冷刺激对骨髓间充质干细胞增殖及成骨分化的影响  

Effect of cold exposure on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells via the p38 MAPK pathway

作　　者：聂义珍[1] 闫朝岐[1] 付红梅[1] 赵兴鹃[1] NIE Yi-zhen;YAN Zhao-qi;FU Hong-mei;ZHAO Xing-juan(Physical Examination Center,The Second Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第二医院体检中心,哈尔滨150086

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2018年第6期577-583,共7页Chinese Journal Of Osteoporosis And Bone Mineral Research

摘　　要：目的分析冷刺激对于骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)成骨分化的影响,为研究环境温度对于骨质疏松症的影响提供可信的依据。方法选取C57BL/6小鼠,随机分为常温对照组和冷刺激组。实验结束后取双侧股骨、胫骨,全骨髓贴壁法分离BMMSCs并向成骨细胞定向诱导分化。取C57BL/6小鼠的正常BMMSCs体外进行冷刺激,细胞分成37℃组、37℃+抑制剂组[p38丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)抑制剂SB203580]、18℃组、18℃+抑制剂组。结果冷刺激使BMMSCs呈纺锤状和成纤维细胞样改变和细胞增殖水平下降(P<0. 01)。茜素红染色发现,冷刺激7 d后细胞有聚集趋势;体外实验也观察到冷刺激使细胞聚集且染色加深,加入SB203580后,成骨分化减弱。冷刺激提高了成骨分化标志物Runt相关转录因子2 (Runt-related transcription factor2,Runx2)、骨桥蛋白(osteopontin,OPN)、骨涎蛋白(bone sialoprotein,BSP)和胶原蛋白-1 (collagen-Ⅰ) mRNA表达(P<0. 01),SB203580抑制了这些标志物的表达。冷刺激显著诱导了p38 MAPK的磷酸化(P <0. 01),SB203580可下调冷刺激引起的p-p38 MAPK水平的提高(P<0. 01)。结论冷刺激使BMMSCs增殖能力下降,同时活化p38 MAPK通路促进BMMSCs向成骨细胞转化。Objective To investigate the effects of cold exposure on the osteogenic differentiation of bone marrow mesenchymal stem cells （BMMSCs） and the mechanism involved in vitro , which provide reliable data for the effects of environment on osteoporosis. Methods A total of 36 C57BL/6 mice （6-8 weeks old） were randomly divided into two groups the control group and the cold exposure group. After the experiment, we separated the bilateral femur and tibia. BMMSCs were isolated from the bone marrow cavity and incubated with the osteoblast inducing conditional media. BMMSCs were isolated from the normal C57BL/6 mice. BMMSCs were stimulated by the low temperature in vitro . The isolated BMMSCs were divided into four groups： 37 ℃ stimulation group, 37 ℃ stimulation＋SB203580 group （p38 MAPK inhibitor SB203580）, 18 ℃ stimulation group, 18 ℃ stimulation＋SB203580 group. Results BMMSCs had a spindle-shaped and fibroblastlike morphology and low proliferation level （ P 〈0.01）. Alizarin red staining found that the cells under cold exposure had gathered trend and dyed deep. However, this change in the cell was greatly inhibited by p38 MAPK inhibitor SB203580. Furthermore, cold exposure significantly elevated runt-related transcription factor 2（Runx2）, bone sialoprotein（BSP）, osteopontin（OPN） and collagen Ⅰ levels and promoted the phosphorylation of p38 MAPK. However, the inducing effects were greatly inhibited by p38 MAPK inhibitor SB203580. Conclusion Cold exposure inhibits the cell proliferate and promotes the osteogenic differentiation of BMMSCs partially via the p38 MAPK pathway.

关 键 词：冷刺激 骨髓间充质干细胞 成骨分化 

分 类 号：R594.2[医药卫生—内科学]