不同培养条件对人增生性瘢痕成纤维细胞生物学活性的影响  

Effect of different culture conditions on biological characteristics of fibroblasts derived from human hypertrophic scar

作　　者：董亮 张迪[1] 方勇[1] Dong Liang;Zhang Di;Fang Yong(Department of Burns and Plastic Surgery,Shanghai Ninth People＇s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 201999,China)

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：《中华整形外科杂志》2018年第7期558-564,共7页Chinese Journal of Plastic Surgery

摘　　要：目的探讨不同培养条件对人增生性瘢痕成纤维细胞（hypertrophic scar fibroblast，HSF）生物学活性的影响。方法体外常规培养人HSF。取第4-6代细胞按随机数字表法分为10％氧体积分数＋标准培养基（对照组）、7％氧体积分数＋70％浓度的标准培养基（实验组1）、4％氧体积分数＋40％浓度的标准培养基（实验组2）、1％氧体积分数＋10％浓度的标准培养基（实验组3），每组样本数均为4。先将各组成纤维细胞用10％氧体积分数＋标准培养基培养48h后，再采用不同培养条件分别培养。应用噻唑蓝法、流式细胞术和钙依赖磷脂结合蛋白异硫氰酸荧光素／碘化丙啶双染法、羟脯氨酸试剂盒，蛋白质印迹法分别检测各组成纤维细胞的细胞活力、s期细胞比例、活细胞比例、胶原合成和转化生长因子-β，（TGF-β1）的含量。用SPSS13．0软件对数据进行统计学处理，数据采用单因素方差分析，组内多个均数两两比较采用SNK法。结果①对照组、实验组1、实验组2和实验组3细胞活力分别为0．72±0．03、0．79±0．02、0．75±0．02和0．38±0．03，与对照组比较，实验组1细胞活力较高（P〈0．05），实验组2差异无统计学意义（P〉0．05），实验组3细胞活力较低（P〈0．05）。组内整体比较差异有统计学意义（F：163．32，P〈0．01）。②上述4组胶原合成水平分别为0．42±0．01、0．56±0．01、0．48±0．Ol和0．31±0．01，与对照组比较，实验组1和实验组2胶原合成水平含量均较高（P〈0．05），实验组3的胶原合成水平较低（P〈0．05）。组内整体比较差异有统计学意义（F=357．69，P〈0．01）。③上述4组s期比例分别为（19．40±0．37）％、（22．83±0．28）％、（20．94±0．38）％和（14．54±0．21）％，与对照组比较，实验组1S期比例较高（P〈0．05），实验组2s期比例差异无统计学意义（P�Objective To investigate the effect of different culture conditions on cell proliferation, apoptosis, collagen synthesis and TGF-β1 expression of fibroblasts derived from human hypertrophic scar（HSF）. Methods Fibroblasts were isolated from human hypertrophic sear and cultured in vitro. Cells of passage 4 to 6 were cultured with 10% 02 ＋ 100% culture medium for 48 h, and then divided into four groups, 10% 02 ＋ 100% culture medium group （control group） , 7% 02 ＋ 70% culture medium group （ experimental group 1 ） , 4% 02 ＋ 40% culture medium group （ experimental group 2） and 1% 02 ＋ 10% culture medium group （ experimental group 3） according to the different culture conditions. There are 4 samples in each group. MTT assay, Flow cytometer and the Annexin V-FITC/PI double- staining were performed to detect the cell proliferation, S phase cell ratio and cell apoptosis. Hydroxyproline assay kit was adopted to detect the collagen synthesis of the fibroblasts. Protein expression levels of transforming growth factorβ1 （TGF -β1 ）were determined with Western-blotting. SPSS 13. 0 software was used to analyze the data. Analysis of variance （ oneway ANOVA） and SNK test were used to evaluate significant differences among these groups. Results ②Compared with control group, the cell proliferation rate was higher in the experimental group 1 （P 〈 O. 05 ） , while it significantly decreased in experimental group 3 （ P 〈 0. 05 ）. There was a statistically significant difference among groups （ F = 163.32,P 〈0.01）.）The collagen synthesis was much higher in experimental group 1 compared with control group （0.56 ±0.01 vs 0.42 ±0.01 , P 〈0.05） , and was lower in experimental group 3 （0.48 ±O. 01 ,P 〈 0.05 ）. There was a statistically significant difference among groups （ F = 357.69, P 〈 0.01 ）. ③The S phase cell ratio in the four groups were （ 19.40± 0.37 ） % , （22.83 ±0.28 ） % , （20, 94 ± 0.38 ） % and（ 14.54 ±0.21 ） % respectively. Compa

关 键 词：增生性瘢痕 成纤维细胞 缺氧 培养条件 

分 类 号：R622[医药卫生—整形外科]