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目的:制备胰岛素-羧甲基壳聚糖-聚乙二醇纳米粒。方法:利用红外光谱(FTIR)和核磁共振氢谱(~1 H-NMR)对羧甲基壳聚糖-聚乙二醇的结构进行表征,用粒度分析仪测定纳米粒的粒径分布及电位,采用动态透析法考察纳米粒的释药性能,用CCK-8试剂盒检测纳米粒细胞毒性,以糖尿病小鼠为模型,研究纳米粒的降血糖作用。结果:聚乙二醇成功接枝到羧甲基壳聚糖上,包埋胰岛素的纳米粒的平均粒径为(257.5±12.1)nm,Zeta电位为(-15.2±0.3)mV,负载胰岛素的羧甲基壳聚糖-聚乙二醇纳米粒在中性释放介质中,5 h内胰岛素的释放速度较快,之后8 h趋于平稳,胰岛素的累计释放量可达到80%,CCK-8试剂盒显示纳米粒对L929细胞基本无细胞毒性,50 U·kg~(-1)的纳米粒溶液经灌胃给药后,血糖浓度明显降低。结论:胰岛素-羧甲基壳聚糖-聚乙二醇纳米粒基本无毒性,具有良好的生物相容性,对糖尿病小鼠有效发挥降血糖作用。
Objective: To prepare insulin-carboxymethyl chitosan-polyethylene glycol nanoparticles. Methods: The structure of carboxymethyl chitosan-PEG was characterized by FTIR and 1H-NMR. The particle size distribution and potential of nanoparticles The drug release properties of nanoparticles were investigated by dynamic dialysis. The cytotoxicity of nanoparticles was detected by CCK-8 kit. The model of diabetic mice was used to study the hypoglycemic effect of nanoparticles. RESULTS: Polyethylene glycol was successfully grafted onto carboxymethyl chitosan. The average diameter of nanoparticles coated with insulin was (257.5 ± 12.1) nm and the zeta potential was (-15.2 ± 0.3) mV. The loaded insulin In neutral release medium, carboxymethyl chitosan-PEGylated nanoparticles release insulin fast within 5 h, then steady at 8 h, and the cumulative release of insulin can reach 80%. CCK-8 The kit showed that the nanoparticles had no cytotoxicity on L929 cells. The concentration of 50 U · kg ~ (-1) nanoparticles decreased significantly after intragastric administration. CONCLUSION: Insulin-carboxymethyl chitosan-PEGylated nano-particles are basically non-toxic, have good biocompatibility and effectively exert hypoglycemic effect on diabetic mice.