丝裂原活化蛋白激酶通路介导创面渗出液对大鼠表皮干细胞内游离钙浓度的影响  

Mitogen-activated protein kinase is involved in the changes of intracellular free calcium concentration induced by wound exudate in cultured rat epidermal stem cells

作　　者：向启利[1] 李建福[1] 付小兵[2] 

机构地区：[1]第三军医大学西南医院整形科,重庆400038 [2]解放军总医院基础部,北京100853

出　　处：《中国病理生理杂志》2007年第6期1185-1188,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金面上资助项目(No.30570708);国家重点基础研究发展规划资助项目(No.2005CB522603)

摘　　要：目的:探讨创面收集液对大鼠表皮干细胞(ESC)内钙离子的作用以及丝裂原活化蛋白激酶(MAPK)通路对胞内游离钙的影响。方法:以聚氨酯海绵收集成年Wistar大鼠背部全层创面渗出液;以快速黏附分离法体外分离、培养新生Wistar大鼠的表皮干细胞。将培养的表皮干细胞分为5组:①单纯对照组;②单纯创面收集液处理组;③PD98059阻断剂+创面收集液处理组;④SB203580阻断剂+创面收集液处理组;⑤PD98059与SB203580阻断剂+创面收集液处理组。应用特异性Ca2+荧光指示剂Fluo-3/AM负载细胞,激光共聚焦显微镜检测细胞内Ca2+荧光强度,以判断游离钙的浓度。结果:单纯创面收集液处理组ESC的Ca2+荧光强度明显较单纯对照组增加;③、④两组均出现了不同的钙振荡现象;而⑤组则表现为Ca2+荧光强度的迅速降低。结论:创面收集液引起ESC内游离Ca2+浓度增加,MAPK信号通路对ESC胞内钙离子有反馈调节作用。AIM： To observe the effects of harvested wound exudate on intracellular free Ca^＋ in epidermal stem cells （ESCs） in vitro, and to investigate the relationship between mitogen - activated protein kinases （MAPKs） signal pathways and Ca^＋ mobilization in this condition. METHODS： Wound exudate was harvested from the 80 full -thickness wounds produced on both sides of the back in 40 adult Wistar rats. ESCs were isolated, purified from neonatal Wistar rats by referring to the formerly records and binding our ideas. When the cultured cells showed up clone growing, they were divided into five groups as follows： group A： control group （no -treatment） ; group B： only treatment with wound exudate; group C： treatment with wound exudate and PD98059; group D： treatment with wound exudate and SB203580; group E： treatment with wound exudate, PD 98059 and SB203580. Then, the cells were incubated with fluorescence Ca^＋dye fluo - 3/AM at 37 ℃ for 30 min, and measured by using laser scanning confecal microscope. RESULTS ： The results showed that the fluorescent intensity of group B was higher than that in group A. A phenomenon of calcium oscillation was found in group C and group D. Furthermore, a rapid decrease of fluorescent intensity was observed in the cells that were preincubated with PD98059 and SB203580 at the same time. CONCLUSION： Based on above results, we propose that wound exudate can directly induce an increase in intracellular free Ca^＋ concentrations of ESCs. MAPKs signaling pathway has an important function of feedback regulation for free Ca^＋ mobilization of ESCs in this condition, and also is capable of affecting the biological behaviour of epidermal stem cells.

关 键 词：表皮干细胞 CA^+ 有丝分裂素激活蛋白激酶类 

分 类 号：R363[医药卫生—病理学]