组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2014, Vol. 10 ›› Issue (2): 77-81.doi: 10.3969/j.issn.1673-0364.2014.02.004

• 论著 • 上一篇    下一篇

5-氟尿嘧啶醇脂体抑制瘢痕成纤维细胞的实验研究

张铮，沃雁，张振，毛小慧，苏薇洁，濮哲铭，张艳，章一新   

  1. 上海交通大学医学院附属第九人民医院整复外科
  • 发布日期:2020-07-23

Inhibitory Effects of Ethosomes Encapsulated with 5-Fluorouracil on Human Hypertrophic Scar Fibroblasts

ZHANG Zheng,WO Yan,ZHAGN Zhen,MAO Xiaohui,SU Weijie,PU Zheming,ZHANG Yan,ZHANG Yixin   

  1. Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine;
  • Published:2020-07-23
  • Contact: 国家自然科学基金(81071569)

摘要： 目的构建纳米级载5-氟尿嘧啶醇脂体(5-FU ES),观察其对瘢痕成纤维细胞的抑制作用。方法采用Touitou法制备5-FU ES悬液并进行质量评价。体外培养增生性瘢痕成纤维细胞,荧光素RhoB标记醇脂体,进行体外透细胞实验,以水醇溶液为对照。激光共聚焦显微镜(CLSM)观察不同作用时间后细胞内荧光分布和强度。CCK-8检测不同浓度5-FU ES对成纤维细胞活性的影响,计算载5-FU ES对成纤维细胞活性的半数抑制浓度(IC50)。CCK-8检测醇脂体对瘢痕成纤维细胞生长的影响。结果制备所得5-FU ES粒径为(87.5±5.4)nm,分散系数为0.151±0.27,包封率为(10.03±2.12)%。CLSM图像显示,醇脂体促进RhoB进入细胞内,可见荧光分布和强度均较水醇溶液组增多增强;经Image-Pro Plus 6.0图像分析软件计算单位面积光密度值,RhoB标记醇脂体组(Rho ES)细胞内荧光光密度显著高于水醇溶液组(Rho HA)(P<0.01)。随着5-FU浓度增加,5-FU HA和5-FU ES对成纤维细胞的生长抑制率逐渐增强;5-FU ES和5-FU HA对成纤维细胞的半数抑制浓度(IC50)分别为(9.2582±1.2329)μg/mL和(18.0352±2.3145)μg/mL,差异具有统计学意义(P<0.05);同时,醇脂体本身对细胞活性无明显抑制作用。结论醇脂体可有效携带5-FU向瘢痕成纤维细胞内递送,促进5-FU对成纤维细胞的活性抑制作用。醇脂体作为经皮给药载体,是一种安全有效的透细胞药物载体。

关键词: 醇脂体, 5-氟尿嘧啶, 增生性瘢痕, 成纤维细胞, 细胞内递送

Abstract: Objective To establish ethosomes encapsulated with 5-Fluorouracil and to explore the inhibitory effects of 5-FU on human hypertrophic scar fibroblasts. Methods Ethosomes encapsulated with 5-Fluorouracil was prepared by Touitou method and was evaluated. Human hypertrophic scar fibroblasts were cultured in vitro, the penetration of the fluorescent probes into fibroblasts was examined by CLSM, and hydroalcoholic solution was set as control group. The effect of ethosomes encapsulating different concentration of 5-FU on fibroblast viability was tested by cell counting kit-8 (CCK-8), and the median inhibitory concentration (IC50) was calculated. The effect of ethosomes on fibroblast viability was tested by CCK-8. Results The particle size of 5-FU encapsulated ethosomes was (87.5 ±5.4) nm, the polydispersion index was 0.151 ±0.27, and the encapsulation efficiency of 5-FU was (10.03 ±2.12)%. CLSM micrographs showed that ethosomes facilitated the penetration of RhoB into the cell, as evident from the high intensity fluorescence. In comparison, when incorporated in hydroalcoholic solution, very weak fluorescence was detected. The optical density (OD) of unit area was calculated through the Image-Pro Plus 6.0 image analysis software, and the OD value of ethosomal group was distinguished higher than hydroalcoholic group (P<0.01). With the increase of the concentration of 5-FU, growth inhibition rate of fibroblast affected by 5-FU HA and 5-FU ES was gradually enhanced. The IC50 of 5-FU encapsulated ethosomes and 5-FU hydroalcoholic solution to fibroblasts were (9.2582±1.2329) μg/mL and (18.0352±2.3145) μg/mL, respectively, the difference between the two groups was distinguished (P<0.05). Meanwhile, ethosomal carrier was not toxic to the cultured cells. Conclusion Ethosomes can effectively enhance intracellular delivery of 5-FU, as a result can enhance the inhibitory effect of 5-FU on fibroblast viability. Meanwhile, ethosomal carrier was not toxic to the cultured cells. Ethosomes as percutaneous drug delivery carriers, are also a promising candidate for the delivery of biological and chemical compounds to cultured cells.

Key words: Ethosomes, 5-fluorouracil, Hypertrophic scar, Fibroblast, Intracellular delivery

中图分类号: