摘要
本研究探讨丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路对间充质干细胞(mesen-chymal stem cell,MSC)向成骨分化的影响。采用骨片法分离培养C57/BL小鼠骨实质来源的MSC;取第4代MSC进行成骨诱导,利用Westernblot检测在MSC向成骨分化过程中MAPK所包含的细胞外信号调节激酶(ex-tracellul arsignal-regulated kinase,ERK)、c-JunN端激酶(c-Jun N-terminal kinase,JNK)和p38通路磷酸化水平的变化,以及分别加入3种相应通路抑制剂PD98059、JNKII和SB203580后碱性磷酸酶(ALP)和钙沉积的相应变化。结果表明:MSC在向成骨分化过程中MAPK通路所包括的ERK、JNK和p38通路均发生活化;加入PD98059后,ALP的表达在诱导早期显著提高,而后无显著改变;加入JNKII后不影响ALP和钙的沉积;而加入SB203580后,可显著抑制MSC中ALP的表达和钙的沉积。结论:p38通路在MSC向成骨分化中起正调控作用,ERK通路可能在早期起负调控作用,而JNK通路在其中未见显著作用。
This study was purposed to investigate the effect of mitogen-activated protein kinase （MAPK） pathway on the osteoblast differentiation of mouse mesenchymal stem cells （MSCs）, MSCs were isolated from mouse compact bone and serially passaged. After being cultured in osteogenic induction medium, the phosphorylation levels of extraceUular signal-regulated kinase （ERK）, c-Jun N-terminal kinase （JNK） and p38 were detected by Western blot. The effects of corresponding pathway inhibitors including PD98059, JNK II and SB203580 on alkaline phosphatase （ALP） and calcium accumulation in the osteoblastic differentiation of MSCs were determined by ALP staining and von kossa staining respectively. The results showed that MAPK pathway including ERK, JNK and p38 was activated in differentiation of MSCs into osteoblasts. ALP activity of MSCs increased in the early phase by addition of PD98059 treatment, whereas ALP activity and calcium accumulation were not observed via JNK II treatment. However, SB203580 strongly inhibited the ALP expression and the calcium accumulation. It is concluded that p38 plays a positive role in the osteogenic differentiation of MSCs, and ERK is probably a negative factor at the early phase of differentiation, but the effect of JNK is not essential.
出处
《中国实验血液学杂志》
CAS
CSCD
2010年第4期981-985,共5页
Journal of Experimental Hematology
基金
973国家重点基础研究资助项目(编号2005CB522705)
863国家高新技术研究发展项目(编号2007AA021109)
国家自然科学基金项目(编号30900568
30600309
30730043)
