血管紧张素Ⅱ对大鼠骨髓间充质干细胞向角质细胞分化的调控作用  

Regulatory effects of angiotensin Ⅱ on differentiation of mesenchymal stem cells to keratinocytes

作　　者：吴帆[1] 刘宏伟[1] 程飚[2] 肖丽玲[1] 李升红[1] 廖选[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》2015年第5期1019-1022,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81272100）;广州市科技信息局重大项目（201300000091）

摘　　要：目的 探讨血管紧张素Ⅱ（AngⅡ）对骨髓间充质干细胞（BMSCs）向角质细胞分化的影响及其信号机制.方法 分离培养正常Wistar大鼠BMSCs,流式细胞仪检测BMSCs表面抗原的表达;酶联免疫吸附试验法测定BMSCs中AngⅡ分泌浓度;免疫细胞化学法测定成角质诱导后BMSCs中角蛋白10（K10）的表达.流式细胞仪法检测AngⅡ、AngⅡ1型受体（AT1）阻滞剂Losartan、AngⅡ2型受体（AT2）阻滞剂PD123319及其下游信号分子阻滞剂对BMSCs成角质诱导后K10阳性细胞数的影响.结果 流式细胞仪检测BMSCs表面标志CD29和CD90阳性表达率均在99％及以上,CD34、CD45阳性表达率均低于2％.AngⅡ在细胞上清液中的浓度随培养时间增加呈递增趋势（P＜0.05）.流式细胞仪检测添加AngⅡ的成角质诱导组中BMSCs的K10阳性百分率在7d和14 d分别为69.02％和82.10％,明显高于对照组（P＜0.05）.而添加Losartan及下游信号分子阻滞剂SB203580、AG490、SP600125后,K10阳性细胞数减少（P＜0.05）.结论 AngⅡ可显著提高BMSCs向角质细胞的转化率,p38丝裂原活化蛋白激酶（p38MAPK）、酪氨酸激酶2/3（JAK2/3）、c-jun氨基端激酶（JNK）信号通路与AngⅡ介导的作用密切相关.AngⅡ促进BMSCs向角质细胞转化可能是其促进创面上皮化的机制之一.Objective To investigate the effect of angiotensin Ⅱ （Ang Ⅱ） on the differentiation of bone marrow mesenchymal stem cells （BMSCs） to keratinocytes and the signal mechanism.Methods The BMSCs from Wistar rats were isolated.The expression of BMSCs surface antigens was detected by flow cytometry.The secretion concentration of Ang Ⅱ was detected by enzyme linked immunosorbent assay （ELISA）.The expression of keratin 10 （K10） was detected by immunocytochemistry after keratinocyte induction.The effects of Ang Ⅱ,Losartan,PD123319 and their downstream signal molecule blockers on the number of K10 positive cells after keratinocyte induction by BMSCs were observed.Results The positive rate of BMSCs surface markers CD29 and CD90 was 99% or above.The positive rate of CD34 and CD45 was less than 2%.The concentration of Ang Ⅱ in cell supematant showed an increasing trend with the increase of incubation time （P 〈 0.05）.The K10 positive rate of BMSCs in keratinocyte induction group given Ang Ⅱ in 7 and 14 days was 69.02% and 82.10% respectively,which was obviously higher than that in control group （P 〈 0.05）.The number of positive K10 cells treated with Losartan,SB203580,AG490,or SP600125 was reduced.Conclusion Ang Ⅱ could significantly increase the conversion rate of BMSCs to keratinocytes.p38 mitogen activated protein kinase （p38MAPK）,janus kinase 2/3 （JAK2/3） and c-Jun N-terminal kinase （JNK） signal pathways were closely related with the role mediated by Ang Ⅱ.Ang Ⅱ promoted BMSCs to convert into keratinocytes,which might be one of the mechanisms of promoting wound epithelialization.

关 键 词：血管紧张素Ⅱ 骨髓间充质干细胞 角质细胞 创面愈合 

分 类 号：R965[医药卫生—药理学]