水飞蓟素脂质体的制备、质量评价及其对HaCaT细胞增殖和摄取的影响  

Preparation and quality evaluation of silymarin liposomes and their effects on proliferation and uptake of HaCaT cells

作　　者：陈江丽 邱智东[1] 金媛媛 周栩妍 李弘鹏 刘达[1] 金叶 CHEN Jiang-li;QIU Zhi-dong;JIN Yuan-yuan;ZHOU Xu-yan;LI Hong-peng;LIU Da;JIN Ye(School of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学药学院,吉林长春130117

出　　处：《中草药》2022年第12期3614-3624,共11页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(82003985);吉林省教育厅科学技术研究项目(JJKH20210995KJ);吉林省科技发展计划项目(20210204013YY)。

摘　　要：目的 制备水飞蓟素脂质体(silymarin liposomes,SM-L)并进行质量评价,考察SM-L对人永生化角质形成细胞(human immortal keratinocyte line,HaCaT)增殖和摄取的影响。方法 采用乙醇注入法制备SM-L,以包封率和ζ电位为考察指标,Box-Behnken响应曲面设计实验优化SM-L处方和制备工艺,并对SM-L进行质量评价;采用噻唑蓝(MTT)法考察SM-L对HaCaT细胞增殖的影响,细胞划痕实验考察SM-L对HaCaT细胞水平迁移能力的影响,共聚焦显微镜观察并以流式细胞术考察HaCaT细胞对SM-L的摄取情况。结果 优化的SM-L制备工艺为大豆磷脂、水飞蓟素和胆固醇的质量比为20∶7.94∶6.21,无水乙醇和水的最佳比例为1∶10。SM-L包封率为(85.44±0.87)%,平均粒径为(133.80±2.13)nm,多分散指数为0.134±0.020,ζ电位为(-14.1±0.3)m V,稳定性良好,36 h皮肤透过率为(14.58±6.37)%,皮肤滞留率为(65.80±4.04)%。细胞实验证明一定质量浓度范围的SM-L能促进HaCaT细胞增殖,SM-L相比水飞蓟素对HaCaT细胞的水平迁移促进作用有明显提高,并且SM-L能够有效提高HaCaT细胞对水飞蓟素的摄取。结论 制备所得的SM-L粒径均一,性质稳定,透皮性能好,并且可以促进HaCaT细胞增殖和对水飞蓟素的摄取,有很好的角质修复作用。Objective Silymarin liposomes(SM-L) were prepared and evaluated to investigate its effect on the uptake and migration of HaCaT cells. Methods SM-L was prepared by ethanol injection method. Taking the entrapment efficiency and ζ potential as the indexes, the Box-Behnken response surface design was used to optimize the best formulation and process of SM-L, and the quality of SM-L was evaluated;MTT method was used to investigate the effect of SM-L on the proliferation of HaCaT cells, cell scratch test was used to investigate the effect of SM-L on the horizontal migration ability of HaCaT cells, confocal microscope and flow cytometry were used to investigate the uptake of SM-L by HaCaT cells. Results The best preparation process of SM-L was as follows: the mass ratio of soybean phospholipid, silymarin(SM-L) and cholesterol was 20:7.94:6.21, and the optimum ratio of organic phase and aqueous phase was 1:10. The entrapment efficiency of SM-L was(85.44 ± 0.87)%, the particle size was(133.80 ± 2.13) nm, the polydispersity index was 0.134 ± 0.020, the ζ potential was(-14.1 ± 0.3) mV, the skin transdermal rate was(14.58 ± 6.37)%, and skin retention rate was(65.80 ± 4.04)% within 36 h, and SM-L had good stability. Cell experiments showed that SM-L in a certain concentration range could promote the proliferation of HaCaT cells. Compared with silymarin, SM-L could significantly improve the horizontal migration of HaCaT cells, and SM-L could effectively improve the uptake of silymarin by HaCaT cells. Conclusion The prepared SM-L has uniform particle size, good stability and transdermal performance, can promote HaCaT cell proliferation and silymarin uptake, and has a good role in cuticle repair.

关 键 词：水飞蓟素 脂质体 质量评价 细胞增殖 细胞摄取 乙醇注入法 

分 类 号：R283.6[医药卫生—中药学]