Polygonatum sibiricum polysaccharide promotes osteogenesis by signaling pathway activation after LRP5 silencing

doi:10.3969/j.issn.2095-4344.2017.04.001

Abstract

Abstract:

Abstract
BACKGROUND: Our previous studies have found that polygonatum sibiricum polysaccharide (PSP) promotes osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by Wnt/β-catenin signaling pathway, but the molecular mechanism is unclear.
OBJECTIVE: To investigate the effect of PSP promoting the osteogenic differentiation via Wnt signaling pathways in BMSCs after LRP5 silencing.
METHODS: LRP5 interference vectors were constructed and then transfected into C57BL/6 mouse BMSCs cultured in vitro. The transfection efficiency of cells was calculated under fluorescence inverted microscope and the expression of LRP5 protein was detected by western blot assay. The osteogenic potential of BMSCs after LRP5-siRNA transfection was analyzed by alkaline phosphatase staining, alizarin red staining and western blot assay. Effect of PSP on the osteogenic differentiation of LIRP5-silenced mouse BMSCs was detected by real-time PCR and dual luciferase assay.
RESULTS AND CONCLUSION: Compared with the control group, the mineralization ability, the mRNA expressions of Runx2 and Osterix, and the protein expression of LRP5 were significantly decreased in the LRP5-siRNA group (P < 0.05). PSP could promote LRP5-siRNA transfected mouse BMSCs differentiating into osteoblasts and significantly upregulated the expressions of β-catenin and Osterixin, and also induced the high expression of luciferase reporter gene (TOPFlash) containing wild type TCF binding sites (P < 0.05). To conclude, LRP5 plays an important role in the process of mouse BMSCs differentiating into osteoblasts. PSP can promote the osteogenic differentiation of mouse BMSCs by activating the Wnt/β-catenin signaling pathway independent on LRP5.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Polygonatum, Membrane Proteins, Cell Differentiation, Osteoporosis, Tissue Engineering

CLC Number:

R318

Peng Xiao-ming1, Zong Shao-hui2, Zeng Gao-feng3, Nong Meng-ni3, Du Li1, Li Ke-ke1, He Ji-chen1, Shi Xiong-zhi1, Wu Yun-le1. Polygonatum sibiricum polysaccharide promotes osteogenesis by signaling pathway activation after LRP5 silencing[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(4): 493-498.

Figures/Tables 3

References

[1] Neer RM, Arnaud CD, Zanchetta JR, et al. Effect of parathyroid hormone (1-34) on fractures and bone mineral density in postmenopausal women with osteoporosis.N Engl J Med. 2001;344(19):1434-1441.

[2] Tanaka Y, Nakayamada S, Okada Y.Osteoblasts and osteoclasts in bone remodeling and inflammation.Curr Drug Targets Inflamm Allergy.2005;4(3):325-328.

[3] Black DM, Delmas PD, Eastell R, et al. Once-yearly zoledronic acid for treatmentof postmenopausal osteoporosis.N Engl J Med. 2007;356(18):1809-1822.

[4] Bock O, Boerst H, Thomasius F E, et al. Common musculoskeletal adverse effects of oral treatment with once weekly alendronate and risedronate in patients with osteoporosis and ways for their prevention.J Musculoskelet Neuronal Interact.2007;7(2):144-148.

[5] Mchorney CA, Schousboe JT, Cline RR, et al.The impact of osteoporosis medication beliefs and side-effect experiences on non-adherence to oral bisphosphonates.Current Medical Research & Opinion.2007;23(12):3137-3152.

[6] Inderjeeth CA, Lee S, Glendenning P. Efficacy, side-effects and route of administration are more important than frequency of dosing of anti-osteoporosis treatments in determining patient preference and compliance: A critical review of published articles from 1970-2009. Bone. 2010;47(Suppl 1):S206.

[7] 雷震,杨光义,叶方,等.黄精多糖药理作用及临床应用研究概述[J].中国药师,2012,(01):114-116.

