Kawabata Tetsu, Tokuda Haruhiko, Fujita Kazuhiko, Kainuma Shingo, Sakai Go, Matsushima-Nishiwaki Rie, Kozawa Osamu, Otsuka Takanobu
Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan.
Cell Physiol Biochem. 2017;43(3):1025-1036. doi: 10.1159/000481700. Epub 2017 Oct 2.
BACKGROUND/AIMS: Resveratrol is a polyphenol enriched in the skins of grapes and berries, that shows various beneficial effects for human health. In the present study, we investigated the mechanism behind the epidermal growth factor (EGF)-induced migration of osteoblast-like MC3T3-E1 cells, and the effect of resveratrol on this cell migration.
The cell migration was examined using Boyden chamber, and phosphorylation of each kinase was analyzed by Western blotting.
The EGF-induced migration was suppressed by PD98059, an inhibitor of MEK1/2, as well as SB203580, an inhibitor of p38 MAP kinase, SP600125, an inhibitor of SAPK/JNK, and deguelin, an inhibitor of Akt. In contrast, rapamycin, an inhibitor of upstream kinase of p70 S6 kinase, and fasudil, an inhibitor of Rho-kinase, hardly affected the migration. Resveratrol significantly reduced the EGF-induced migration in a dose-dependent manner. SRT1720, an SIRT1 activator, suppressed the migration by EGF. In addition, resveratrol markedly attenuated the EGF-induced phosphorylation of SAPK/JNK and Akt without affecting the phosphorylation of p44/p42 MAP kinase or p38 MAP kinase. The phosphorylation of SAPK/JNK and Akt induced by EGF was down-regulated by SRT1720.
Our results strongly suggest that resveratrol reduces the EGF-stimulated migration of osteoblasts via suppression of SAPK and Akt, and that the inhibitory effect of resveratrol is mediated in part via SIRT1.
背景/目的：白藜芦醇是一种富含于葡萄和浆果果皮中的多酚，对人体健康具有多种有益作用。在本研究中，我们探讨了表皮生长因子（EGF）诱导成骨样MC3T3-E1细胞迁移的机制，以及白藜芦醇对该细胞迁移的影响。
使用Boyden小室检测细胞迁移，并通过蛋白质印迹法分析各激酶的磷酸化情况。
MEK1/2抑制剂PD98059、p38丝裂原活化蛋白激酶（MAPK）抑制剂SB203580、应激激活蛋白激酶/应激活化蛋白激酶（SAPK/JNK）抑制剂SP600125和Akt抑制剂地谷新均抑制了EGF诱导的迁移。相比之下，p70 S6激酶上游激酶抑制剂雷帕霉素和Rho激酶抑制剂法舒地尔对迁移几乎没有影响。白藜芦醇以剂量依赖的方式显著降低了EGF诱导的迁移。SIRT1激活剂SRT1720抑制了EGF诱导的迁移。此外，白藜芦醇显著减弱了EGF诱导的SAPK/JNK和Akt的磷酸化，而不影响p44/p42 MAPK或p38 MAPK的磷酸化。SRT1720下调了EGF诱导的SAPK/JNK和Akt的磷酸化。
我们的结果强烈表明，白藜芦醇通过抑制SAPK和Akt减少了EGF刺激的成骨细胞迁移，并且白藜芦醇的抑制作用部分通过SIRT1介导。
