μ型阿片受体对人表皮干细胞增殖与迁移的影响  

Effects of μ-opoiod receptor on proliferation and migration of human epidermal stem cells in vitro

作　　者：项晓飞[1] 程飚[2] 唐建兵[2] 彭艳[1] 朱江婷[3] 

机构地区：[1]南方医科大学研究生学院, 广州510515 [2]广州军区广州总医院整形外科全军热区创伤救治与组织修复重点实验室 [3]成都医学院第一附属医院皮肤科

出　　处：《中华实验外科杂志》2013年第5期934-936,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：周家自然科学基金资助项目（30871028、81171812）

摘　　要：目的观察μ型阿片受体的活化对体外培养的人表皮干细胞（hESCs）增殖与迁移的影响。方法分离培养原代hESCs，整合素β1、细胞角蛋白19（CK19）染色鉴定。取第2代hESCs分别用含1nmol／L β-内啡肽的K-SFM培养基（A组）、含10nmoL／L纳洛酮及1nmol／L β-内啡肽的K—SFM培养基（B组）以及单纯的K—SFM培养基（C组）进行培养；连续5d，采用噻唑蓝（MTT）法检测hESCs分裂增殖活性。于培养后24h刮擦生长融合成片的hESCs，制备宽度为100μm的体外单层hESCs缺损模型，模型制备后24、48、72、96h，倒置相差显微镜下观察各组表皮创面愈合，并拍照记录，计算模型制备后72h各组创面愈合率。结果原代培养的hESCs贴壁后呈鹅卵石样生长。免疫荧光染色法检测第2代hESCs的整合素B1、CK19均呈阳性表达。分组培养后，M3-SF法测得第2～4天中A组hESCs分裂增殖活性均高于C组，B组低于C组，各组间差异有统计学意义（P〈0．05）。单层细胞缺损模型制备后24h，各组均可见hESCs迁移越过创缘，其中A组越过创缘的细胞数多于C组，B组少于C组；模型制备72h后，由Image—Pro Plus软件分析各组创面愈合率分别为A组（91．80±0．05）％、B组（79．40±0．06）％、C组（86．30±0．75）％，各组间差异均有统计学意义（P〈0．05）。结论μ型阿片受体活化后可促进hESCs分裂增殖和迁移活动，并可能藉此参与皮肤多种代谢调控过程。Objective To investigate the affects of μ-opioid receptor activation on proliferation and migration of epidermal stem cells （ESCs） in vitro, Methods Primary human ESCs （hESCs） were isolated and cultured, and marked by both integrin β1 and eytokeratin 19 （CK19） to be identified. The hESCs of the second generation were cultnred with K-SFM supplemented with 1 nmol/L β-endorphins in group A, with K-SFM supplemented with 10 mnol/L naloxone and 1 nmol/L β-endorphins in group B, and with K-SFM supplemented with no drugs in group C. The proliferative ability of hESCs in each group was determined by using methyl thiazol tetrazolium （MTT） assay for 5 consecutive days. The 100 μm scratch＇ wounds were created by scraping confluent hESCs plated on Petri-dishes with a sterile pipette tip in vitro. The pictures of migrating eells from the wound edge were taken at 24, 48, 72 and 96 h after incubation with different media. The wound healing rate was calculated by Image-Pro Plus software at 72 h. Results Cultured primary hESCs could adhere to the wall and showed cobblestone-like shape. Immunofluorescenee staining showed the positive results for integrin β1 and CK19. The proliferation rate of hESCs detected by using MTF assay was significantly higher in group A than in groups C, visibly higher in group C than in group B during the day 2 to day 5, and the difference between each two groups was statistically significant （ P 〈 0. 05 ）. In group A, the number of cells migrating the wound edge was greater than in group C, and less in group B than in group C at 24 h. The wound healing rate in groups A, B and C at 72 h was （91.80 ±0. 05）%, （79. 40 ±0. 06）%, and （86. 30 ±0.75）% respectively, showing significant differences among groups （P 〈 0. 05）. Conclusion The activation of β-opioid receptor can promote the proliferation and migration of hESCs in vitro. The β-opioid receptor may be involved in some modutory processes of skin metabolisms.

关 键 词：表皮干细胞 阿片受体 增殖 迁移 

分 类 号：R3[医药卫生—基础医学]