MAPK信号通路在淫羊藿甙促进成骨细胞Cbfa1蛋白表达中的作用(英文)  

Role of MAPK signal pathyway in the regulation of Cbfa1 protein expression in osteoblast by Icariin

作　　者：张秀珍[1] 宋利格[1] 王博[1] 

机构地区：[1]同济大学附属同济医院内分泌科

出　　处：《中国骨质疏松杂志》2009年第2期115-122,共8页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金资助项目(30070357)

摘　　要：目的观察淫羊藿甙对体外培养大鼠成骨细胞丝裂原活化蛋白激酶（mitogen—activated proteinkinase，MAPK）信号通路的影响以及MAPK信号通路在淫羊藿甙促成骨细胞核心结合因子α1（corebinding factor-1，Cbfal）蛋白表达中的作用，以探讨淫羊藿甙对成骨细胞作用的信号传导机制。方法用酶消化法分离24h内新生sD大鼠颅盖骨成骨细胞，进行原代培养。在培养液中加入淫羊藿甙（10ng／mL），作用于成骨细胞5min、10min、30min、60min，抽提总蛋白，用Westernblot法检测细胞中ERK、p-ERK、P38和p-P38蛋白的表达。用雌二醇（10^-8mol／L）作用于成骨细胞5min、10min、30min，同上法检测细胞中ERK、P．ERK、P38和p-P38蛋白的表达。用淫羊藿甙（10ng／mL）、雌二醇（10^-8mol／L）和u0126或SB203580单独或共同干预成骨细胞24h，抽提核蛋白，用Westernblot法检测Cbfal蛋白的表达。结果①淫羊藿甙作用于成骨细胞，30min时可促进ERK蛋白的磷酸化，并可持续至60min，和空白组比较，差异有显著性（P〈0．05）；淫羊藿甙作用于成骨细胞，5min时即可促进P38蛋白的磷酸化，在30min时达高峰，并可持续至60min，和空白组比较，差异有显著性（P〈0．05）。②雌二醇作用于成骨细胞，在30min时可促进ERK蛋白的磷酸化，和空白组比较，差异有显著性（P〈0．05）；雌二醇作用于成骨细胞，在10min时可促进P38蛋白的磷酸化，并可持续至30min，和空白组比较，差异有显著性（P〈0．05）。③淫羊藿甙和雌二醇均能促进成骨细胞中Cbfal蛋白的表达，和空白组相比，差异有显著性（P〈0．05）。u0126和SB203580可以抑制淫羊藿甙和雌二醇促进Cbfal蛋白表达的作用。结论①淫羊藿甙和雌二醇均可以激活成骨细胞中ERK／MAPK和P38／MAPK信号通路；②淫羊藿甙和雌二醇均能促进成骨细胞中核转录因子Cbfal的表达，并且ERK／MAPK和P38／MAPKObjective To investigate the effect of icariin （ICA） on mitogen-activated protein kinase （MAPK） signal pathway in rat osteoblasts cultured in vitro and the role of MAPK signal pathway in the icariin-promoting expression of core binding factor-1 （Cbfal） in osteoblasts, and to elucidate the signal mechanism of icariin on osteoblasts. Methods Calvarial osteoblasts were obtained from newborn （ 〈 24 h） SD rats by trypsin-collagenase digestion method. After 5 rain, 10 min, 30 rain, 60 min of treatment with icariin （10 ng/mL） or estrodial （E2） （10^-8 mol/L）, total protein was isolated from osteoblasts and proteins of ERK, p-ERK, P38 and p-P38 were detected by western-blot analysis. Then calvarial osteoblasts were cultured in the medium containing icariin （10 ng/ mL）, estrodial （ 10^-8 mol/L） with or without u0126, SB203580 for 24 h respectively, nucleus protein was isolated from osteoblasts and protein of Cbfal was detected by western-blot analysis. Results 1. The protein of p-ERK in calvarial osteoblasts increased at 30 min and lasted for 60 min （P 〈 0.05, contrast to the blank group） when treating osteoblasts with icariin（ 10 ng/mL） ; the protein of p-P38 in calvarial osteoblasts increased at 5 min and was at the peak at 30 min, and lasted for 60 rain （ P 〈 0.05, contrast to the blank group） when treating osteoblasts with icariin（ 10 ng/mL）. 2. The protein of p-ERK in calvarial osteoblasts increased at 30 min （ P 〈 0.05, contrast to the blank group） when treating osteoblasts with estrodial （ 10^-8 tool/L） ; the protein of p-P38 in calvarial osteoblasts increased at 10 min and continuously increased to 30 min（ P 〈 0.05, contrast to the blank group） when treating osteoblasts with estrodial （10^-8 mol/L）. 3. Both icariin and estrodial could promote the expression of Cbfal protein （P 〈 0.05, contrast to the blank group） and this effect could be weakened by SB203580 or u0126. Conclusion Icariin and estrodial can activate ERK/MAPK and P38/MA

关 键 词：丝裂原活化蛋白激酶 核心结合因子Α1 淫羊藿甙 成骨细胞 

分 类 号：R285.5[医药卫生—中药学]