CAJ | 학술논문

目的探讨preptin对人成骨细胞增殖和分化的影响及其信号途径.方法体外培养人成骨细胞,用10-10、10-9、10-8和10-7mol/L preptin干预24 h,以[3H]脱氧胸腺嘧啶苷掺入法分析细胞增殖,用分光光度计法测定细胞碱性磷酸酶(ALP)活性判断细胞分化程度.Western印迹法检测细胞外信号调节激酶(ERK)、p38丝裂原活化蛋白激酶(p38MAPK)和c-Jun氨基末端激酶(JNK)的磷酸化水平.并在preptin干预前以ERK抑制剂(PD98059)、p38 MAPK抑制剂(SB203580)和JNK抑制剂(SP600125)预处理,观察preptin诱导人成骨细胞增殖和分化的途径.结果 Preptin剂量依赖地增加人成骨细胞的增殖和ALP活性,10-9mol/L浓度时达最大效应(均P<0.01).Preptin刺激人成骨细胞ERK的磷酸化,对p38MAPK和JNK无作用.PD98059阻断preptin刺激的成骨细胞增殖及ALP活性增加(均P<0.05),而SP600125和SB203580无此效应.结论 Preptin通过ERK途径促进人成骨细胞的增殖和分化.
목적 탐토preptin대인성골세포증식화분화적영향급기신호도경.방법 체외배양인성골세포,용10-10、10-9、10-8화10-7mol/L preptin간예24 h,이[3H]탈양흉선밀정감참입법분석세포증식,용분광광도계법측정세포감성린산매(ALP)활성판단세포분화정도.Western인적법검측세포외신호조절격매(ERK)、p38사렬원활화단백격매(p38MAPK)화c-Jun안기말단격매(JNK)적린산화수평.병재preptin간예전이ERK억제제(PD98059)、p38 MAPK억제제(SB203580)화JNK억제제(SP600125)예처리,관찰preptin유도인성골세포증식화분화적도경.결과 Preptin제량의뢰지증가인성골세포적증식화ALP활성,10-9mol/L농도시체최대효응(균P<0.01).Preptin자격인성골세포ERK적린산화,대p38MAPK화JNK무작용.PD98059조단preptin자격적성골세포증식급ALP활성증가(균P<0.05),이SP600125화SB203580무차효응.결론 Preptin통과ERK도경촉진인성골세포적증식화분화.
Objective To investigate the effect of preptin on proliferation and differentiation of human osteoblasts. Methods After human osteoblasts were incubated with 10-10, 10-9, 10-8 , 10-7 mol/L preptin for 24 h,the proliferation of osteoblasts was determined by[3H]thymidine incorporation and alkaline phosphatase (ALP)activity was assayed by spectrophotometric measurement. The phosphorylation levels of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase ( MAPK), extracellular signal-regulated kinase (ERK) 1/2 were assayed by Western blot. ERK inhibitor PD98059, p38MAPK inhibitor SB203580, and JNK inhibitor SP600125were used for investigating the signal pathway of preptin-stimulated osteoblast proliferation and differentiation.Results Preptin dose-dependently increased human proliferation of osteoblasts and ALP activity with the maximum effect at the concentration of l0-9 mol/L (both P<0.01 ). Preptin stimulated ERK phosphorylation in human osteoblasts, but not p38 MAPK and JNK phosphorylation. PD98059 blocked preptin-sitmulated human osteoblasts proliferation and ALP activity (both P<0.05 ), while SB203580 and SP600125 had no effect. Conclusions Preptin promotes the proliferation and differentiation of human osteoblasts through ERK pathway.