半乳糖基壳聚糖5氟尿嘧啶纳米粒的制备及其抑制小鼠结肠癌的实验研究

目的:半乳糖基壳聚糖纳米材料与5-氟尿嘧啶(5-FU)合成5-FU纳米粒,并观察其在体内抑制小鼠结肠癌的疗效.方法:将体外培养的C26细胞接种小鼠肝左叶,成瘤后,分别用0.9％氯化钠液、空纳米粒、5-FU和5-FU纳米粒行尾静脉注射,观察5-FU纳米粒的抗肿瘤效果.用酶联免疫吸附试验法检测血清γ干扰素(IFN-γ)和白细胞介素-2(IL-2)含量,用四甲基偶氮唑盐(MTT)比色法测定小鼠脾自然杀伤细胞(NK细胞)和细胞毒性T细胞(CTL细胞)活性.结果:与0.9％氯化钠液组及纳米粒组相比,5-FU组和5-FU纳米粒组的瘤质量显著降低(P＜0.01),尤其5-FU纳米粒组降低更显著(P＜0.01).与其他各组比较,5-FU组小鼠,外周血IFN-γ和IL-2含量均显著降低(P＜0.01),5-FU纳米粒组与0.9％氯化钠液组比较差异无统计学意义(P＞0.05).5-FU组的脾NK细胞和CTL细胞杀伤活性较其他各组显著降低(P＜0.01),5-FU纳米粒组与0.9％氯化钠液组比较未见降低(P＞0.05),纳米粒组NK和CTL杀伤活性0.9％氯化钠液组增高(P＜0.05).结论:5-FU纳米粒治疗结肠癌模型,具有明显抗肿瘤效应,纳米粒材料具有提高机体免疫功能,减低5-FU对机体免疫的抑制作用.

Preparation of 5-Fluorouracil Nanoparticles of Galactosylated Chitosan and Its Inhibition of the Colon Cancer in Mice

Objective：The 5-fluorouracil（5-Fu） nanoparticles of galactosylated chitosan was synthesized and the efficacy were observed in colon cancer model in vivo with 5-fluorouracil nanoparticles of galactosylated chitosan.Methods：C26 cells were cultured left lobe of the liver in mice,after tumor formation,models were treated with normal saline in NS group,empty nanoparticles in NP group,5-Fu nanoparticles in 5-Fu NP group and 5-Fu in 5-Fu group,respectively,via tail vein injection.Tumor weight were measured in the end of this experiment.The serum interferon-gamma（IFN-y） and interleukin-2（IL-2） were detected by enzyme-linked immunospot-based assays（ELISA）,and natural killer（NK） cells and cytotoxic T lymphocytes（CTL） activity of mouse spleen were measured by the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium（MTT） colorimetric assay. Results：Compared with NS group and NP group,the tumor weight in 5-Fu group and group was significantly lighter（P 0.01）,the tumor weight decreased more obvious in 5-Fu NP group（P〈0.01）.The levels of IFN-y and IL-2 of liver metastases of colon cancer model in mice decreased obviously in 5-Fu group than those in other groups（P〈0.01）,however,the decrease of IFN-γand IL-2 in 5-Fu NP group was not obvious（P 〉0.05）,compared with the saline group,while the levels in NP group was significantly higher than those in NS group（P〈0.01）.The killing activity of NK and CTL in 5-Fu group was significantly lower than that in other groups（P〈0.01）,however,the killing activity of NK and CTL in NP group increased compared with NS group（P〈0.05）,and 5-Fu NP group was no lower compared with NS group（P〉 0.05）.Conclusions： The 5-Fu nanoparticles have significant anti-tumor effect in colon cancer model,nanoparticle materials can stimulate immune function of mice and reduced the side effects of 5-Fu in immune suppression.