健脾消癌方对人大肠癌裸鼠皮下移植瘤微血管生成的影响

Effects of Jianpi Xiaoai Formula on microvessel density generation of subcutaneous transplantation tumor in human colorectal cancer nude mice

目的:研究健脾消癌方对人肠癌裸鼠皮下移植瘤微血管生成的抑制作用。方法:通过在裸鼠腋下接种人肠癌HCT116细胞建立裸鼠人肠癌移植瘤模型。60只荷瘤裸鼠随机分为模型组、5-氟尿嘧啶(5-Fu)组及健脾消癌方高、中、低剂量组,每组12只,各组分别给予0.9%氯化钠溶液、5-Fu(30mg/kg,1次/d)及不同浓度的健脾消癌方(高、中、低剂量)干预。给药21d后,每组随机处死6只裸鼠,摘除瘤体并眼球取血,测量瘤体重量和体积,ELISA法检测裸鼠血清中血管内皮生长因子(VEGF)的表达、免疫组化检测瘤体微血管密度(MVD);每组剩余裸鼠观察荷瘤鼠生存时间。结果:与模型组比较,药物干预组的瘤体重量均有不同程度减轻(P<0.01)、荷瘤鼠中位生存时间显著延长(P<0.01),其中5-Fu组和健脾消癌方高、中剂量组在瘤体重量、抑瘤率及延长荷瘤鼠中位生存时间方面均优于健脾消癌方低剂量组(P<0.01),且健脾消癌方高剂量组与5-Fu组的作用差异无统计学意义。同时,健脾消癌方能显著下调裸鼠血清中VEGF表达水平(P<0.01);免疫组化结果显示,健脾消癌方以浓度依赖性方式下调肿瘤组织MVD表达,与模型组比较差异显著(P<0.01)。结论:健脾消癌方能够抑制裸鼠人大肠癌移植瘤生长、延长生存期,其作用可能和降低血清VEGF表达及肿瘤MVD有关。

Objective: To study the inhibition effect of Jianpi Xiaoai Formula on human colorectal cancer nude mice subcutaneously transplanted tumor microvascular generation. Methods: Tumor cell line HCT116 was inoculated subcutaneously into nude mice axilla to establish human colorectal cancer xenograft model, 60 nude mice were randomly divided into model group, 5-Fu group, high, medium and low dose Jianpi Xiaoai Formula group, each group were given normal saline, 5-Fu and different concentrations of Jianpi Xiaoai Formula(high, medium and low dose) respectively. After 21 days, 6 rats were sacrificed each group randomly, the tumor were removed and eyeball blood were collected, the body weight and volume of tumor were observed and the contents of VEGF in blood serum and tumor microvessel density(MVD) were measured. The survival time of tumor bearing mice was observed in each group of remaining nude mice. Results: Compared with the model group, the tumor weight in drug intervention groups all alleviated in different degree(P<0.01), the median survival time of tumor bearing mice was significantly prolonged(P<0.01), the 5-Fu group and the high and medium dose Jianpi Xiaoai Formula group were better than the low dose Jianpi Xiaoai Formula group in tumor weight, inhibition rate and prolonging nude mice median survival time(P<0.01), and there was no significant difference between Jianpi Xiaoai Formula high dose group and 5-Fu group. At the same time, compared with the model group, Jianpi Xiaoai Formula can significantly reduce the contents of VEGF in blood serum(P<0.01); immunohistochemistry showed that Jianpi Xiaoai Formula can reduce the expression of MVD in tumor tissue with a concentration dependent manner(P<0.01). Conclusion: Jianpi Xiaoai Formula can inhibit tumor growth and prolong survival time of tumor bearing nude mice, and the effect may be related to the decrease of serum VEGF expression and tumor microvessel density.