补骨脂素对大鼠骨髓间充质干细胞成骨及成脂分化的影响  

Effects of Psoralen on the Osteogenesis and Adipocyte Differentiation of Bone Mesenchymal Stem Cells(BM-MSCs) from Rats

作　　者：张洪跃 周潘宇[1] 汪洋[1] 许硕贵[1] 王秀会 

机构地区：[1]第二军医大学附属长海医院急诊科,上海200433 [2]上海浦东新区周浦医院,上海201318

出　　处：《现代生物医学进展》2017年第15期2813-2816,2832,共5页Progress in Modern Biomedicine

基　　金：上海市卫生局科研计划项目(20134394)

摘　　要：目的:探讨补骨脂素对大鼠骨髓间充质干细胞成骨及成脂分化的影响及其可能机制。方法:选取3月龄无特定病原体级健康雌性SD大鼠25只,通过切除卵巢建立绝经大鼠模型。6周后,通过全骨髓贴壁法分离BMSCs并进行原代和传代培养,传至3代后进行成骨和成脂诱导,并按补骨脂素浓度梯度0、5、10、15、20μmol/L进行处理。细胞增殖2周后,通过碱性磷酸酶(ALP)染色实验和油红O染色观察BMSCs成骨和成脂的分化,应用蛋白免疫印迹法测定核心结合蛋白因子RUNX2、骨钙素(OCN)、增强子结合蛋白β(C/EBP-β)、过氧化物酶体增殖物激活受体γ(PPAR-γ)的表达。结果:成骨诱导BMSCs在补骨脂素的作用下ALP染色呈阳性反应,且补骨脂素的浓度为15μmol/L时阳性反应最强。RUNX2、OCN蛋白的表达随着补骨脂素浓度的升高而升高,差异具有统计学意义(P<0.05)。与空白组比较,成脂诱导BMSCs在补骨脂素的作用下油红O染色阳性反应程度出现下降,补骨脂素浓度为20μmol/L时,油红O染色阳性率最低。C/EBP-β、PPAR-γ蛋白的表达均随着补骨脂素浓度的升高而降低,差异具有统计学意义(P<0.05)。结论:补骨脂素体外可增强BMSCs成骨分化作及抑制BMSCs成脂分化,可能与其调节RUNX2、OCN、C/EBP-β和PPAR-γ蛋白的表达有关。Objective：To investigate the effect of Psoralen on the Osteogenesis and adipocyte differentiation of Bone Mesenchymal Stem cells （BMSCs） from Rats.Methods：Twenty-five 3-month-old specific pathogen free female Sprague-Dawley rats were selected and.The model of rat menopause was established by ovariectomization.Six weeks later,the BMSCs from SD rats were isolated and purified by using attachment method,then BMSCs were used for primary and secondary culture,osteogenic and adipogenic induction were carried out after the third generation.The BM-MSCs were processed by psoralen to make the poralen concentration of 0,5,10,15,20 μmol/L.After 2 weeks of cell proliferation,alkaline phosphatase（ALP）staining and the oil Red O staining were used to determine the situation of Osteogenesis and dipocyte differentiation.After 2 weeks＆#39; induction,the expression of the core binding protein 2 （RUNX2）,osteocalcin （OCN）,a peroxisome proliferator activated receptor gamma （PPAR-γ）,enhancer binding protein beta（C/EBP-β）were detected by western blot.Results：After osteogenic induction：the positive expression of ALP of BMSCs in the 5,10,15,20 μmol/L groups were all increased compared to the blank group,with highest in the 15 μmol/L group.The expression of RUNX2 and OCN protein increased with the increase ofpsoralen concentration,the difference was statistically significant （P〈0.05）.After adipocyte induction：the positive expression of 0il Red O staining of BMSCs in the 5,10,15,20 μmol/L groups were all decreased compared to the blank group,with lowest in the 20 μmol/L group.The expression of C/EBP-β and PPAR-γ protein were reduced with the increasing of psoralen concentration,the difference was statistically significant （P〈0.05）.Conclusion：Psoralen could enhance the osteogenic differentiation and inhibit the adipogenesis of BMSCs,which might be related to mediating the expressions ofRUNX2,OCN,C/EBP-β nd PPAR-γ.

关 键 词：补骨脂素 成骨诱导 成脂诱导 骨髓间充质干细胞 

分 类 号：R33[医药卫生—人体生理学] R68[医药卫生—基础医学]