溴结构域蛋白4特异性抑制剂JQ-1对人增生性瘢痕的影响  

Effects of bromodomain containing protein 4 specific inhibitor JQ-1 on human hypertropic scar

作　　者：胡艳阁[1] 郝健[1] 张迪[1] 杨川峰[1] 宋晨璐 方勇[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院烧伤整形科,上海201999

出　　处：《上海交通大学学报（医学版）》2017年第7期924-929,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81272081)~~

摘　　要：目的·探究溴结构域蛋白4(BRD4)特异性抑制剂JQ-1对人增生性瘢痕的影响及其可能机制。方法·收集人皮肤增生性瘢痕标本,原代培养皮肤瘢痕成纤维细胞,并进行传代培养,予以0.1、0.5、1.0、2.0、2.5、12.5μmol/L不同浓度的JQ-1干预48 h,CCK-8试剂盒、细胞划痕试验和ELISA法分别检测JQ-1对成纤维细胞增殖、迁移、Ⅰ型胶原蛋白(COLⅠ)分泌量及TGF-β1的蛋白水平。36只裸鼠构建瘢痕动物模型,将临床上获得的瘢痕组织(1.0 cm×1.0 cm×0.5 cm)移植于裸鼠背部皮下。继续饲养4周后,将裸鼠分成2组,JQ-1组和DMSO组,分别每日予以0.5μmol/L JQ-1及0.1%DMSO的瘢痕内多点注射,1 mL/只。天狼猩红染色和免疫组织化学法分别观察瘢痕中COLⅠ/Ⅲ及α平滑肌肌动蛋白(α-SMA)的变化情况。结果·细胞实验表明:(1)0.5μmol/L及以上浓度JQ-1对成纤维细胞增殖有明显抑制作用(均P<0.01)。(2)JQ-1可抑制成纤维细胞的迁移活性(均P<0.01)。(3)JQ-1可抑制成纤维细胞COLⅠ的分泌及TGF-β1的产生(均P<0.01)。动物实验表明:(1)JQ-1组COLⅠ/Ⅲ总含量较DMSO组显著下降,且COLⅠ/Ⅲ比值下降(均P<0.05)。(2)JQ-1组α-SMA的含量较对照组明显下降(均P<0.05)。结论·JQ-1可抑制体外人瘢痕成纤维细胞的增殖、迁移、COLⅠ的分泌及TGF-β1的产生;其对裸鼠体内的人瘢痕组织中COLⅠ/Ⅲ的分泌及成纤维细胞向肌成纤维细胞的转化亦有抑制作用。Objective · To investigate effect and the possible molecular mechanism of JQ1, a specific inhibitor of bromodomain containing protein 4, on human hypertropic scar. Methods · Primary fibroblasts were isolated from human hypertrophic scars and treated with JQ-1 of different concentrations （0.1, 0.5, 1.0, 2.0, 2.5, and 12.5 μmol/L） for 48 h. Then CCK-8 kit and wound healing assay were used to measure proliferation and migration of the fibroblasts. ELISA was adopted to detect the levels of collagen type Ⅰ （COL Ⅰ） and TGF-β1 after JQ-1 treatment for 24 h. Thirty-six nude mice were used for hypertrophic scar models. Human hypertrophic scars （1.0 cm×1.0 cm×0.5 cm） were grafted subcutaneously at the backs of nude mice to establish scar animal models. After 4 weeks, the nude mice were averagely divided into two groups, i.e. JQ-1 group and DMSO group, which were respectively injected with 0.5 μmol/L JQ-1 and 0.1% DMSO each mouse every day. COL Ⅰ / Ⅲ and α-smooth muscle actin （α-SMA） were examined by immunohistochemical method and sirius red staining. Results · Cell experiments showed that JQ-1 with the concentration of 0.5 μmol/L and above significantly inhibited proliferation of fibroblasts （P〈0.01）. JQ-1 inhibited migration of fibroblast （P〈0.01）. JQ-1 inhibited secretion of COL Ⅰ and TGF-β1 of fibroblasts （P〈0.01）. Animal experiments showed that concentration and proportion of COL Ⅰ / Ⅲ in JQ-1 group decreased compared to DMSO group （P〈0.05）. α-SMA protein expression in JQ-1 group also decreased （P〈0.05）. Conclusion · JQ-1 can inhibit proliferation, migration, secretion of COL Ⅰ , and production of TGF-β1 of human scar fibroblasts in vitro; it can also inhibit secretion of COL Ⅰ / Ⅲ and fibroblast-myofibroblast differentiation in the human hypertrophic scars in nude mice.

关 键 词：增生性瘢痕 瘢痕成纤维细胞 溴结构域蛋白4 JQ-1 

分 类 号：R622[医药卫生—整形外科]