中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
5期
1019-1022
,共4页
吴帆%刘宏伟%程飚%肖丽玲%李升红%廖选
吳帆%劉宏偉%程飚%肖麗玲%李升紅%廖選
오범%류굉위%정표%초려령%리승홍%료선
血管紧张素Ⅱ%骨髓间充质干细胞%角质细胞%创面愈合
血管緊張素Ⅱ%骨髓間充質榦細胞%角質細胞%創麵愈閤
혈관긴장소Ⅱ%골수간충질간세포%각질세포%창면유합
Angiotensin Ⅱ%Mesenchymal stem cells%Keratinocyte%Wound healing
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.
目的 探討血管緊張素Ⅱ(AngⅡ)對骨髓間充質榦細胞(BMSCs)嚮角質細胞分化的影響及其信號機製.方法 分離培養正常Wistar大鼠BMSCs,流式細胞儀檢測BMSCs錶麵抗原的錶達;酶聯免疫吸附試驗法測定BMSCs中AngⅡ分泌濃度;免疫細胞化學法測定成角質誘導後BMSCs中角蛋白10(K10)的錶達.流式細胞儀法檢測AngⅡ、AngⅡ1型受體(AT1)阻滯劑Losartan、AngⅡ2型受體(AT2)阻滯劑PD123319及其下遊信號分子阻滯劑對BMSCs成角質誘導後K10暘性細胞數的影響.結果 流式細胞儀檢測BMSCs錶麵標誌CD29和CD90暘性錶達率均在99％及以上,CD34、CD45暘性錶達率均低于2％.AngⅡ在細胞上清液中的濃度隨培養時間增加呈遞增趨勢(P＜0.05).流式細胞儀檢測添加AngⅡ的成角質誘導組中BMSCs的K10暘性百分率在7d和14 d分彆為69.02％和82.10％,明顯高于對照組(P＜0.05).而添加Losartan及下遊信號分子阻滯劑SB203580、AG490、SP600125後,K10暘性細胞數減少(P＜0.05).結論 AngⅡ可顯著提高BMSCs嚮角質細胞的轉化率,p38絲裂原活化蛋白激酶(p38MAPK)、酪氨痠激酶2/3(JAK2/3)、c-jun氨基耑激酶(JNK)信號通路與AngⅡ介導的作用密切相關.AngⅡ促進BMSCs嚮角質細胞轉化可能是其促進創麵上皮化的機製之一.
목적 탐토혈관긴장소Ⅱ(AngⅡ)대골수간충질간세포(BMSCs)향각질세포분화적영향급기신호궤제.방법 분리배양정상Wistar대서BMSCs,류식세포의검측BMSCs표면항원적표체;매련면역흡부시험법측정BMSCs중AngⅡ분비농도;면역세포화학법측정성각질유도후BMSCs중각단백10(K10)적표체.류식세포의법검측AngⅡ、AngⅡ1형수체(AT1)조체제Losartan、AngⅡ2형수체(AT2)조체제PD123319급기하유신호분자조체제대BMSCs성각질유도후K10양성세포수적영향.결과 류식세포의검측BMSCs표면표지CD29화CD90양성표체솔균재99％급이상,CD34、CD45양성표체솔균저우2％.AngⅡ재세포상청액중적농도수배양시간증가정체증추세(P＜0.05).류식세포의검측첨가AngⅡ적성각질유도조중BMSCs적K10양성백분솔재7d화14 d분별위69.02％화82.10％,명현고우대조조(P＜0.05).이첨가Losartan급하유신호분자조체제SB203580、AG490、SP600125후,K10양성세포수감소(P＜0.05).결론 AngⅡ가현저제고BMSCs향각질세포적전화솔,p38사렬원활화단백격매(p38MAPK)、락안산격매2/3(JAK2/3)、c-jun안기단격매(JNK)신호통로여AngⅡ개도적작용밀절상관.AngⅡ촉진BMSCs향각질세포전화가능시기촉진창면상피화적궤제지일.
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.
