低分子肝素对大鼠COPD模型肺组织中肝细胞生长因子的影响

Effect of low molecular weight heparin on hepatocyte growth factor in lung tissue of rats with COPD

目的探讨低分子肝素(LMWH)对大鼠慢性阻塞性肺疾病(COPD)模型肺组织中肝细胞生长因子(HGF)的影响。方法选清洁级雄性Wistar大鼠30只,通过随机数字表法将大鼠分为对照组、COPD组、LMWH组各10只。对照组大鼠于正常环境下喂养,COPD组与LMWH组大鼠通过熏烟及注入脂多糖建立COPD模型,LMWH组熏烟前0.5 h皮下注射LMWH 1 ml,熏烟结束后取右肺上叶组织于组织液氮中保存,通过免疫印迹法检测肺组织VEGF、HGF、c-met蛋白水平,通过酶联免疫吸附法检测肺组织转化生长因子β(TGF-β)蛋白水平。通过逆转录-聚合酶链式反应法检测肺组织血管内皮生长因子(VEGF)、HGF、肝细胞生长因子受体(c-met)mRNA水平,对比3组大鼠VEGF、HGF、c-met蛋白及VEGF、HGF、c-met mRNA及TGF-β水平。结果COPD组合LMWH组肺组织中VEGF、HGF、c-met蛋白水平和VEGFmRNA、HGFmRNA、c-metmRNA水平以及TGF-β水平均明显高于对照组(P值均<0.05),而LMWH组肺组织中以上指标均明显低于COPD组(P值均<0.05)。结论COPD大鼠VEGF、HGF、c-met、TGF-β水平呈现出高表达,上述因子可能参与COPD的发生、发展,而LMWH则能对其水平表达有抑制作用。

Objective To investigate the effect of low molecular weight heparin(LMWH)on hepatocyte growth factor(HGF)in lung tissue of rats with chronic obstructive pulmonary disease(COPD).Methods A total of 30 clean grade male Wistar rats were randomly divided into control group,COPD group and LMWH group with 10 rats in each group.The rats in the control group were fed in the normal environment.COPD models were established by smoking and injecting lipopolysaccharide in COPD group and LMWH group.LMWH group was subcutaneously injected with LMWH 0.5 h before smoking.After smoking,the right upper lobe of lung tissue was taken and stored in tissue liquid nitrogen.The protein levels of VEGF,HGF and c-met in lung tissue were detected by Western blot,protein levels of transforming growth factorβ(TGF-β)in lung tissues were determined by enzyme-linked immunosorbent assay.The mRNA levels of vascular endothelial growth factor(VEGF),HGF and hepatocyte growth factor receptor(c-met)in lung tissue were detected by reverse transcription polymerase chain reaction(RT-PCR),VEGF,HGF,c-met protein,VEGF,HGF,c-met mRNA and TGF-βlevels were compared among three groups.Results The levels of VEGF,HGF,c-met protein and the levels of VEGFmRNA,HGFmRNA,c-metmRNA and TGF-βin the lung tissues of the COPD and LMWH groups were significantly higher than those of the control group(all P<0.05),while the above indicators in the lung tissues of the LMWH group All were significantly lower than the COPD group(all P<0.05).Conclusions The levels of VEGF,HGF,C-MET,and TGF-βin COPD rats are highly expressed,and these factors may participate in the development of COPD,while LMWH can influence the expression level.