上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (6): 862-.doi: 10.3969/j.issn.1674-8115.2011.06.041

• 技术与方法 • 上一篇    下一篇

缓释纳米尿激酶的制备

金海将, 张 皓, 张柏根, 孙敏莉   

  1. 上海交通大学 医学院附属仁济医院血管外科, 上海 200001
  • 出版日期:2011-06-28 发布日期:2011-06-27
  • 通讯作者: 张 皓, 电子信箱: chlzhcx@163.com。
  • 作者简介:金海将（1984—）, 男, 博士生；电子信箱: jinhaijiang521@163.com。
  • 基金资助:

    上海市科委课题（0852nm05100）

Preparation of sustained-release urokinase nanoparticles

JIN Hai-jiang, ZHANG Hao, ZHANG Bai-gen, SUN Min-li   

  1. Department of Vascular Surgery, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200001, China
  • Online:2011-06-28 Published:2011-06-27
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 0852nm05100

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摘要：

目的 制备包载尿激酶的水溶性壳聚糖(WSC)纳米粒子，观察纳米尿激酶的活性。方法 采用离子交联法制备包载尿激酶的WSC纳米粒子；检测尿激酶纳米粒子的包封率，分析WSC与三聚磷酸钠质量比、尿激酶浓度、磁力搅拌速度和时间、超声时间和功率等条件对包封率的影响，获得包封率最高的制备条件；粒径仪测量尿激酶纳米粒子粒径；超滤离心法纯化尿激酶纳米粒子，注入新西兰兔体内(纳米尿激酶组)，于不同时间点取血测定尿激酶活性,以注射单纯尿激酶的动物作为对照组。结果 室温下，将三聚磷酸钠溶液（0.6 g/L）滴加入包含尿激酶（1 mg/10 mL)的WSC溶液（1.0 g/L），磁力搅拌速度800 r/min，滴加完毕后继续搅拌60 min，冰浴条件下超声30 s(功率10 W)，成功制备出包封率为94.8%的尿激酶纳米粒子；对照组血尿激酶活性达到峰值后随着时间的延长逐渐降低，纳米尿激酶组血尿激酶活性达到峰值后仍保持较高水平。结论 成功制备了包载尿激酶的WSC纳米粒子，纳米尿激酶具有缓释功能并保持了尿激酶的活性。

关键词: 水溶性壳聚糖, 尿激酶纳米粒子, 包封率, 离子交联法

Abstract:

Objective To prepare urokinase water soluble chitosan (WSC) nanoparticles, and investigate its activity. Methods Urokinase WSC nanoparticles were prepared by ionic crosslinking method, and the encapsulation efficiency of urokinase nanoparticles was measured. The effects of quality ratio of WSC to tripolyphosphate, concentration of urokinase, time and velocity of magnetic stirring, and time and power of ultrasound on encapsulation efficiency were determined, and the conditions for the highest encapsulation efficiency were obtained. The diameters of urokinase nanoparticles were measured by particle size instrument. Urokinase nanoparticles were purified by centrifugal ultrafiltration, and were injected into New Zealand rabbits (urokinase nanoparticles group). Blood samples were taken from rabbits at different time points, and the activity of urokinase nanoparticles was examined. Rabbits injected with single urokinase were served as control group. Results Under room temperature, when tripolyphosphate solution (0.6 g/L) was dropped into WSC solution (1.0 g/L) containing urokinase (1 mg/10 mL), urokinase nanoparticles with the highest encapsulation efficiency (94.8%) were obtained with the time and velocity of stirring of 60 min and 800 r/min respectively, and the time and power of ultrasound in ice bath of 30 s and 10 W respectively. The serum activity of urokinase gradually decreased with time after reaching the peak in control group, while that maintained at a higher level after reaching the peak in urokinase nanoparticles group. Conclusion Urokinase WSC nanoparticles have been successfully prepared, which possess not only sustained-release function, but also activity of urokinase.

Key words: water soluble chitosan, urokinase nanoparticles, encapsulation efficiency, ionic crosslinking method