Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-alpha and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells

doi:10.1016/j.phymed.2019.153146

	Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-alpha and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells
	Wu, Qing-Chang 1,2; Tang, Xi-Yang 1,2; Dai, Zi-Qin 1,2; Dai, Yi 1,2,3; Xiao, Hui-Hui 4; Yao, Xin-Sheng 1,2
刊名	PHYTOMEDICINE
	2020-03-01
卷号	68 页码:12
关键词	Sweroside Differentiation and mineralization Membrane estrogen receptor-alpha GPR30 P38 signaling pathway MC3T3-E1 cells
ISSN号	0944-7113
DOI	10.1016/j.phymed.2019.153146
通讯作者	Dai, Yi(daiyi1004@163.com) ; Xiao, Hui-Hui(huihui.xiao@polyu.edu.hk)
英文摘要	Background: Dipsaci Radix has been clinically used for thousands of years in China for strengthening muscles and bones. Sweroside is the major active iridoid glycoside isolated from Dipsaci Radix. It has been reported that sweroside can promote alkaline phosphatase (ALP) activity in both the human osteosarcoma cell line MG-63 and rat osteoblasts. However, the underlying mechanism involved in these osteoblastic processes is poorly understood. Purpose: This study aimed to characterize the bone protective effects of sweroside and to investigate the signaling pathway that is involved in its actions in MC3T3-E1 cells. Methods: Cell proliferation, differentiation and mineralization were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, ALP test and Alizarin Red S staining, respectively. The concentration of sweroside in intracellular and extracellular fluids was determined by ultra-performance liquid chromatography coupled to triple quadrupole xevo-mass spectrometry (UPLC/TQ-XS-MS). Proteins associated with the osteoblastic signaling pathway were analysed by western blot and immunofluorescence methods. Results: Sweroside did not obviously affect the proliferation but significantly promoted the ALP activity and mineralization of MC3T3-E1 cells. The maximal absorption amount 0.465 ng/ml (1.3 x 10(-9) M) of sweroside was extremely lower than the tested concentration of 358.340 ng/ml (10(-6) M), indicating an extremely low absorption rate by MC3T3-E1 cells. Moreover, the ALP activity, the protein expression of ER-alpha and G protein-coupled receptor 30 (GPR30) induced by sweroside were markedly blocked by both the ER antagonist ICI 182780 and the GPR30 antagonist G15. In addition, sweroside also activated the phosphorylation of p38 kinase (p-p38), while the phosphorylation effects together with ALP and mineralization activities were completely blocked by a p38 antagonist, SB203580. Additionally, the phosphorylation of p38 induced by sweroside were markedly blocked by both the ER antagonist ICI 182780 and the GPR30 antagonist G15. Conclusions: The present study indicated that sweroside, as a potential agent in treatment of osteoporosis, might exert beneficial effects on MC3T3-E1 cells by interaction with the membrane estrogen receptor-alpha and GPR30 that then activates the p38 signaling pathway. This is the first study to report the specific mechanism of the effects of sweroside on osteoblastic differentiation and mineralization of MC3T3-E1 cells.
资助项目	State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica[SIMM1803KF-17] ; Shenzhen Basic Research Program[JCYJ20170818104152702] ; National Natural Science Foundation of China[81903616] ; Project of the Science Foundation for Distinguished Young Scholars of Guangdong Province[2014A030306043]
WOS关键词	ACTIVATED-PROTEIN-KINASE ; TRABECULAR BONE-FORMATION ; MAP KINASE ; ALKALINE-PHOSPHATASE ; ANDROGEN RECEPTOR ; UP-REGULATION ; PROLIFERATION ; EXPRESSION ; APOPTOSIS ; BETA
WOS研究方向	Plant Sciences ; Pharmacology & Pharmacy ; Integrative & Complementary Medicine
语种	英语
出版者	ELSEVIER GMBH
WOS记录号	WOS:000525848500006
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/281030]
专题	中国科学院上海药物研究所
通讯作者	Dai, Yi; Xiao, Hui-Hui
作者单位	1.Jinan Univ, Coll Pharm, Guangzhou 510632, Peoples R China 2.Jinan Univ, Int Cooperat Lab Tradit Chinese Med Modernizat &, Chinese Minist Educ, Guangzhou 510632, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China 4.Hong Kong Polytech Univ, State Key Lab Chinese Med & Mol Pharmacol Incubat, Shenzhen Res Inst, Shenzhen 518057, Peoples R China
推荐引用方式 GB/T 7714	Wu, Qing-Chang,Tang, Xi-Yang,Dai, Zi-Qin,et al. Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-alpha and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells[J]. PHYTOMEDICINE,2020,68:12.
APA	Wu, Qing-Chang,Tang, Xi-Yang,Dai, Zi-Qin,Dai, Yi,Xiao, Hui-Hui,&Yao, Xin-Sheng.(2020).Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-alpha and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells.PHYTOMEDICINE,68,12.
MLA	Wu, Qing-Chang,et al."Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-alpha and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells".PHYTOMEDICINE 68(2020):12.