胰高血糖素样肽-1及其类似物EX-4促进大鼠骨髓间充质干细胞的增殖与成骨分化

中国临床药理学与治疗学 ›› 2015, Vol. 20 ›› Issue (2): 121-127.

• 基础研究 • 下一篇

胰高血糖素样肽-1及其类似物EX-4促进大鼠骨髓间充质干细胞的增殖与成骨分化

周诗萌¹, 王宁², 孟静茹², 贾敏², 胡静³, 马雪²

¹第四军医大学药学系学员队, 西安 710032,陕西;
²第四军医大学药学系药理学教研室,西安 710032, 陕西;
³第四军医大学基础部人体解剖与组织胚胎学教研室,西安 710032,陕西

收稿日期:2014-04-08 修回日期:2014-11-03 出版日期:2015-02-26 发布日期:2015-03-20
通讯作者: 马雪,女,博士,讲师,研究方向:代谢性疾病的防治。Tel: 029-84774555 E-mail: maxuel@fmmu.edu.cn
作者简介:周诗萌,女,本科,研究方向:代谢性疾病的防治。Tel: 029-84774555 E-mail: zsmapril_417@163.com
基金资助:
国家自然科学基金项目(81100626;81201515)

Glucagon-like peptide-1 and its analogue exendin-4 enhance proliferation and osteogenic differentiation of rat bone mesenchymal stem cells in vitro

ZHOU Shi-meng¹, WANG Ning², MENG Jing-ru², JIA Min², HU Jing³, MA Xue²

¹Department of Pharmacy;
²Department of Pharmacology;
³Department of Histology and Embryology, Fourth Military Medical University, Xi'an 710032, Shannxi, China

Received:2014-04-08 Revised:2014-11-03 Online:2015-02-26 Published:2015-03-20

摘要/Abstract

摘要： 目的: 观察胰高血糖素样肽-1(GLP-1)及其受体激动剂exendin-4(EX-4)对体外培养大鼠骨髓间充质干细胞(BMSCs)增殖与成骨分化的影响。方法: 取大鼠股骨和胫骨全骨髓,采用贴壁法分离BMSCs并传代至第三代,培养2 d后分成4组:BMSC组、诱导成骨组(osteoblast induced medium,OIM组)和诱导加药组,诱导加药组细分为GLP-1(10^-9~10^-7 mol/L)或EX-4(10^-9~10^-7 mol/L)的不同浓度组,MTT法检测细胞增殖能力;测定碱性磷酸酶(ALP)活性;通过ALP染色及茜素红染色观察药物诱导BMSCs向成骨细胞分化、矿化的效果;采用RT-PCR方法检测ALP和Runx2成骨相关基因的表达情况。结果: 与诱导对照组相比,GLP-1及EX-4各浓度处理组呈剂量依赖性显著促进细胞增殖(P<0.05);ALP染色与茜素红染色结果显示,对照组无显色或显色不明显,而GLP-1及EX-4组均有阳性显色。GLP-1及EX-4可以显著提高ALP活性,并提高Runx2、ALP的表达水平。结论: GLP-1及EX-4不仅能够直接促进BMSCs增殖,并促进其向成骨细胞分化,GLP-1及EX-4可能在骨重建中发挥重要的作用。

关键词: 胰高血糖素样肽-1, 骨髓间充质干细胞, 成骨性分化, 碱性磷酸酶

Abstract: AIM: To investigate the effects of GLP-1 and GLP-1R agonist exendin-4(EX-4) on the proliferation and differentiation of rat bone mesenchymal stem cells (BMSCs).METHODS: BMSCs were isolated from rats'tibia and femur by the method of density gradient centrifugation and attachment were passaged to the three generation. 4 groups were divided after 2 days cultivation, BMSC group, osteoblast induced medium(OIM),and drug induced medium. In the drug induced medium, different concentration of GLP-1 or EX-4 (10^-9-10^-7 mol/L) were added to the media of the BMSC for the observation of cell proliferation (OD₄₅₀),`differentiation (ALP staining) and mineralization (alizarin red staining for the observation of calcium nodule) of the BMSCs. RT-PCR was used to detect the expression of ALP, Runx2.RESULTS: Compared with the control group,GLP-1,EX-4 treatment groups promoted cells proliferation in a dose-dependent manner (P<0.05); the ALP and alizarin red staining suggested,GLP-1,EX-4 treatment groups were positive,the control group was weak stained or did not stain at all.ALP activity was improved, and the expressions of ALP and Runx2 were increased.CONCLUSION: The proliferation, differentiation and mineralization of the BMSCs are promoted directly by GLP-1 and EX-4, which may play an important role in bone reconstruction.

Key words: GLP-1, bone mesenchymal stem cells (BMSCs), osteogenesis, ALP

中图分类号:

R965.2

周诗萌, 王宁, 孟静茹, 贾敏, 胡静, 马雪. 胰高血糖素样肽-1及其类似物EX-4促进大鼠骨髓间充质干细胞的增殖与成骨分化[J]. 中国临床药理学与治疗学, 2015, 20(2): 121-127.

ZHOU Shi-meng, WANG Ning, MENG Jing-ru, JIA Min, HU Jing, MA Xue. Glucagon-like peptide-1 and its analogue exendin-4 enhance proliferation and osteogenic differentiation of rat bone mesenchymal stem cells in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(2): 121-127.

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胰高血糖素样肽-1及其类似物EX-4促进大鼠骨髓间充质干细胞的增殖与成骨分化

Glucagon-like peptide-1 and its analogue exendin-4 enhance proliferation and osteogenic differentiation of rat bone mesenchymal stem cells in vitro

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