【作者】 蒋蕾；

【导师】 霍介格；

【作者基本信息】 南京中医药大学 ， 中医内科学， 2012， 硕士

【摘要】 目的1.观察小分子血管破坏药物Combretastatin A4Phosphate (CA4P)与十全大补汤合用对小鼠H22肝癌移植瘤生长的抑制作用,并初步探讨其作用机制。2.观察CA4P联合磁热疗对小鼠H22肝癌移植瘤生长的抑制作用。方法1.建立小鼠H22肝癌移植瘤模型,将成瘤小鼠随机分成7组：Ⅰ组：CA4P单药组,Ⅱ组：十全大补汤低剂量组,Ⅲ组：十全大补汤高剂量组,Ⅳ组：CA4P+十全大补汤低剂量组,Ⅴ组：CA4P+十全大补汤高剂量组,Ⅵ组：空白对照组。用药期间每3天测量移植瘤长短经,计算移植瘤体积,绘制肿瘤生长曲线,用药28天后颈椎脱臼法处死小鼠,剥离皮下移植瘤称重,计算抑瘤率,评价联合用药效应；HE染色,镜下观察移植瘤组织形态变化；MRI检测肿瘤坏死率；计算免疫脏器指数,流式细胞检测小鼠外周血中CD3+、CD4+、CD8+细胞比率,ELISA试剂盒检测外周血中IL-6、IFN-γ水平,观察联合用药对荷瘤鼠免疫功能的影响。2.建立小鼠H22肝癌移植瘤模型,将小鼠随机分成4组：NS对照组(NS),磁流体热疗组(MFH), CA4P单药组(CA), CA4P联合磁热疗组(CMFH)。用药期间每3天测量移植瘤长短经,计算移植瘤体积,绘制肿瘤生长曲线,用药14天后颈椎脱臼法处死小鼠,剥离皮下移植瘤称重,计算抑瘤率,评价联合用药效应；HE染色,镜下观察移植瘤组织形态变化。结果1.(1)CA4P联合十全大补汤对移植瘤增长无明显抑制作用,CA4P单药组、十全大补汤低剂量组、十全大补汤高剂量组、CA4P+低剂量组、CA4P+高剂量组各组抑瘤率分别为：4.43%、11.61%、20.46%、2.22%、6.62%。(2)各给药组移植瘤肿瘤坏死率明显增高,十全大补汤低、高剂量联合CA4P较CA4P单药组肿瘤坏死面积增大。(3)各给药组对荷瘤鼠免疫脏器指数无影响,CA4P联合十全大补汤可升高荷瘤鼠外周血CD3+、CD4+、CD8+细胞比率(P<0.05),CA4P可使小鼠血清中IL-6、IFN-γ含量明显升高(P<0.01),联合十全大补汤抑制该调节作用。2.CA4P、磁热疗可抑制移植瘤生长,CA4P联合磁热疗组抑瘤作用明显增强(P<0.01),CA、MFH、CMFH各组抑瘤率分别为：42.3%、63.3%、82.54%。HE染色示各治疗组肿瘤坏死区增大,CMFH组肿瘤坏死最为明显。结论1.(1)CA4P联合十全大补汤可促进移植瘤组织坏死,但对移植瘤增长无明显抑制作用；(2)CA4P联合十全大补汤可促进荷瘤鼠T细胞增殖活化,十全大补汤可拮抗CA4P促进细胞因子分泌的作用。2.CA4P联合磁热疗可明显抑制小鼠H22肝癌移植瘤增长。更多还原

【Abstract】 Objective1.To investigate the effect of a Low-molecular-weight vascular-disrupting agent Combretastatin A4phosphate (CA4P) combination with Shiquandabu Decoction on growth of H22hepatoma xenograft mice and its mechanism.2.To investigate the effect of CA4P combination with Magnetic fluid Hyperthermia(MFH) on growth of H22hepatoma xenograft mice.Methods1.The liver cancer xenograft mode in mice was established H22cell.Seventy eight mice with liver cancer xenograft were randomly divided into six groups:I CA4P group; II Shiquandabu Decoction low-dose group; III Shiquandabu Decoction high-dose group; IV CA4P+Shiquandabu Decoction low-dose group; V CA4P+Shiquandabu Decoction high-dose group; VIcontrol group.The lenghth and width of xenograft were measured by vernier caliper at every three days during the therapy time.The volume of the xenograft was calculated,and xenograft grouth curves were ploted out.After28days,the tumor necrosis ratio was measured by MRI,the weight of primary subcutaneous xenograft and the inhibitory rate of weight of xenograft were examined after all mice sacrificed.Recipe blood from vein of eyeball centrifuging.The T cell subsets in peripheral blood were detected by FCM.The contents of IL-6and IFN-y of blood serum were detected by ELISA.The pathology form of xenograft were observed under microscop.2.The hepatoma-xenograft mice were constructed as moder,divided into four groups randomly:control group(NS),CA4P group(CA), Magnetic fluid Hyperthermia group(MFH) and CA4P+MFH group (CMFH). The lenghth and width of xenograft were measured by vernier caliper at every three days during the therapy time.The volume of the xenograft was calculated,and xenograft grouth curves were ploted out.The weight of primary subcutaneous xenograft and the inhibitory rate of weight of xenograft were examined at14days after the last hyperthermia after all mice sacrificed. The pathology form of xenograft were observed under microscop.Results1.(1)Comparing the tumor growth of all groups with that of control group,it hadn’t remarkable difference(P>0.05). The inhibitory rate of weight of subcutaneous xenograft in CA4P group,Shiquandabu Decoction low-dose group,Shiquandabu Decoction high-dose group,CA4P+low-dose group,CA4P+high-dose group,control group were respectively:4.43%、11.61%、20.46%、2.22%、6.62%.(2) In every treatment group the tumor necrosis ratio was increased, CA4P combination with Shiquandabu Decoction showed larger necrosis areas than CA4P group.(3) CA4P combination with Shiquandabu Decoction can increase CD3+、 CD4+、 CD8+T cell ratio in peripheral blood. CA4P can promote markedly production of IL-6and IFN-y, Shiquandabu Decoction can produce protective effect.2.In every treatment group xenograft growth was suppressed significantly.CA4P plus MFH group showed enhancement efficacy in anti-tumor growth(P<0.01).The inhibitory rate of weight of subcutaneous xenograft were respectively423%、63.3%、82.54%。 Pathological examination showed much larger necrosis areas in every treatment group.CA4P plus MFH showed the most obvious changes.Conclusions1.(1) CA4P combination with Shiquandabu Decoction can aggravate tissue necrosis in the xenograft,however,it can not inhibit the xenograft growth.(2) CA4P combination with Shiquandabu Decoction can promote the proliferation and activation of T cells, CA4P can significantly promote cytokines production, Shiquandabu Decoction exerts a protective action against this effect.2. CA4P combination with MFH can significantly inhibit the growth of H22hepatoma-xenograft in mice.更多还原