微流控法制备载雷公藤红素脂质体及其抗结肠癌初步研究  

Preparation of Celastrol-loaded Liposomes by Microfluidic Method and Its Preliminary Study on Anti-colon Cancer

作　　者：颜煜雯 张厉梅 朱华泰 王菲[1] 张蓓 雷建都[1] YAN Yuwen;ZHANG Limei;ZHU Huatai;WANG Fei;ZHANG Bei;LEI Jiandu(Engineering Research Center of Forestry Biomass Materials and Energy of Ministry of Education,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学林业生物质材料与能源教育部工程研究中心,北京100083

出　　处：《中国生物工程杂志》2024年第10期18-28,共11页China Biotechnology

基　　金：国家重点研发计划(2019YFB1309703);北京市自然科学基金(2202034)资助项目。

摘　　要：目的:基于微流控法精确控制脂质体粒径和尺寸分布的独特优势,同时为解决雷公藤红素(celastrol,Cel)水溶性差等缺点,制备包载雷公藤红素的脂质体(Cel-LPs),并研究Cel-LPs对结直肠癌的治疗效果。方法:采用微流控法制备Cel-LPs,通过单因素法优化处方和制备条件,对优化得到的Cel-LPs进行形貌、粒径、多分散性指数(polydispersity index,PDI)、Zeta电位、体外释放和稳定性研究;采用CCK-8法检测Cel和Cel-LPs对人结肠癌细胞(HCT116)的毒性;以香豆素6(coumarin6,Cou6)作为荧光物质制备香豆素6脂质体(Cou6-LPs),通过倒置荧光显微镜观察HCT116细胞对Cou6-LPs的摄取行为。结果:优化处方和制备条件,水相与油相流速比为4.5∶1,药物与磷脂质量比为1∶4,胆固醇与磷脂质量比为1∶5;按优化条件制得的Cel-LPs外观呈球形,分布均匀,粒径和PDI分别为(85.02±0.43)nm、0.12±0.01,Zeta电位为(-11.4±0.25)mV,包封率为(95.42±0.12)%;制备的脂质体均一稳定,在体外释放实验中呈现出较好的缓释效果;Cel-LPs对HCT116细胞表现出良好的细胞毒性,抑制其增殖的作用强于Cel,半数抑制浓度(IC_(50))为6.26μg/mL;体外细胞摄取实验表明,HCT116细胞对Cou6-LPs有较好的摄取效果。结论:采用微流控法成功制备了一种稳定、形态均一、分散良好、包封率较高的Cel-LPs,验证了其作为脂质体制备方法的独特特性及广阔应用前景;Cel-LPs可被HCT116细胞较好摄取并缓慢释放,能够有效抑制HCT116细胞增殖,表现出良好的抗肿瘤效果。Objective:Based on the unique advantage of the microfluidic method to precisely control the particle size and size distribution of liposomes in liposome preparation,and in order to overcome the shortcomings such as poor water solubility of Celastrol,Cel-LPs were prepared and the therapeutic effect of Cel-LPs on colorectal cancer was investigated with the aim of providing a safe and effective platform.Methods:Cel-LPs were prepared by the microfluidic method and the formulations were optimized by one-way testing;the morphology,particle size,polydispersity index(PDI),Zeta potential,in vitro release and stability of the optimized formulations of Cel-LPs were investigated;the toxicity of Cel and Cel-LPs on human colon cancer cells(HCT116)was determined by the CCK-8 method;coumarin 6(coumarin 6,Cou6)was used as a fluorescent substance to prepare coumarin 6 liposomes(Cou6-LPs).The uptake behavior of Cou6-LPs by HCT116 cells was investigated by inverted fluorescence microscopy.Results:The optimized formulation was as follows:aqueous phase to oil phase flow rate ratio of 4.5∶1;drug to phospholipid ratio of 1∶4;cholesterol to phospholipid ratio of 1∶5.Cel-LPs prepared according to the optimized prescription had a spherical appearance and uniform distribution,with particle sizes and PDIs of(85.02±0.43)nm and 0.12±0.01,respectively,a Zeta potential of(-11.4±0.25)mV,and an encapsulation rate of(95.42±0.12)%.The prepared liposomes had good stability and showed a good slow release effect in the in vitro release experiments.Cel-LPs showed good cytotoxicity against HCT116 cells,and inhibited their proliferation more than Cel,with a half inhibitory concentration(IC_(50))of 6.26μg/mL.The in vitro cellular uptake experiments showed that HCT116 cells had a better uptake effect of Cou6-LPs in the in vitro cellular uptake experiments.Conclusion:A stable,homogeneous,well-dispersed liposome of tretinoin with a high encapsulation rate was successfully prepared by the microfluidic method,confirming its unique characteristics and

关 键 词：雷公藤红素 脂质体 微流控法 结直肠癌 

分 类 号：Q819[生物学—生物工程]