中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2008年
12期
811-814
,共4页
陈龙%程范军%唐俊明%张卫国%刘歧焕%曾琴兵%孔霞%郭凌郧%王家宁
陳龍%程範軍%唐俊明%張衛國%劉歧煥%曾琴兵%孔霞%郭凌鄖%王傢寧
진룡%정범군%당준명%장위국%류기환%증금병%공하%곽릉운%왕가저
间充质干细胞%迁移%蛋白激酶C%信号转导
間充質榦細胞%遷移%蛋白激酶C%信號轉導
간충질간세포%천이%단백격매C%신호전도
Mesenchymal stem cells%Migration%PKC%Signal transduction
目的 利用Transwell体外迁移体系初步探索EPO在骨髓源间充质干细胞(MSC)迁移中的作用及其信号传递机制.方法 采用经典的全骨髓细胞贴壁法培养MSC,通过成骨、成脂肪等多向诱导分化以及流式细胞术分析其表面标志(CD133、CD34、CD90、CD105)等鉴定MSC特征;以第3代MSC为实验材料,利用Transwell体外迁移体系,先观察不同浓度的rhEPO对MSC迁移的影响,随后在50 nmol/L Wortmannin、50μmol/L PD98059、10μmoL/L U73122、4μg/ml抗EPO-R IsG、30 μmol/LSB203580、10 mmol/L Straurosporine、6 nmol/L G0697、50 μmol/L Pseudo Z等不同的信号转导途径阻断剂的干预下观察EPO对迁移的影响.结果 培养的MSC呈现出CD90、CD105强阳性,具有成骨、成脂肪等多向分化能力;MSC体外迁移能力随着rhEPO浓度(1、10、100、1000 IU/ml)的递增而逐渐增强,并且rhEPO浓度在100 U/ml时,MSC迁移到滤膜上的细胞数接近于峰值;Wortmannin、PD98059、抗EPO-R IgG、Straurosporine、G06976、Pseudo Z对MSC迁移均有影响,其中U73122、抗EPO-R IgG、Straurosporine、Pseudo Z对MSC迁移阻断的效应最显著.结论 EPO-EPO-R所介导的MSC迁移与丝裂原活化蛋白激酶、磷脂酰肌醇特异性磷脂酶C和蛋白激酶C-ζ(PKC-ζ)等信号传递途径有关,且PKC-ζ途径可能处于中心环节.
目的 利用Transwell體外遷移體繫初步探索EPO在骨髓源間充質榦細胞(MSC)遷移中的作用及其信號傳遞機製.方法 採用經典的全骨髓細胞貼壁法培養MSC,通過成骨、成脂肪等多嚮誘導分化以及流式細胞術分析其錶麵標誌(CD133、CD34、CD90、CD105)等鑒定MSC特徵;以第3代MSC為實驗材料,利用Transwell體外遷移體繫,先觀察不同濃度的rhEPO對MSC遷移的影響,隨後在50 nmol/L Wortmannin、50μmol/L PD98059、10μmoL/L U73122、4μg/ml抗EPO-R IsG、30 μmol/LSB203580、10 mmol/L Straurosporine、6 nmol/L G0697、50 μmol/L Pseudo Z等不同的信號轉導途徑阻斷劑的榦預下觀察EPO對遷移的影響.結果 培養的MSC呈現齣CD90、CD105彊暘性,具有成骨、成脂肪等多嚮分化能力;MSC體外遷移能力隨著rhEPO濃度(1、10、100、1000 IU/ml)的遞增而逐漸增彊,併且rhEPO濃度在100 U/ml時,MSC遷移到濾膜上的細胞數接近于峰值;Wortmannin、PD98059、抗EPO-R IgG、Straurosporine、G06976、Pseudo Z對MSC遷移均有影響,其中U73122、抗EPO-R IgG、Straurosporine、Pseudo Z對MSC遷移阻斷的效應最顯著.結論 EPO-EPO-R所介導的MSC遷移與絲裂原活化蛋白激酶、燐脂酰肌醇特異性燐脂酶C和蛋白激酶C-ζ(PKC-ζ)等信號傳遞途徑有關,且PKC-ζ途徑可能處于中心環節.
목적 이용Transwell체외천이체계초보탐색EPO재골수원간충질간세포(MSC)천이중적작용급기신호전체궤제.방법 채용경전적전골수세포첩벽법배양MSC,통과성골、성지방등다향유도분화이급류식세포술분석기표면표지(CD133、CD34、CD90、CD105)등감정MSC특정;이제3대MSC위실험재료,이용Transwell체외천이체계,선관찰불동농도적rhEPO대MSC천이적영향,수후재50 nmol/L Wortmannin、50μmol/L PD98059、10μmoL/L U73122、4μg/ml항EPO-R IsG、30 μmol/LSB203580、10 mmol/L Straurosporine、6 nmol/L G0697、50 μmol/L Pseudo Z등불동적신호전도도경조단제적간예하관찰EPO대천이적영향.결과 배양적MSC정현출CD90、CD105강양성,구유성골、성지방등다향분화능력;MSC체외천이능력수착rhEPO농도(1、10、100、1000 IU/ml)적체증이축점증강,병차rhEPO농도재100 U/ml시,MSC천이도려막상적세포수접근우봉치;Wortmannin、PD98059、항EPO-R IgG、Straurosporine、G06976、Pseudo Z대MSC천이균유영향,기중U73122、항EPO-R IgG、Straurosporine、Pseudo Z대MSC천이조단적효응최현저.결론 EPO-EPO-R소개도적MSC천이여사렬원활화단백격매、린지선기순특이성린지매C화단백격매C-ζ(PKC-ζ)등신호전체도경유관,차PKC-ζ도경가능처우중심배절.
Objective To explore the effects of rhEPO on the migration of bone marrow (BM) derived mesenchymal stem cells(MSCs) and its probable signal transduction mechanism.Methods MSC was cultured by classical whole BM adherence method; MSC characteristics was identified by multi-differentiation and surface marker (CD90,CD133,CD34,CD105).The effect of different concentrations EPO (1,10,100,1000 IU/ml) on MSCs migration were observed.Then 30 minutes later,MSC were treated with signal transduction pathway inhibitors,50 nmol/L wortmannin,50 μmol/L PD98059,10 μmol/L U73122,4 μg/ml Anti EPO-R lgG,30 μmol/L SB203580,10 mmol/L Straurosporine,6 nmol/L G06976 and 50 μmol/L Pseudo Z,respectively.The efficacy of MSC migration was analyzed by Transwell in vitro migration assay.Results Cultured MSCs had the capacities for osteogenic and adipogenic differentiation and highly expressed CD105,CD90 and EPO-R.The efficiency of MSC in vitro migration increased gradnally in a concentrationdependent manner with increasing concentration of rhEPO,and the ability peaked at a concentration of 100 IU/ml.Furthermore,the migration ability was decreased on treated with U73122,Anti EPO-R IgG,Straurosporine,Pseudo Z treatment.Conclusion EPO/EPO-R-mediated MSCs migration is related with MAPK,PIPLC/PKC-ζ signal pathways,PKC-ζ signal pathway may be of central role for MSCs migration.
