人增生性瘢痕与正常皮肤中表皮干细胞的生物学特性及含量比较  

Comparison of biological characteristics and quantity of epidermal stem cells from hypertrophic scar skin and normal skin of human beings

作　　者：周树萍 蔡景龙 牛扶幼 宗宪磊 徐静静 都乐 陈光宇 

机构地区：[1]中国医学科学院整形外科医院整形十六科,北京100144

出　　处：《中华医学杂志》2014年第14期1097-1100,共4页National Medical Journal of China

基　　金：北京市自然科学基金（7122138）

摘　　要：目的：比较人增生性瘢痕与正常皮肤中表皮干细胞的生物学特性及含量。方法增生性瘢痕（瘢痕组）和正常皮肤（对照组）各20例，分别提取和分选表皮干细胞，对两组表皮干细胞采用倒置相差显微镜分别观察其形态特点和检测其生长曲线，采用Western印迹法分别检测其角蛋白19、核蛋白p63、整合素β1蛋白表达的吸光度（ A）值，采用荧光实时定量PCR法分别检测其Oct-4基因和Nanog基因的表达，采用流式细胞术分别检测其整合素CD29、CD49f及角蛋白19表达阳性的细胞比例，并比较其差异。结果两组原代培养的表皮干细胞形态和生长能力基本一致，瘢痕组和对照组表皮干细胞中角蛋白19、核蛋白p63、整合素β1蛋白表达的A值分别为：860±4、712±3、422±6和862±3、707±9、413±6（均P＞0.05），Oct-4、Nanog基因的表达量分别为（7.79±0.44）×10-4、（5.96±0.36）×10-4和（7.93±0.29）×10-4、（6.06±0.35）×10-4（均P＞0.05），CD29、CD49f、角蛋白19表达阳性的细胞比例分别为（97.3±0.7）％、（94.6±1.1）％、（92.5±0.8）％和（98.8±4.6）％、（98.9±0.4）％、（94.4±0.7）％（均P＜0.05）。结论增生性瘢痕表皮干细胞的生物学特性与正常皮肤表皮干细胞的生物学特性基本一致，但含量显著减少。Objective To compare the biological characteristics of epidermal stem cells （ ESCs ） from hypertrophic scar and normal skin.Methods Epidermal stem cells were separated , enriched and adhered from 20 hypertrophic scars （ scar group ） and 20 normal skins （ control group ）.The morphology and growth characteristics of primary epidermal stem cells were observed.The absorbance （ A ） values of expression of Keratin 19, nucleoprotein p63 and integrin β1 were tested by Western blot.And the genes of Oct-4 and Nanog were tested by quantitative reverse transcription-polymerase chain reaction （ qRT-PCR ）.Flow cytometry was used to examine the markers of integrin CD 29, integrin CD49f and Keratin 19 of ESCs.Results The primary cultured cells showed the similar shape and growth curve.By comparing the epidermal stem cells from scar groups and control groups , the absorbance values of the expression of Keratin 19, nucleoprotein p63 and integrin β1 were 860 ±4, 712 ±3, 422 ±6 and 862 ±3, 707 ±9, 413 ±6 （ all P〉0.05）.The expression values of Oct-4, Nanog were （7.79 ±0.44） ×10 -4 , （5.96 ±0.36） ×10 -4 and （7.93 ±0.29） ×10 -4, （6.06 ±0.35） ×10 -4 （all P〉0.05）.Percentages of positive cells expressing CD29, CD49f and Keratin 19 were （97.3 ±0.7）%, （94.6 ±1.1）%, （92.5 ±0.8）% and （98.8 ± 4.6）%, （98.9 ±0.4）%, （94.4 ±0.7）% respectively （all P 〈0.05）.Conclusions The ESCs in hypertrophic scar have the same characteristics with ESCs in normal skin.However , the ESCs from hypertrophic scar are lower than that from normal skin.

关 键 词：瘢痕 干细胞 表皮 皮肤 

分 类 号：R363[医药卫生—病理学]