上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2017, Vol. 37 ›› Issue (7): 924-.doi: 10.3969/j.issn.1674-8115.2017.07.007

• 论著（基础研究） • 上一篇    下一篇

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

胡艳阁，郝健，张迪，杨川峰，宋晨璐，方勇   

  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
  • 出版日期:2017-07-28 发布日期:2017-08-25
  • 通讯作者: 方勇，电子信箱：fangyong1020@hotmail.com
  • 作者简介:胡艳阁（1989—），女，硕士生；电子信箱：huyange89@sina.com
  • 基金资助:
    国家自然科学基金（81272081）

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

HU Yan-ge, HAO Jian, ZHANG Di, YANG Chuan-feng, SONG Chen-lu, FANG Yong   

  1. Department of Burn and Plastic Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China
  • Online:2017-07-28 Published:2017-08-25
  • Supported by:
    National Natural Science Foundation of China，81272081

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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、12.5 μmol/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（COL Ⅰ）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 0.5 μmol/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 COL Ⅰ / 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 0.5 μmol/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 COL Ⅰ的分泌及 TGF-β1 的产生（均 P<0.01）。动物实验表明：① JQ-1 组 COL Ⅰ / Ⅲ总含量较 DMSO 组显著下降，且 COL Ⅰ / 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、COL Ⅰ的分泌及 TGF-β1 的产生；其对裸鼠体内的人瘢痕组织中 COL Ⅰ / 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。

关键词: 增生性瘢痕, 瘢痕成纤维细胞, 溴结构域蛋白 4, JQ-1

Abstract:

 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.

Key words: hypertrophic scar, scar fibroblast, bromodomain containing protein 4, JQ-1