix Astragali and Radix Angelicae Sinensis extracts).The mice in the model group and the 5 treatment groups were intratracheally injected with bleomycin (5 mg/kg) to induce the pulmonary fibrosis model.On day 21,the lung tissues were taken out for the test.Hydroxyproline content was detected by alkaline hydrolysis method,and morphological changes of lung tissues were observed by hematoxylineosin (HE) staining and Mallory's staining methods.The expression levels of TGF-β and VEGF were detected by polymerase chain reaction (PCR).Results The HE staining and Mallory's staining results showed that the pulmonary fibrosis in the 5 treatment groups was relieved as compared with that in the model group,especially in the group 1,and the alveolar structure recovered better.The 21-day overall death rate in the treatment groups were lower than those in the model group,and group 1 and group 5 had the lowest rates,the difference being statistically significant (P＜ 0.05).Compared with the model group,the content of hydroxyproline in the lung tissues of the treatment groups were decreased to some degrees,and there was significant difference (P ＜ 0.05 or P ＜ 0.01).The expression levels of TGF-β and VEGF in model group were higher than those in normal group,but were deceased in the treatment groups to some degrees,except TGF-β expression in group 5,and the difference was significant(P ＜ 0.05 or P ＜ 0.01).Conclusion When the contents of Radix Astragali water-extract and Radix Angelicae Sinensis alcohol-extract were predominated,the extract formula exerts certain effects on decreasing hydroxyproline content in the lung tissues,inhibiting the expression levels of TGF-β and VEGF,and relieving the degree of pulmonary fibrosis in IPF mice.