消乳散结胶囊对乳腺癌癌前病变大鼠乳腺组织CXCR4及VEGF表达的影响

The Influence of Capsule for Dispersing Breast Mass on the Expression of CXCR4 and VEGF in Breast Tissue of Rats with Precancerous Lesion of Breast Cancer

目的:观察消乳散结胶囊对乳腺癌癌前病变大鼠乳腺组织基质细胞衍生因子受体4(cxcchemokin receptor 4,CXCR4)、血管内皮生长因子(vascular endothelial growth factor,VEGF)基因及蛋白表达的干预作用。方法:将40只大鼠随机分为空白组、模型组、西药组和中药组,每组各10只。除空白组外,其余各组均用二甲基苯蒽(dimehylpheny lanthracene,DMBA)结合慢性不可预见应激行为制备乳腺癌癌前病变肝郁痰凝证模型。造模成功后,空白组及模型组大鼠给予蒸馏水10 mL·kg^-1灌胃;西药组大鼠给予枸橼酸他莫昔芬水溶液1.8 mg·kg^-1灌胃,中药组大鼠给予消乳散结胶囊水溶液0.324 g·kg^-1灌胃,连续灌胃30 d。HE染色观察乳腺组织病理变化,ELISA法检测血清孕酮(progesterone,P)、雌二醇(estradiol,E_2)变化情况,Real-time PCR及Western blot法检测各组大鼠VEGF、CXCR4 mRNA及蛋白表达水平。结果:西药组、模型组、中药组肿瘤发生率高于空白组,差异均有统计学意义(P<0.05);中药组肿瘤发生率高于模型组,差异具有统计学意义(P<0.05)。空白组、西药组和中药组E_2水平均低于模型组,差异有统计学意义(P<0.05)。各组P水平比较,差异无统计学意义(P>0.05)。模型组、西药组VEGF mRNA表达及蛋白水平低于空白组,差异有统计学意义(P<0.01),中药组与空白组比较,差异无统计学意义(P>0.05)。西药组、中药组VEGF、CXCR4 mRNA表达及蛋白水平均低于模型组,差异均有统计学意义(P<0.05)。结论:消乳散结胶囊可能是通过调整E_2分泌,并抑制VEGF及CXCR4基因表达,从而抑制新生血管生成实现控制大鼠乳腺癌癌前病变的作用。

Objective:To observe the intervention effect of Capsule for Dispersing Breast Mass on the gene of cxcchemokin receptor 4(CXCR4)and vascular endothelial growth factor(VEGF)and protein expression in breast tissue of precancerous lesion rats.Methods:A total of forty rats were randomly divided into the blank group,the model group,the western medicine group and the TCM group,with 10 rats in each group.Except the blank group,DMBA combined with chronic unpredictable stress behavior was used to establish the model of liver depression and phlegm coagulation in precancerous lesions of breast cancer.After successful modeling,the rats in the blank group and the model group were given distilled water 10 mL·kg-1 by gavage;the rats in the western medicine group were given tamoxifen citrate aqueous solution 1.8 mg·kg-1 by gavage;the rats in the TCM group were given aqueous solution of Capsule for Dispersing Breast Mass by gavage.The treatment course lasted for 30 days.HE staining was used to observe the pathological changes of breast tissue.ELISA was used to detect the changes of serum progesterone(P)and estradiol(E2).Real time PCR and Western blot were used to detect the protein expression levels of VEGF and CXCR4 mRNA.Results:The incidence of tumor in the western medicine group,the model group and the TCM group was higher than that in the normal group,and all the differences were statistically significant(P<0.05).The incidence of tumor in the TCM group was higher than that in model group,and the difference was statistically significant(P<0.05).The E2 levels in the blank group,the western medicine group and the TCM group were lower than those in the model group,and all the differences were statistically significant(P<0.05).There was no significant difference in P level among the groups(P>0.05).The expression of VEGF mRNA and protein level in the model group and the western medicine group were lower than those in the blank group,and all the differences were statistically significant(P<0.01),but there was no significant difference between the TCM group and the blank group(P>0.05).VEGF and CXCR4 mRNA expression and protein levels of the western medicine group and TCM group were lower than those in the model group,and all the differences were statistically significant(P<0.05).Conclusion:Capsule for Dispersing Breast Mass may control precancerous lesions of breast cancer in rats by regulating E2 secretion and inhibiting the expression of VEGF and CXCR4 genes,thus inhibiting angiogenesis.