[8] 何才通,李文.黄精多糖药理功效研究进展[J].新中医,2014(03): 196-199.

[9] 胡敏,王琴,周晓东,等.黄精药理作用研究进展及其临床应用[J].广东药学, 2005,(05):68-71.

[10] 曾高峰,宗少晖,邹斌，等.黄精多糖对小鼠骨髓间充质干细胞向成骨细胞分化中ALP和BGP表达的影响[J].中国骨质疏松杂志, 2014,(7):779-783.

[11] 农梦妮,曾高峰,宗少晖,等.黄精多糖调控骨髓间充质干细胞向成骨细胞分化[J].中国组织工程研究,2016,20(15):2133-2139.

[12] Hadjidakis DJ, Androulakis II. Bone Remodeling.Ann N Y Acad Sci. 2006;1092:385-96.

[13] Ferron M,Wei J,Yoshizawa T,et al.Insulin Signaling in Osteoblasts Integrates Bone Remodeling and Energy Metabolism. Cell.2010; 142(2):296-308.

[14] Pluskiewicz W, Adamczyk PE, Leszczynski P, et al. Ten-year probability of osteoporotic fracture in 2012 Polish women assessed by FRAX and nomogram by Nguyen et al.- Conformity between methods and their clinical utility. Bone. 2010;46(46):1661-1667.

[15] Bliuc D,Nguyen ND,Milch VE, et al. Mortality risk associated with low-trauma osteoporotic fracture and subsequent fracture in men and women.JAMA.2009;301(5):513-521.

[16] Lippuner K, Johansson H, Kanis J A, et al. Remaining lifetime and absolute 10-year probabilities of osteoporotic fracture in Swiss men and women. Osteoporos Int. 2009;20(7): 1131-1140.

[17] Khang D, Choi J, Im YM, et al. Role of subnano-, nano- and submicron-surface features on osteoblast differentiation of bone marrow mesenchymal stem cells. Biomaterials.2012; 33(26):5997-6007.

[18] Kemp KC, Hows J, Donaldso DC. Bone Marrow-Derived Mesenchymal Stem Cells. Leuk Lymphoma. 2005;46(11): 1531-1544.

[19] Fata JE, Kong YY, Li J, et al.The osteoclast differentiation factor osteoprotegerin-ligand is essential for mammary gland development.Cell. 2000;103(1): 41-50.

[20] Ponte AL, Marais E, Gallay N, et al. The In Vitro Migration Capacity of Human Bone Marrow Mesenchymal Stem Cells: Comparison of Chemokine and Growth Factor Chemotactic Activities.Stem Cells.2007;25(7):1737-1745.

[21] Zhou S, Greenberger J, Epperly M, et al. Age-related intrinsic changes in human bone-marrow-derived mesenchymal stem cells and their differentiation to osteoblasts.Aging Cell.2008；7(3):335-343.

[22] Darnay BG, Ni J, Moore PA, et al. Activation of NF-kappaB by RANK requires tumor necrosis factor receptor-associated factor (TRAF) 6 and NF-kappaB-inducing kinase.J Biol Chem. 1999;274(12):7724-7731. .

[23] Jarrett H,Soles E,Sirna S,et al. Risk Factor Predominance in Atherosclerotic Cardiovascular Disease Differs by Race: An Observational Study. J Clin Lipidology.2015; 9(3):423.

[24] 曾高峰,宗少晖,邹斌,等.黄精多糖对小鼠骨髓间充质干细胞向成骨细胞分化中PINP和BMP-2表达的影响[J].天然产物研究与开发,2014,(08): 1188-1192.

[25] Natsag J,Kendall MA,Sellmeyer DE,et al. Vitamin D, osteoprotegerin/receptor activator ofnuclear factor-kappaBligand (OPG/RANKL) and inflammation with alendronate treatment in HIV-infected patients with reduced bone mineral density.HIV Med. 2016;17(3):196-205.