黄芪对人皮肤表皮干细胞增殖活性的影响  

Radix Astragalis effects on the proliferative activities of epidermal stem cells derived from human skin

作　　者：胡翔[1] 邹萍[1] 刘莉玲[2] 刘德伍[3] 

机构地区：[1]南昌大学第一附属医院药剂科,江西省南昌市330006 [2]江西省卫生学校,江西省南昌市330029 [3]南昌大学第一附属医院烧伤中心,江西省南昌市330006

出　　处：《中国组织工程研究与临床康复》2009年第19期3689-3692,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30560058);江西省科技厅重大科技招标项目(200604)~~

摘　　要：背景:利用干细胞技术修复创面是当前医学领域治疗伤口愈合的重要策略,黄芪具有扶正固本、益气活血、延缓衰老等作用,被广泛应用于临床各科多种病证的治疗。目的:探讨黄芪对人皮肤表皮干细胞增殖活性的调控作用。设计、时间及地点:细胞分子生物学实验,于2005-09/2008-06在南昌大学第一附属医院完成。材料:烧伤整形患者植皮剩余皮片来源于南昌大学第一附属医院烧伤中心。黄芪注射液为正大青春宝药业有限公司产品,批号0207114,每毫升含黄芪甲苷≥1.0mg,相当于含生药2g。方法:采用胰蛋白酶和乙二胺四乙酸联合消化法分离表皮,Ⅳ型胶原快速贴附法纯化培养人表皮干细胞。实验组以含黄芪、表皮生长因子和角质细胞无血清培养基组成的表皮干细胞培养液进行体外培养,对照组不含黄芪,其余培养条件相同。主要观察指标:计算细胞克隆形成率,流式细胞仪测定细胞周期,免疫细胞化学染色法和图像定量分析法观察端粒酶反转录酶表达的变化。结果:表皮干细胞呈贴壁生长,10d左右有大的细胞克隆形成,实验组人表皮干细胞克隆形成率、G0/G1期细胞比例均明显高于对照组(t=3.17,P<0.01;t=2.83,P<0.05)。两组细胞端粒酶反转录酶均呈阳性表达,定量分析结果显示实验组平均吸光度值和阳性面积值均明显高于对照组(t=2.25,P<0.05;t=4.12,P<0.01)。结论:黄芪可有效增强人皮肤表皮干细胞端粒酶反转录酶的表达和体外增殖活性。BACKGROUND： Repairing wound by using stem cells technology is an important strategy in the wound healing treatment of current medical field. Radix Astragalis, which strengthening body resistance, benefiting vital energy, promoting blood flow and anti-aging, has been widely applied in the treatment of many clinical diseases and syndromes. OBJECTIVE： To investigate the regulatory effect of Radix Astragalis on the proliferation of human epidermal stem cells derived from human skin. DESIGN, TIME AND SETTING： The cell, molecular biology experiment was performed at the First Affiliated Hospital, Nanchang University from September 2005 to June 2008. MATERIALS： Remaining free skin graft of burn and plastic patients was obtained from Burn Center, First Affiliated Hospital, Nanchang University. Radix Astragalis injection solution was purchased from Zhengda Qingchunbao, China （lot number 0207114）, astragaloside ≥1.0mg per milliliter, equal to 2 g crude drug. METHODS： Epidermis was dissociated by Trypsin-ethylenediamine tetraacetic acid （EDTA）. The type Ⅳ collagen was used to isolate and purify the human epidermal stem cells. The cultured cells in experimental group were treated with keratinocyte serum-free medium （KSFM） supplemented with Radix Astragalis and epidermal growth factor in vfro. The procedures were identical in control group, but no Radix Astragalis were added. MAIN OUTCOME MEASURES： The colony forming efficiency was calculated. The cell cycle was determined by the flow cytometer. The expression of the telomerase reverse transcriptase in epidermal stem cells was detected by the immunocytochemistry staining and the image quantitative analysis. RESULTS： Epidermal stem cells presented adhered to the wall, and large cell clone formed at 10 days. The colony forming efficiency of the epidermal stem cells in experimental group and the proportion of the epidermal stem cell in Go/G1 period were significantly higher than in control group （t=3.17, P 〈 0.01; t =2.83, P 〈 0.05）. The telo

关 键 词：表皮干细胞 黄芪 增殖 创面修复 

分 类 号：R394.2[医药卫生—医学遗传学]