中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2014年
12期
1225-1229
,共5页
李芳菲%张波%刘苹%翁土军%罗刚%邓蔓菁
李芳菲%張波%劉蘋%翁土軍%囉剛%鄧蔓菁
리방비%장파%류평%옹토군%라강%산만정
创伤和损伤%间质干细胞%自体受体
創傷和損傷%間質榦細胞%自體受體
창상화손상%간질간세포%자체수체
Wounds and injuries%Mesenchymal stem cells%Autoreceptors
目的 探讨BMSCs膜表面受体CXCR4和CXCR7的表达及其在BMSCs迁移中的作用,为骨创伤修复提供理论依据.方法 选择C57BL/6小鼠经贴壁法培养获得第2代BMSCs,免疫荧光染色法和流式细胞技术检测BMSCs膜表面受体CXCR4和CXCR7的表达.趋化因子CXCL12诱导BMSCs迁移实验分为对照组(无血清培养基诱导BMSCs迁移)、CXCL12组(含CXCL12无血清培养基诱导BMSCs迁移)和CXCR4阻断组(含CXCL12无血清培养基诱导阻断膜受体CXCR4后BMSCs迁移),统计BMSCs迁移数量,观察CXCR4和CXCR7趋化BMSCs迁移的作用.采用Western blot检测阻断膜表面受体CXCR4后CXCR7蛋白的表达.结果 免疫荧光结果显示大量BMSCs膜表面表达CXCR4和CXCR7.流式细胞仪检测结果显示BMSCs膜表面分子CXCR4的阳性率为96.4％,CXCR7的阳性率为97.3％.细胞迁移结果显示,CXCL12组BMSCs迁移量最高,CXCR4阻断组BMSCs迁移量显著低于CXCL12组但迁移量仍高于对照组(P＜0.01),而阻断XCCR4后CXCR7蛋白表达升高.结论 CXCR4和CXCR7受体均表达于BMSCs膜表面,在CXCL12调节BMSCs迁移至骨创伤创面过程中CXCR4发挥主要作用.
目的 探討BMSCs膜錶麵受體CXCR4和CXCR7的錶達及其在BMSCs遷移中的作用,為骨創傷脩複提供理論依據.方法 選擇C57BL/6小鼠經貼壁法培養穫得第2代BMSCs,免疫熒光染色法和流式細胞技術檢測BMSCs膜錶麵受體CXCR4和CXCR7的錶達.趨化因子CXCL12誘導BMSCs遷移實驗分為對照組(無血清培養基誘導BMSCs遷移)、CXCL12組(含CXCL12無血清培養基誘導BMSCs遷移)和CXCR4阻斷組(含CXCL12無血清培養基誘導阻斷膜受體CXCR4後BMSCs遷移),統計BMSCs遷移數量,觀察CXCR4和CXCR7趨化BMSCs遷移的作用.採用Western blot檢測阻斷膜錶麵受體CXCR4後CXCR7蛋白的錶達.結果 免疫熒光結果顯示大量BMSCs膜錶麵錶達CXCR4和CXCR7.流式細胞儀檢測結果顯示BMSCs膜錶麵分子CXCR4的暘性率為96.4％,CXCR7的暘性率為97.3％.細胞遷移結果顯示,CXCL12組BMSCs遷移量最高,CXCR4阻斷組BMSCs遷移量顯著低于CXCL12組但遷移量仍高于對照組(P＜0.01),而阻斷XCCR4後CXCR7蛋白錶達升高.結論 CXCR4和CXCR7受體均錶達于BMSCs膜錶麵,在CXCL12調節BMSCs遷移至骨創傷創麵過程中CXCR4髮揮主要作用.
목적 탐토BMSCs막표면수체CXCR4화CXCR7적표체급기재BMSCs천이중적작용,위골창상수복제공이론의거.방법 선택C57BL/6소서경첩벽법배양획득제2대BMSCs,면역형광염색법화류식세포기술검측BMSCs막표면수체CXCR4화CXCR7적표체.추화인자CXCL12유도BMSCs천이실험분위대조조(무혈청배양기유도BMSCs천이)、CXCL12조(함CXCL12무혈청배양기유도BMSCs천이)화CXCR4조단조(함CXCL12무혈청배양기유도조단막수체CXCR4후BMSCs천이),통계BMSCs천이수량,관찰CXCR4화CXCR7추화BMSCs천이적작용.채용Western blot검측조단막표면수체CXCR4후CXCR7단백적표체.결과 면역형광결과현시대량BMSCs막표면표체CXCR4화CXCR7.류식세포의검측결과현시BMSCs막표면분자CXCR4적양성솔위96.4％,CXCR7적양성솔위97.3％.세포천이결과현시,CXCL12조BMSCs천이량최고,CXCR4조단조BMSCs천이량현저저우CXCL12조단천이량잉고우대조조(P＜0.01),이조단XCCR4후CXCR7단백표체승고.결론 CXCR4화CXCR7수체균표체우BMSCs막표면,재CXCL12조절BMSCs천이지골창상창면과정중CXCR4발휘주요작용.
Objective To investigate the function of CXCR4 and CXCR7 receptors expressed in bone marrow mesenchymal stem cells (BMSCs) membrane surface in process of cell migration,in order to provide a theoretical basis for bone trauma repair.Methods C57BL/6 mice were selected to collect BMSCs of second passage after using the adherence culture method.Expressions of CXCR4 and CXCR7 receptors on BMSCs membrane surface were detected using immunofluorescent staining and flow cytometry.In CXCL12-induced chemotaxis of BMSCs,the assay was divided into control group (cells were seeded in serum-free medium),CXCL12 group and (cells were seeded in serum-free medium containing CXCL12),and CXCR4-blocked group (cells were seeded in serum-free medium containing CXCL12 after the blockade of CXCR4).Migration of BMSCs was qualified and used to determine the chemotaxis role of CXCR4 and CXCR7.After the blockade of CXCR4,expression of CXCR7 was detected using the Western blot method.Results lmmunofluorescence showed overexpressions of CXCR4 and CXCR7 receptors on BMSCs membrane surface.Flow cytometry showed the positive rate of CXCR4 and CXCR7 on BMSCs membrane surface was 96.4％ and 97.3％ respectively.Cell migration assay showed amount of MBSCs migration was the highest in CXCL12 group,relative higher in CXCR4-blocked group and the lowest in control group (P ＜ 0.01).In CXCR4-blocked group,expression of CXCR7 increased.Conclusion CXCR4 and CXCR7 Receptors are expressed in BMSCs membrane surface,but CXCR4 play the major role in CXCL12-induced BMSCs migration to traumatic bone wounds.
