摘要
目的比较血小板生成素(TPO)、血小板源性生长因子(PDGF)、白细胞介素-11(IL-11)在促进巨核系增殖、分化及成熟过程中的不同效果,初步探讨巨核细胞分化过程中相关信号通路蛋白的表达。方法用免疫磁珠法分离纯化脐血CD34+细胞,在液体培养体系中经TPO、PDGF-BB、干细胞因子(SCF)I、L-1β、IL-3、IL-6及IL-11等细胞因子组合的诱导,检测有核细胞(NC)总数和CD41+、CD61+表达,蛋白印迹实验(Western blot)分析巨核细胞分化过程中相关信号通路蛋白的表达。结果TPO对巨核系的扩增效果明显强于PDGF和IL-11,而PDGFI、L-11的扩增效果差别不大;含IL-11的因子组,NC及CD41+、CD61+细胞的总数都有明显增加,但CD41+、CD61+细胞比例不高;脐血CD34+细胞在TPO、SCFI、L-1βI、L-3I、L-6及IL-11等因子作用下,随着培养时间的延长,促分裂原活化蛋白激酶42/激酶44(MAPK p42/p44)蛋白表达并无明显改变,蛋白激酶B蛋白亦未见明显变化,但磷酸化的p42和p44的表达却明显增加。结论在体外诱导脐血CD34+细胞向巨核系定向分化过程中,TPO起着最重要的作用;TPO联合其他造血生长因子刺激脐血CD34+细胞向巨核细胞分化和增殖过程中,刺激了MAPK p42/p44分子的磷酸化,从而激活和维持巨核细胞的增殖。
Objective To compare the effects of TPO, PDGF, IL-11 on the proliferation, differentiation and maturation of megakaryocytic （MK） lineage and elucidate the expression of related signal transduction proteins in megakaryocytic differentiation. Methods CD34^＋ cells were isolated from umbilical cord blood by using magnetic activated cell sorting system（MACS）, and expanded with combinations of TPO, PDGF-BB, SCF, IL-1β, IL-3, IL-6 and IL-11 in a liquid culture system. The total number of nucleated cells（NC） and the expression of CD41^＋ , CD61^＋ cells were measured. The expression of related signal transduction proteins were analyzed by Western blot. Results TPO exhibited higher effect on MK lineage than that of PDGF and IL-11. PDGF and IL-11 resulted in a higher total number of NC and CD41^＋ , CD61^＋ cells, but the proportion of CD41^＋ , CD61^＋ cells were low. During culturing with TPO, SCF, IL-1β, IL-3, IL-6 and IL-11, the expression of MAPK p42/p44 and AKT proteins showed no change, but the expression of phosphorylation of p44 and p42 increased significantly. Conclusion Inducing of CD34^＋ cells to MK lineage in vitro, TPO has the most important effect; TPO stimulates the phosphorylation of MAPK p42/p44 proteins, thereby activates and maintains the proliferation of megakaryocytes.
出处
《福建医科大学学报》
2006年第4期330-333,共4页
Journal of Fujian Medical University
基金
福建省自然科学基金资助项目(C0210026)
