摘要
目的观察力学刺激对MG-63成骨样细胞护骨素(osteoprotegrin,OPG)表达及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路的影响,以及探讨MAPK信号通路在OPG表达中的作用。方法将不同作用时间的机械应变分别作用于MG-63成骨样细胞,采用免疫印迹(Western blot)法检测OPG蛋白、磷酸化的细胞外信号调节激酶1/2 (p-ERK1/2)、p38MAPK (p-p38MAPK)、JNK (p-JNK)表达,采用RT-PCR方法检测OPG mRNA的表达,同时应用信号通路阻断剂观察MAPK各信号通路在OPG表达中的作用。结果机械力学刺激明显促进了MG-63成骨样细胞OPG蛋白及mRNA的表达,加力1、3、6、12 h后OPG蛋白表达分别是对照组的2. 3、4. 2、5. 9、6. 6倍(P<0. 05),加力3、6、12 h后OPG mRNA表达分别是对照组的4. 8、6. 8、7. 5倍(P<0. 05),加力12 h组作用最明显。力学刺激能够激活ERK1/2信号通路,加力60 min后激活最显著,p-ERK1/2表达是对照组的3. 2倍(P <0. 05)。力学刺激也能激活JNK信号通路,加力10 min后开始活化,加力30 min后最显著,加力10、15、30 min后p-JNK表达分别是对照组的2. 9、4. 3、5. 1倍(P<0. 05)。但力学刺激不能激活p38MAPK信号通路。分别应用ERK1/2、JNK的信号通路阻断剂后,发现当抑制ERK1/2信号通路时,OPG蛋白表达也受到了抑制,差异有统计学意义(P<0. 05)。结论机械力学刺激上调了MG-63成骨样细胞OPG蛋白及mRNA的表达,ERK1/2信号通路可能参与了此过程。
Objective To observe the effects of mechanical stimulation on the expression of osteoprotegrin(OPG),MAPK signaling pathway and explore the roles of MAPK signaling pathway in mechanical stimulation-induced OPG expression.Methods Cultured MG-63 osteoblast-like cells were strained for different duration by the four-point bending loader.The protein levels of OPG,phosphorylated ERK1/2,p38MAPK and JNK were detected using Western Blot analysis.The mRNA level of OPG was detected using RT-PCR.We detected the expression of OPG after using the specific inhibitors of ERK1/2 and JNK.Results OPG mRNA and protein levels were significantly increased by mechanical stimulation in MG-63 osteoblasts-like cells.Mechanical stimulation activated ERK1/2 and JNK pathways,but not p38-MAPK in MG63 cells.While inhibition of JNK did not affect cyclic stretch-induced OPG expression,inhibition of ERK1/2 suppressed stretch-induced OPG expression in MG63 cells.Conclusion Mechanical stimulation increases the protein and mRNA expressions of OPG in MG-63 osteoblast-like cells.Inhibiton of ERK1/2 pathway suppresses the OPG expression,suggesting that ERK1/2 pathway plays a role in this process.
作者
杨敏
黄凌云
吕泽平
陈晓
肖立伟
YANG Min;HUANG Ling-yun;LYU Ze-ping;CHEN Xiao;XIAO Li-wei(Department of Internal Medicine,Rehabilitation Hospital Affiliated to National Research Center for Rehabilitation Technical Aids,Beijing 100176,China;Department of Endocrinology,the People's Hospital of Hunan Province,Changsha 410016,China;Center of Stomatology,The Second Xiangya Hospital of Central South University,Changsha 410011,China)
出处
《中华骨质疏松和骨矿盐疾病杂志》
CSCD
北大核心
2019年第1期58-64,共7页
Chinese Journal Of Osteoporosis And Bone Mineral Research
关键词
力学刺激
成骨细胞
护骨素
MAPK信号通路
mechanical stimulation
osteoblast
osteoprotegrin
mitogen-activated protein kinase signaling pathways
