血管紧张素转化酶抑制剂对表皮干细胞增殖、分化、迁移的影响  

Effect of angiotensin-converting enzyme inhibitor, captoprol on proliferation, differentiation and migration of human epidermal stem cells

作　　者：肖静[1] 刘宏伟[1] 程飚[2] 肖丽玲[1] 徐媛[1] 李升红[1] 

机构地区：[1]暨南大学附属第一医院整形外科再生医学教育部重点实验室,广州510630 [2]广州军区广州总医院整形外科

出　　处：《中华实验外科杂志》2013年第8期1626-1630,共5页Chinese Journal of Experimental Surgery

基　　金：国家重点基础研究发展规划资助项目（2005CB522603）;国家自然科学基金资助项目（30772257、30973127、81272100）;广州市科技计划资助项目（2012J4100044）

摘　　要：目的观察血管紧张素转化酶（ACE）抑制剂卡托普利对表皮干细胞（ESCs）增殖、迁移、分化的影响，探讨ACE在维持ESCs生物学功能中的作用。方法利用差速贴壁法获得10例儿童的ESCs，进行离体培养，检测ACE在ESCs的表达。用XTT法检测不同浓度（1×10^-5、1×10^-6、1×10^-7、1×10^-8mol／L）ACEI卡托普利对ESCs增殖的影响；体外创伤模型观察1×10^-6 mol／L的卡托普利对ESCs6、12、18、24h各时间段的迁移能力；流式细胞仪检测K10的表达观察1×10^-6 moL／L的卡托普利对ESCs分化的影响。结果培养的细胞经β1-整合素和K19免疫荧光双重标记染色，83．55％细胞为双标记阳性细胞，即ESCs。免疫荧光染色和逆转录．聚合酶链反应（RT—PCFt）结果显示ESCs表达ACE。流式细胞仪定量检测培养的细胞ACE阳性率为74．2％。1×10^-6 mol／L的卡托普利可明显抑制ESCs的增殖（P〈0．05），且在第5天达到峰值。1×10^-6 mol／L的卡托普利可明显抑制细胞的迁移能力（P〈0．05）和克隆能力（P〈0．05）。流式细胞仪检测结果表明，1×10^-6 moL／L的卡托普利并不影响K10的表达（P〉0．05）。结论ACE通过影响ESCs的增殖，迁移从而影响皮肤的损伤修复和自我更新。Objective -To observe the effect of angiotensin-converting enzyme （ACE） inhibitor, captoprol on the proliferation, differentiation and migration of human epidermal stem cells （ ESCs）, and explore the role of ACE in maintaining ESCs biological function. Methods Human ESCs were isolated by differential adhesion method from human skin and cultured in vitro, and the expression of ACE, β1-integrin K19 and K10 in human ESCs was examined by using immunostaining and flow cytometry, xTr cell prolifera- tion assay was used to detect the impact of the different concentrations of ACE inhibitor, captopril on the pro- liferation of ESCs, and the scratch assay was done to evaluate the effect of 1 ×10 -6 mol/L captopril on the migration of ESCs. Flow cytometry was used to detect K10 expression, and the effect of 1 × 10-6 mol/L captopril on the differentiation of ESCs was observed. The role of ACE in maintaining the ESCs biology function was explored. Results The cultured human ESCs were identified through flow cytometry after be- ing marked with β1-integrin and K19 antigens. The number of putative ESCs surface markers, β1-integrin and K19 double-labeled cells accounted for 83.55%. Immumofluorescence and reverse transcription-poly- merase chain reaction （RT-PCR） indicated ACE expression on the ESCs, and flow cytometry revealed that the percentage of cells positive for ACE was 74. 2%. Captopril at a dose of 1 ×10 -6 mol/L inhibited the proliferation of ESCs, reaching the peak at fifth day （P 〈 0. 05）. In addition, Captopril at this dose inhibi- ted the migration （ P 〈 0. 05 ） and cloning ability of ESCs （ P 〈 0. 05 ）. Flow cytometry results revealed that Captopril at 1 × 10 -6 mol/L did not affect the expression of K10 in cultured ESCs （P 〉0. 05）. Conclusion This study revealed that ACE may play an important role in skin injury repair and self-renewal through in- fluencing the proliferation and migration of ESCs.

关 键 词：表皮干细胞 血管紧张素转化酶 增殖 迁移 分化 

分 类 号：R972.4[医药卫生—药品]