Preptin对人成骨细胞增殖和分化的影响及机制研究  

The mechanism of the effect of preptin on proliferation and differentiation of human osteoblasts

作　　者：朱佳花[1] 刘幼硕[1] 袁凌青[2] 詹俊鲲[1] 汪化文[1] 廖二元[2] 

机构地区：[1]中南大学湘雅二医院老年内分泌科,长沙410011 [2]中南大学湘雅二医院内分泌科、中南大学代谢内分泌研究所

出　　处：《中华内分泌代谢杂志》2010年第9期784-787,共4页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金资助项目（30871191,30801174）;湖南省自然科学基金资助项目（09JJ3037）;教育部博士点新教师基金（200805331017）

摘　　要：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.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.

关 键 词：PREPTIN 人成骨细胞 细胞外信号调节激酶 细胞增殖 

分 类 号：R285[医药卫生—中药学]