羊膜负载表皮干细胞促进糖尿病大鼠创面的愈合  

Amniotic membrane loading epidermal stem cells accelerates wound healing in diabetic rats

作　　者：钟清玲[1,2] 刘德伍[2] 刘繁荣[1] 彭燕[2] 于玫[1] 肖鲁良[1] 

机构地区：[1]南昌大学医学院,江西省南昌市330006 [2]南昌大学第一附属医院烧伤研究所,江西省南昌市330006

出　　处：《中国组织工程研究与临床康复》2010年第32期6010-6014,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30560058);江西省教育厅科技项目(GJJ09115);江西省卫生厅科技项目(20092021)~~

摘　　要：背景:表皮干细胞作为皮肤组织的特异性干细胞,具有强大增殖及多向分化潜能,与创面修复紧密相关。近期研究表明糖尿病皮肤创面愈合过程中表皮干细胞数量减少、活性降低是导致其创面难愈的重要原因。目的:观察表皮干细胞在糖尿病大鼠创面愈合中的作用。方法:分离培养及鉴定SD大鼠表皮干细胞,并以BrdU标记。建立糖尿病SD大鼠创面模型,抽签法随机分为3组:表皮干细胞组创面移植羊膜负载BrdU标记的表皮干细胞;羊膜组创面移植羊膜;空白对照组创面未给予干预。观察创面愈合情况、计算创面愈合率,苏木精-伊红及免疫组织化学SP法检测创面愈合组织中BrdU及增殖细胞核抗原表达。用图像分析软件测量阳性细胞积分吸光度平均值。结果与结论:表皮干细胞组治疗后7d创面缩小明显,治疗后14d创面基本愈合,创面愈合率明显高于羊膜组、空白对照组(P<0.01)。表皮干细胞组创面及新生表皮中可见BrdU阳性细胞,而另两组皮肤创面组织中始终未见BrdU阳性细胞。各组创面组织中可见增殖细胞核抗原阳性细胞表达,但表皮干细胞组的阳性细胞积分吸光度平均值与羊膜组、空白对照组比较差异有显著性意义(P<0.01)。结果证实糖尿病大鼠创面愈合过程中表皮干细胞与创缘表皮移行、创面的上皮化有直接关联,可有效促进其创面愈合。BACKGROUND：As the specific stem cells in skin tissues,epidermal stem cells （ESCs） which posses high proliferative potential and multiple-directional potential play a key role in repairing and rebuilding of the skin. Recent studies showed that the decreased amount and the reduced proliferation and differentiation capacity of ESCs may be one of the important mechanisms of difficult wound recovery in diabetes mellitus. OBJECTIVE：To investigate the effects of ESCs on wound healing of diabetic rats. METHODS：ESCs of Sprague Dawley rats were isolated,cultured,identified and labeled with 5-bromo-2＇-deoxyuridine （BrdU）. The wound models of diabetic SD rats were established,and then divided into ESCs,HAM,blank control groups. Human amniotic membrane （HAM） loading labeled BrdU ESCs were implanted to ESCs group. HAM was implanted to HAM group,but blank control group was not intervened. General situation of wound healing was observed,and the healing rate of wound was calculated. Hematoxylin and eosin staining and immunohistochemical staining （SP method） were used to detect BrdU and expression of proliferating cell nuclear antigen in healing tissues. Mean value of integral absorbance of positive cells was measured with image analysis software. RESULTS AND CONCLUSION：At 7 days after treatment,the wounds of ESCs group reduced significantly. At 14 days after treatment,wounds of ESCs group were almost completely healed. Compared with HAM and blank control groups,the wound healing rate of ESCs group was significantly higher （P 0.01）. BrdU-positive cells in the wounds and newborn epidermis of ESCs group were visible,while wound tissue of HAM and blank control groups had no BrdU-positive cells. Proliferating cell nuclear antigen-positive cells in wound tissue of each group could be seen,but mean value of integral absorbance of positive cells showed significantly differences in ESCs group compared to HAM and blank control groups （P 0.01）. The results have confirmed that ESCs have a direct correlation

关 键 词：表皮干细胞 糖尿病 创面愈合 羊膜 干细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]