健脾解毒方对人胃癌裸鼠皮下移植瘤微血管生成的影响

Inhibitory Effects of “Jianpi Jiedu Recipe” on Angiogenesis in Subcutaneous Xenografts of Gastric Cancer in Mice

目的:探讨健脾解毒方对人胃癌裸鼠皮下移植瘤微血管生成的抑制作用。方法:通过接种人胃癌MKN45细胞建立裸鼠人胃癌移植瘤模型。将60只荷瘤裸鼠随机分为模型组、5-氟尿嘧啶(5-Fu)组及健脾解毒方低、中、高剂量组,每组12只,各组分别给予生理盐水、5-Fu(30 mg/kg,ip×5,qd)及不同浓度的健脾解毒方醇提物(250、500、1 000 mg·kg^(-1)·d^(-1),21 d)干预。给药21 d后,各组随机处死6只,眼球取血并摘除瘤体,测量瘤体大小和质量,并用免疫组织化学法检测瘤体的微血管密度(MVD)、ELISA法检测血清中血管内皮生长因子(VEGF)的表达;剩余6只观察荷瘤鼠生存时间。结果:与模型组比较,各药物干预组的瘤体质量均有不同程度减轻(P<0.01)、荷瘤鼠中位生存时间明显延长(P<0.01),其中健脾解毒方中、高剂量组和5-Fu组在瘤体质量、抑瘤率及荷瘤鼠中位生存时间、生命延长率方面均优于健脾解毒方低剂量组(P<0.01),且健脾解毒方高剂量组与5-Fu组的作用均无统计学差异(P>0.05)。免疫组化结果显示,健脾解毒方能够以剂量依赖性方式下调肿瘤MVD表达,与模型组比较差异显著(P<0.01);同时,健脾解毒方能显著下调裸鼠血清中VEGF表达水平(P<0.01)。结论:健脾解毒方能够抑制裸鼠人胃癌移植瘤生长、延长生存期,并对血清VEGF表达及肿瘤微血管生成有抑制作用。

Objective： To investigate the effects of ＂Jianpi Jiedu Recipe＂on angiogenesis in subcutaneous xenografts of gastric cancer in nude mice. Methods： Colorectal cancer model in nude mice bearing human gastric carcinoma MKN45 cells were established,and then 60 nude mice were randomly divided into 5 groups： model control group,low-dose（ 250 mg·kg^（-1）·d^（-1））,middle-dose（ 500 mg·kg^（-1）·d^（-1））,high-dose＂Jianpi Jiedu Recipe＂group（ 1000 mg·kg^（-1）·d^（-1））,and 5-Fu group（ 30 mg / kg,ip × 5,qd）. Six mice in each group were sacrificed at day 21 and tumor weights were measured; the remaining mice（ 6 mice in each group） were maintained longer for survival studies. The expression of microvessel density（ MVD） was detected by immunohistochemical method; vascular endothelial growth factor（ VEGF） level was assayed by enzyme-linked immunosorbent assay（ ELISA）. Results： Compared with the model control group,tumor weight decreased in the low-,middle-,high-dose of＂Jianpi Jiedu Recipe＂and 5-FU groups（ P〈0. 01）; tumor weight and tumor-inhibiting rate in the middle-,high-dose＂Jianpi Jiedu Recipe ＂groups and 5-Fu group were better than those in low-dose ＂Jianpi Jiedu Recipe＂group（ P〈0. 01）; there were no significant differences between high-dose ＂Jianpi Jiedu Recipe＂group and 5-Fu group（ P〈0. 05）. Compared with the model group,median survival time in the low-,middle-,high-dose ＂Jianpi Jiedu Recipe＂groups and 5-Fu group were prolonged（ P〈0. 01）; median survival time and Life span in the middle-,high-dose＂Jianpi Jiedu Recipe＂groups and 5-Fu group were better than those in low-dose＂Jianpi Jiedu Recipe＂group（ P〈0. 01）.The immunohistochemical results showed that ＂Jianpi Jiedu Recipe＂could down-regulate MVD in a dose-dependent manner compared with model group（ P〈0. 01）. Meanwhile,＂Jianpi Jiedu Recipe＂significantly reduced the serum level of VEGF expression in nude mice（ P〈0. 01）. Conclusion： ＂Jianpi Jiedu Recipe＂can significantly inhibit tumor angiogenesis and growth of gastric cancer xenografts in mice,and extend survival via inhibition of VEGF and MVD.