转化生长因子β1受体抑制剂SD-208对人增生性瘢痕的作用  

Effects of transforming growth factor β1 receptor inhibitor SD-208 on human hypertrophic scar

作　　者：李明[1] 方勇[1] 姚敏[1] 俞为荣[1] 倪涛[1] 顾钏[1] 杨鹏高[1] 毛志刚[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,201999

出　　处：《中华烧伤杂志》2016年第7期389-395,共7页Chinese Journal of Burns

基　　金：国家自然科学基金（81272081）

摘　　要：目的探讨TGF—β1受体抑制剂SD-208对人增生性瘢痕的作用及其机制。方法取。i废弃人增生性瘢痕组织，分离瘢痕Fb，并进行传代培养，采用第5代细胞进行以下实验。（1）按照随机数字表法（分组方法下同）将细胞分为空白对照组及0．5、1．0、3．0、5．0μmol／LSD-208组，每组6孔。空白对照组加入lμLPBS，后4组分别加入1μL0．5、1．0、3．0、5．0μmol／LSD-208，培养12h，细胞计数试剂盒8及酶标仪检测细胞增殖活性（以吸光度值表示）。根据细胞增殖活性结果选取适宜物质的量浓度的SD-208用于后续实验。（2）另取细胞，分为空白对照组及1、3μmol／LSD-208组，同（1）处理，每组8孔，Transwell法检测迁移细胞数。（3）另取细胞，同（2）分组处理，罗丹明-鬼笔环肽染色，观察细胞微丝形态。（4）另取细胞，同（2）分组处理，蛋白质印迹法检测TGF—p，蛋白表达。（5）取48只BALB／c裸鼠，分为生理盐水组24只与μmol／LSD-208组24只，于背部做长度为1cm的纵行切口，埋置人增生性瘢痕组织块。伤后7d，生理盐水组与1μmol／LSD-208组裸鼠瘢痕内分别多点注射0．05mL生理盐水及1Ixmol／LSD-208，每日注射1次。于首次给药后0（当日）、2、4、6、8、10、12、14、16、18、20d，取8只裸鼠，电子秤称量体质量；用游标卡尺测量瘢痕面积，计算剩余瘢痕面积百分比。于首次给药后1、2、3周，取出人增生性瘢痕组织块，免疫组织化学染色法检测TGF-β1。蛋白表达。对数据行单因素方差分析、两独立样本t检验。结果（1）空白对照组及0．5、1．0、3．0、5．0μmol／LSD-208组细胞增殖活性分别为1．00±0．03、0．90±0．08、0．68±0．11、0．54±0．04、0．42±0．09，0．5、1．0、3．0、5．0μmol／LSD-208组细胞增殖活性均明显低于空白对照组（t值为2．9～22．1，P〈0．05或P〈0．01）。（2）1、3μmol�Objective To investigate the effects of transforming growth factor β1 （TGF-β1 ） receptor inhibitor SD-208 on human hypertrophic scar and its mechanisms. Methods Scar fibroblasts were isolated from deprecated human hypertrophic scar tissue and then sub-cultured. Cells of the fifth passage were usedin the following experiments. （ 1 ） Cells were divided into blank control group （BC） and 0.5, 1.0, 3.0, and 5.0 μmol/L SD-208 groups according to the random number table （the same grouping method below） , with 6 wells in each group. Cells in group BC were added with 1 p,L phosphate buffer solution, while cells in the latter four groups were added with 0.5, 1.0, 3.0, and 5.0 μmol/L SD-208, respectively. After being cultured for 12 hours, the proliferation activity of cells was detected by cell counting kit 8 and microplate reader （denoted as absorbance value）. Suitable amount of substance concentration of SD-208 according to the results of proliferation activity of cells was chosen for the following experiments. （2） Another batch of ceils were divided into group BC and 1, 3 μmol/L SD-208 groups and treated as in （ 1 ） , with 8 wells in each group. The number of migration cells was detected by transwell method. （3） Another batch of cells were grouped and treated as in （2） , and the microfilament morphology of cells was observed by rhodamine- phalloidin staining. （4） Another batch of ceils were grouped and treated as in （2）, and the protein expres- sion of TGF-β1 was assessed with Western blotting. （5） Forty-eight BALB/c nude mice were divided into normal saline group （NS） and 1 p, mol/L SD-208 group, and one longitudinal incision with length of 1 cm was made on their back. Then human hypertrophic scar tissue was embedded into the incision. On post inju- ry day 7, muhipoint injection of NS in a volume of 0.05 mL was performed in wounds of rats in group NS, while rats in 1 p, mol/L SD-208 group were given 0.05 mL l μmol/L SD-208, once a day. On the day 0 （the same

关 键 词：瘢痕 转化生长因子Β1 细胞增殖 细胞运动 微丝 SD-208 

分 类 号：R622[医药卫生—整形外科]