Effects of Taohong Siwu Decoction on the expression of pathological angiogenesis correlation factors including VEGF,KDR and Flt-1 in hepatic fibrosis mice
目的:从整体水平研究复方桃红四物汤对四氯化碳所致肝纤维化模型小鼠肝组织血管内皮生长因子(VEGF)、含激酶插入区受体(KDR)及Flt-1蛋白与mRNA表达的影响。方法:昆明小鼠60只,随机分组为空白组,模型组,秋水仙碱组,桃红四物汤高、中、低(17.00、8.50、4.25g/kg)剂量组,采用左侧腹股沟处皮下注射四氯化碳花生油溶液建立小鼠肝纤维化模型,给药6周后,运用酶联免疫吸附测定(ELISA)法检测肝组织VEGF蛋白含量;蛋白印迹法(Western blot)测定小鼠肝脏KDR、Flt-1蛋白的表达;逆转录PCR(RT-PCR)法测定小鼠肝脏VEGF、KDR及Flt-1 mRNA表达。结果:桃红四物汤治疗后,桃红四物汤高、中剂量组肝组织VEGF蛋白表达量较模型组降低(P<0.05,P<0.01),中药高剂量组肝组织KDR与Flt-1蛋白表达较模型组显著降低(P<0.05);而桃红四物汤高、中剂量组肝组织VEGF、KDR与Flt-1 mRNA表达较模型组、秋水仙碱组明显减少(P<0.01),并且呈现一定的量效正相关。结论:桃红四物汤能够降低肝纤维化病理血管生成相关因子的表达,减缓病理性的微血管生成可能是其改善肝纤维化的机制之一。Objective:To investigate the effects of Taohong Siwu Decoction(THSWD) on protein and mRNA expression of vascular endothelial growth factor(VEGF),kinase insert domain containing receptor(KDR) and Flt-1 in hepatic fibrosis model mice caused by CCI_4.Methods:Sixty Kunming mice were randomly divided into blank,model,colchicine(control)and three THSWD(17.00,8.50,4.25g/kg) groups,with 10 mice in each group.Except the blank group,the other groups' mice were subcutaneously injected with carbon tetrachloride(CCI_4)-arachis oil solution in left inguen to establish hepatic fibrosis mouse model.After administration for six weeks,the content of VEGF protein in liver tissue was measured by enzyme-linked immunosorbent assay(ELISA),protein expressions of KDR and Flt-1 in liver tissue were detected by Western blotting,and mRNA expressions of VEGF,KDR and Flt-1 in liver tissue were measured by reverse transcription polymerase chain reaction(RT-PCR).Results:After treatment,the content of VEGF protein in THSWD(17.00,8.50g/kg) groups were lower than that in model group(P<0.05,P<0.01).The protein expressions of liver tissue KDR and Flt-1 in THSWD(17.00g/kg) group were less than that in model group(P<0.05).The mRNA expressions of liver tissue VEGF,KDR and Flt-1 were significantly lower than that in both model and colchicine groups(P<0.01),which showed a certain positive correlation between dose and effect.Conclusion:Taohong Siwu Decoction could decrease the expression of pathological angiogenesis correlation factors of hepatic fibrosis,thus helpful to induce the formation of pathological capillary vessels,which may be one of the possible mechanisms of ameliorating hepatic fibrosis.国家自然科学青年基金项目(No.81202659);; 福建省自然科学基金项目(No.2014J01373);; 厦门大学新聘教师科研启动基金项目(No.ZK1014);; 福建省大学生创新创业训练项目(No.DC2013174)~
