EGCG-CS-PAA纳米粒的制备及其生物活性研究
发布时间：2018-10-19 10:41
【摘要】：EGCG陛质很不稳定,易受光、氧、温度、pH、金属离子等外界因素的影响从而严重限制了其应用的前景。在参考前人研究的基础上,发现壳聚糖纳米技术能较好的保护EGCG免于遭受恶劣环境的破坏。壳聚糖纳米粒包埋EGCG后,由于纳米粒的优越特性,使得EGCG生物功效得到提高,同时EGCG的释放也变缓,从而提高了EGCG的利用效率。 在本研究中,利用壳聚糖CS和聚天冬氨酸PAA之间离子交联制备CS-PAA纳米粒载体,以此装载EGCG,然后对EGCG-CS-PAA纳米粒的稳定性和生物活性等做了相关研究。研究发现,不同的制备参数能显著影响纳米粒的粒径、分散指数和Zeta电位等特性。当质量比(CS/PAA)为1.0、pH为3.5、反应时间为60min、CS分子量为3-5KDa和PAA分子量为30-50KDa时,制备出的壳聚糖纳米粒的粒径和表面电荷等物理特征为最佳。同时,在该制备参数下制备出的EGCG-CS-PAA纳米粒的包封率达到25%,载药量为344.1mg/kg,且其粒径比未包埋EGCG的CS-PAA纳米粒大些。 EGCG-CS-PAA纳米粒的体外释放研究表明在模拟胃环境(pH=2.5-4.0)下从纳米粒中释放出来的EGCG含量较少；在模拟肠环境(pH=6.0-7.4)下,纳米颗粒容易发生降解从而迅速释放EGCG。通过采用FRAP和DPPH法对EGCG-CS-PAA纳米粒的抗氧化活性进行分析,两者都显示纳米体系对EGCG的抗氧化活性具有良好的保护作用。同时EGCG-CS-PAA纳米体系能在高温、碱性等恶劣环境下对EGCG起到较好的保护作用。 白兔体内试验结果显示,相比于自由的EGCG, EGCG-CS-PAA纳米粒在预防白兔动脉粥样硬化及其降血脂功效方面得到明显的提高。在EGCG纳米粒小鼠急性试验中,在实验最大剂量(60mL/kg)范围内,雌雄小鼠均未出现死亡及异常情况,且给药后对小鼠的体重也无显著影响。同时,细胞毒性试验结果也表明EGCG-CS-PAA纳米粒及所制备的材料均为无毒安全的。 本研究结果表明,CS-PAA纳米体系可以作为如EGCG等不稳定功能性药物的载体,有效保护其生物稳定性,为EGCG在医药食品等领域的应用开拓新的途径。
[Abstract]:EGCG is unstable and vulnerable to light, oxygen, temperature, pH, metal ions and other external factors, which seriously limits its application prospects. On the basis of previous studies, it was found that chitosan nanotechnology could protect EGCG from severe environmental damage. After EGCG was encapsulated by chitosan nanoparticles, because of the superior properties of nanoparticles, the biological efficacy of EGCG was improved, and the release of EGCG was slowed down, thus improving the utilization efficiency of EGCG. In this study, the carrier of CS-PAA nanoparticles was prepared by ion crosslinking between chitosan CS and polyaspartic acid PAA, and then the stability and bioactivity of EGCG-CS-PAA nanoparticles were studied by loading EGCG,. It is found that different preparation parameters can significantly affect the particle size, dispersion index and Zeta potential. When the mass ratio (CS/PAA) is 1.0 渭 g pH is 3.5, the reaction time is 60 min CS molecular weight is 3-5KDa and PAA molecular weight is 30-50KDa, the size and surface charge of the prepared chitosan nanoparticles are the best. meanwhile The encapsulation efficiency of EGCG-CS-PAA nanoparticles prepared under this preparation parameter is 25mg / kg, the drug loading is 344.1 mg / kg, and its diameter is larger than that of CS-PAA nanoparticles without EGCG. The in vitro release study of EGCG-CS-PAA nanoparticles shows that under simulated gastric environment (pH=2.5-4.0), The content of EGCG released from nanoparticles was less. Under simulated intestinal environment (pH=6.0-7.4), nanoparticles are liable to degrade and release EGCG. rapidly. The antioxidant activity of EGCG-CS-PAA nanoparticles was analyzed by FRAP and DPPH. Both of them showed that the nanoparticles had good protective effect on the antioxidant activity of EGCG. At the same time, EGCG-CS-PAA nanosystem can protect EGCG in high temperature and alkaline environment. The results showed that compared with the free EGCG, EGCG-CS-PAA nanoparticles, the effect of preventing atherosclerosis and lowering blood lipids in white rabbits was significantly improved. In the acute test of EGCG nanoparticles mice, in the range of maximum experimental dose (60mL/kg), there was no death and abnormality in both male and female mice, and there was no significant effect on the body weight of mice after administration of the drug. At the same time, the results of cytotoxicity test also showed that the EGCG-CS-PAA nanoparticles and the prepared materials were nontoxic and safe. The results show that CS-PAA nanosystem can be used as a carrier of unstable functional drugs such as EGCG, protect its biological stability effectively, and open up a new way for the application of EGCG in the field of medicine and food.
