扶正抑瘤方联合替莫唑胺抑制大鼠脑胶质瘤生长及调控血肿瘤屏障相关蛋白的作用研究

摘要

目的 探讨扶正抑瘤方联合替莫唑胺对C6脑胶质瘤大鼠肿瘤生长及血肿瘤屏障相关蛋白(zonula odluden,ZO-1)、P-糖蛋白(glycoprotein,gp)及细胞质膜微囊蛋白Caveolin-1的影响.方法 雄性Wistar大鼠50只,随机分为5组:假手术组、模型组、替莫唑胺组、扶正抑瘤方组和扶正抑瘤方联合替莫唑胺组.采用立体定位法建立C6大鼠脑胶质瘤模型,造模当天开始,各组大鼠每天给予相应药物灌胃处理,观察其体重、生存状态,第21天时,计算大鼠生存率,采用HE染色法观察胶质瘤脑组织的病理变化,免疫组织化学法检测大鼠脑肿瘤浸润区胶质纤维酸性蛋白(glial fibril-lary acidic protein,GFAP)和半胱天冬酶(cysteinyl aspartate specific proteinase,Caspase)-3的表达,Western blot法检测脑肿瘤浸润区血管内皮生长因子(vascular endothelial growth factor,VEGF)、ZO-1、P-gp及Caveolin-1蛋白表达的变化.结果 与模型组相比,替莫唑胺、扶正抑瘤方及联合用药组肿瘤浸润区GFAP和Caspase-3的表达显著上升(P<0.01),ZO-1和P-gp的表达显著下降(P<0.05,P<0.01);与模型组相比,扶正抑瘤方和联合用药组VEGF的表达显著下降(P<0.05,P<0.01),与模型组相比,联合用药组Caveolin-1的表达明显下降(P<0.05).结论 扶正抑瘤方联合替莫唑胺对脑胶质瘤具有明显的防治作用,可促进胶质瘤细胞的分化和凋亡,抑制血管新生,开放血肿瘤屏障,限制肿瘤生长.%Objective To investigate the effect of Fuzheng Yiliu( FZYL) formula combined with temozolomide ( TMZ) on tumor growth and zonula odluden ( ZO-1) , P-glycoprotein ( P-gp) and Caveolin-1 in C6 glioma rats. Methods 50 Wistar male rats were randomly divided into 5 groups, sham-operated group (Sham),model group (MO),FZYL formula group,TMZ group,FZYL+TMZ group. C6 rat glioma model was established by stereotaxic method, the rats in treatment groups were administrated intragastrically with corresponding medicines and the weight and living state were recorded. At twenty-first days, the sur-vival rate was calculated and the pathological changes of brain tissue were observed by HE staining. The protein expression of glial fibrillary acidic protein ( GFAP ) , cysteinyl aspartate specific proteinase-3 ( Caspase-3) were measured by immunohistochemical method and the expression of vascular endothelial growth factor ( VEGF) ,ZO-1,P-gp and Caveolin-1 in invasion area of brain tumor were tested by Western blot method.Results Compared with model group,the expression levels of GFAP and Caspase-3 were obvi-ously increased while the levels of ZO-1 and P-gp were obviously decreased in TMZ group,FZYL group and FZYL+TMZ group (P<0.05,P<0.01), the levels of VEGF were obviously decreased in FZYL group and FZYL+TMZ group (P<0.05,P<0.01),and the levels of Caveolin-1 were obviously decreased in FZYL+TMZ group( P<0.05) . Conclusion FZYL formula combined with TMZ have obvious preventive and thera-peutic effects on glioma, which can promote differentiation and apoptosis of glioma cells, inhibit angiogene-sis, reduce the limitation of the blood brain barrier and inhibite the growth of tumor.