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兵豆氨酸促进人骨髓间充质干细胞的成骨分化和增殖。

Cajanine promotes osteogenic differentiation and proliferation of human bone marrow mesenchymal stem cells.

机构信息

Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine, China.

Cancer Center, 1st Hospital of Jilin University, Changchun, China.

出版信息

Adv Clin Exp Med. 2019 Jan;28(1):45-50. doi: 10.17219/acem/76638.

Abstract

BACKGROUND

Seed cells - mesenchymal stem cells (MSCs) - appear to be an attractive tool in the context of tissue engineering. Bone marrow represents the main source of MSCs for both experimental and clinical studies. However, the number limitation of bone marrow MSCs (BMSCs) and decreased function caused by proliferation make the search for adequate alternative sources of these cells for autologous and allogenic transplant necessary.

OBJECTIVES

This study was aimed to investigate the roles of cajanine isolated from the extracts of Cajanus cajan L. Millsp. in the proliferation and differentiation of BMSCs, and to discover the mechanism of proliferation of BMSCs promoted by cajanine.

MATERIAL AND METHODS

Bone marrow mesenchymal stem cells were cultured in high-glucose Dulbecco's Modified Eagle's Medium (DMEM) and osteogenic differentiation was induced by adding dexamethasone, ascorbic acid and β-glycerophosphate supplements. Bone marrow MSCs were cultured in medium without cajanine or supplemented with cajanine. The information about the proliferation and osteogenic differentiation of BMSCs was collated. The osteogenic differentiation potential of BMSCs was also assessed at the 3rd passage by Von Kossa staining. To observe cell signal transduction changes of BMSCs after culturing them with cajanine for 24 h, the western blot analysis was performed to detect phosphorylated cell cycle proteins and activated cyclins.

RESULTS

After osteogenic induction, the differentiation of BMSCs was accelerated by cajanine treatment. Osteogenesis markers were upregulated by cajanine treatment at both protein and mRNA levels. Cajanine obviously promoted the proliferation of BMSCs. After BMSCs were cultured with cajanine for 24 h, the cell cycle regulator proteins were phosphorylated or upregulated.

CONCLUSIONS

Cajanine can promote the expansion efficiency of BMSCs, at the same time keeping their multi-differentiation potential. Cajanine can activate the cell cycle signal transduction pathway, thus inducing cells to enter the G1/S phase and accelerating cells entering the G2/M phase. This study can contribute to the development of cajanine-based drugs in tissue engineering.

摘要

背景

种子细胞——间充质干细胞(MSCs)——在组织工程学中似乎是一种很有吸引力的工具。骨髓是实验和临床研究中 MSCs 的主要来源。然而,骨髓间充质干细胞(BMSCs)数量有限,增殖导致功能下降,这使得有必要寻找足够的替代来源,用于自体和同种异体移植。

目的

本研究旨在探讨从 Cajanus cajan L. Millsp. 提取物中分离得到的鹰爪豆碱对 BMSCs 增殖和分化的作用,并揭示鹰爪豆碱促进 BMSCs 增殖的机制。

材料和方法

在高糖 DMEM 中培养骨髓间充质干细胞,并通过添加地塞米松、抗坏血酸和β-甘油磷酸盐补充剂诱导成骨分化。骨髓间充质干细胞在无鹰爪豆碱或添加鹰爪豆碱的培养基中培养。收集关于 BMSCs 增殖和成骨分化的信息。在第 3 代时,通过 Von Kossa 染色评估 BMSCs 的成骨分化潜能。为了观察 BMSCs 培养 24 小时后细胞信号转导的变化,通过 Western blot 分析检测磷酸化细胞周期蛋白和激活的细胞周期蛋白。

结果

成骨诱导后,鹰爪豆碱处理加速了 BMSCs 的分化。鹰爪豆碱处理上调了成骨标志物的蛋白和 mRNA 水平。鹰爪豆碱明显促进了 BMSCs 的增殖。BMSCs 培养 24 小时后,细胞周期调控蛋白被磷酸化或上调。

结论

鹰爪豆碱能提高 BMSCs 的扩增效率,同时保持其多向分化潜能。鹰爪豆碱能激活细胞周期信号转导通路,从而诱导细胞进入 G1/S 期,并加速细胞进入 G2/M 期。本研究可为基于鹰爪豆碱的组织工程药物的开发提供参考。

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