【学位授予单位】：浙江工商大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TQ460.1;TB383.1
本文编号：2280886
[Abstract]:EGCG is unstable and vulnerable to light, oxygen, temperature, pH, metal ions and other external factors, which seriously limits its application prospects. On the basis of previous studies, it was found that chitosan nanotechnology could protect EGCG from severe environmental damage. After EGCG was encapsulated by chitosan nanoparticles, because of the superior properties of nanoparticles, the biological efficacy of EGCG was improved, and the release of EGCG was slowed down, thus improving the utilization efficiency of EGCG. In this study, the carrier of CS-PAA nanoparticles was prepared by ion crosslinking between chitosan CS and polyaspartic acid PAA, and then the stability and bioactivity of EGCG-CS-PAA nanoparticles were studied by loading EGCG,. It is found that different preparation parameters can significantly affect the particle size, dispersion index and Zeta potential. When the mass ratio (CS/PAA) is 1.0 渭 g pH is 3.5, the reaction time is 60 min CS molecular weight is 3-5KDa and PAA molecular weight is 30-50KDa, the size and surface charge of the prepared chitosan nanoparticles are the best. meanwhile The encapsulation efficiency of EGCG-CS-PAA nanoparticles prepared under this preparation parameter is 25mg / kg, the drug loading is 344.1 mg / kg, and its diameter is larger than that of CS-PAA nanoparticles without EGCG. The in vitro release study of EGCG-CS-PAA nanoparticles shows that under simulated gastric environment (pH=2.5-4.0), The content of EGCG released from nanoparticles was less. Under simulated intestinal environment (pH=6.0-7.4), nanoparticles are liable to degrade and release EGCG. rapidly. The antioxidant activity of EGCG-CS-PAA nanoparticles was analyzed by FRAP and DPPH. Both of them showed that the nanoparticles had good protective effect on the antioxidant activity of EGCG. At the same time, EGCG-CS-PAA nanosystem can protect EGCG in high temperature and alkaline environment. The results showed that compared with the free EGCG, EGCG-CS-PAA nanoparticles, the effect of preventing atherosclerosis and lowering blood lipids in white rabbits was significantly improved. In the acute test of EGCG nanoparticles mice, in the range of maximum experimental dose (60mL/kg), there was no death and abnormality in both male and female mice, and there was no significant effect on the body weight of mice after administration of the drug. At the same time, the results of cytotoxicity test also showed that the EGCG-CS-PAA nanoparticles and the prepared materials were nontoxic and safe. The results show that CS-PAA nanosystem can be used as a carrier of unstable functional drugs such as EGCG, protect its biological stability effectively, and open up a new way for the application of EGCG in the field of medicine and food.
【学位授予单位】：浙江工商大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TQ460.1;TB383.1
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