干扰素γ缓释微球的制备及对瘢痕疙瘩成纤维细胞增殖和Ⅰ型胶原合成的抑制作用研究  

Preparation of Interferon-γ Sustained-release Microsphere and Inhibitory Effects of It on the Proliferation of Keloid-derived Fibroblasts and the Synthesis of Type Ⅰ Collagen

作　　者：王金[1] 王晓军[1] 

机构地区：[1]北京协和医院整形美容外科,北京100032

出　　处：《中国药房》2013年第37期3510-3512,共3页China Pharmacy

摘　　要：目的:制备负载干扰素γ(INF-γ)的可降解缓释微球,考察其体外缓释性能及对瘢痕疙瘩成纤维细胞(KFs)增殖和胶原合成的抑制作用。方法:采用改良的乳化交联法制备缓释微球,考察其粒径、平均载药量、平均包封率及体外累积释放率。取体外培养24 h的第3代KFs,分为空白对照组(A组,10%胎牛血清+DMEM培养基)、IFN-γ组(B组,含500 u/ml的IFN-γ)、负载相同含量IFN-γ的缓释微球组(C组,含500 u/ml的IFN-γ)及空白微球组(D组),分别进行相应处理后再分别培养5 d,MTT法考察各组KFs细胞增殖抑制率;免疫细胞化学法分析各组KFs细胞Ⅰ型胶原蛋白的平均染色值判断蛋白合成受抑制情况。结果:负载IFN-γ的微球平均粒径为(51.87±1.31)μm,平均载药量为(53.64±3.52)u/mg,平均包封率为(89.72±5.63)%;微球7 d内的累积释放率为91.35%;C组KFs细胞的增殖抑制率(60.75%)明显高于B组(33.88%)(P<0.05);B组KFs细胞Ⅰ型胶原蛋白平均染色值(163.75)较A组(193.75)有统计学意义(P<0.05),C组Ⅰ型胶原蛋白平均染色值(150.25)较A组有统计学意义(P<0.01)。结论:制备的IFN-γ微球缓释性能良好,可持续释放达7 d;该微球可抑制人KFs细胞的增殖,减少其Ⅰ型胶原蛋白的合成,较普通IFN-γ具有更强的抑制能力。OBJECTIVE： To prepare the Interferon-y （1NF-y） degradable sustained-release microsphere, and to investigate its sustained-release performance and inhibitory effect of it on keloid-derived fibroblasts （KFs） and the synthesis of type I collagen. METHODS： The sustained-release microsphere was prepared by modified emulsion cross linking method. Mean particle size, mean drug-loading amount, mean entrapment efficiency and accumulative release rate in vitro were all investigated. Third generation in vitro 24 h cultured KFs were divided into negative control group （group A, 10% fetal bovine serum＋DMEM culture medium）, IFN-γ group （group B, 500 u/ml IFN-γ）, IFN-3,-loading sustained-release microsphere group （group C, including 500 u/ml IFN-γ） and blank microsphere group （group D）. They were given relevant treatment for 5 days. The inhibitory rate of KFs proliferation was measured by MTT method. Immunocytochemical staining method was used to investigate the average staining value of type I collagen protein of KFs for judging the inhibition of the syntnesis of protein. RESULTS： The average diameter of IFN-γ loading microsphere was （51.87 ＋ 1.31） pm with the mean drug-loading amount of （53.64 _＋ 3.52） u/rag. Average entrapment efficiency was （89.72 ＋ 5.63）%. 91.35% of IFN-y was released from the microsphere during 7 days. The inhibitory rate of KFs proliferation （60.75%） in group C was significantly higher than in group B （33.88%） （P〈0.05）. Average staining value of type I collagen protein of KFs （163.75） in group B was significantly different from in group A （193.75） （P〈0.05） ; that of type I collagen pro- tein （150.25） in group C was significantly different from in group A （P〈0.01）. CONCLUSIONS： Prepared IFN-γ microsphere shows good sustained-release performance, and constantly releases for γ days. The microsphere can inhibit the proliferation of KFs, reduce the synthesis of type I collagen protein and has stronger inhibitory ef

关 键 词：干扰素Γ 微球 制备 缓释 瘢痕疙瘩成纤维细胞 增殖 抑制作用 

分 类 号：R944.1[医药卫生—药剂学] R978.7[医药卫生—药学]