解放军医学杂志2018，Vol.43Issue(2)：91-95,5.DOI:10.11855/j.issn.0577-7402.2018.02.01
CREB和ERK1/2在CaMK Ⅱδ调控破骨细胞分化中的作用
Regulation of osteoclast differentiation by CaMK Ⅱδ mediated through CREB and ERK1/2 signal pathways
摘要
Abstract
Objective To investigate the function of cAMP response element binding protein (CREB) and extracellular signal-regulated kinase (ERK1/2) in osteoclast differentiation mediated by Ca2+/calmodulin-dependent kinase Ⅱ (CaMK Ⅱ)δ,and elucidate the molecular mechanism thereof.Methods CaMK Ⅱδ RNA interference lentivirus vector was constructed and mouse RAW264.7 cells were transfected with the virus to determine the interference efficiency.After virus transfection,RAW264.7 cells were treated with 50ng/ml receptor activator of nuclear factor κB ligand (RANKL) and the phosphorylation levels of CREB and ERK1/2 were detected at different time points.The cells were also treated with PD98059,an ERK1/2 inhibitor,to determine the effect of ERK1/2 signal blocking on the expression of nuclear factor activated T-cells cytoplasmic 1 (NFATc1) and osteoclast differentiation.Results Interference efficiency of recombinant CaMK Ⅱδ virus vector was 77.2％ at mRNA level and 70.2％ at protein level.CaMK Ⅱδ RNA interference significantly suppressed phosphorylation of CREB and ERK1/2,and the levels ofp-CREB and p-ERK1/2 were down-regulated by 21％-55％ and 55％-64％,respectively.ERK1/2 inhibitor significantly down-regulated the protein expression of NFATc1,and the number of osteoclast,the number and size of bone resorption lacunae decreased by 39.3％,50.0％ and 52.3％,respectively.Conclusion CaMK Ⅱδ RNA interference may significantly suppress the phosphorylation of CREB and ERK1/2,and CREB and ERK1/2 have mediated the CaMK Ⅱδ-induced osteoclast differentiation.关键词
破骨细胞/钙调蛋白依赖性激酶Ⅱδ/RNA干扰/cAMP反应原件结合蛋白/细胞外信号调节激酶1/2/活化T细胞核因子c1Key words
osteoclast/Ca2+/calmodulin-dependent kinase Ⅱδ/RNA interference/cAMP response element binding protein/extracellular signal-regulated kinase 1/2/nuclear factor of activated T-cells cytoplasmic 1分类
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王红美,董伟,戚孟春,冯晓洁,陆大壮,李任,孙红..CREB和ERK1/2在CaMK Ⅱδ调控破骨细胞分化中的作用[J].解放军医学杂志,2018,43(2):91-95,5.基金项目
国家自然科学基金(81270965) （81270965）
河北省教育厅重点研究项目(ZD2015005) （ZD2015005）
河北省自然科学基金(H2017209114)This work was supported by the National Natural Science Foundation (81270965),the Key Research Project of Hebei Provincial Education Department (ZD2015005),and the Natural Science Foundation of Hebei Province (H2017209114) （H2017209114）
