银杏叶提取物对大鼠骨髓间充质干细胞成脂分化能力的影响

doi:10.13481/j.1671-587x.20150227

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (02): 343-346.doi: 10.13481/j.1671-587x.20150227

银杏叶提取物对大鼠骨髓间充质干细胞成脂分化能力的影响

谷旭¹, 方鸿满², 时舒曼¹, 张甲第¹, 吴哲¹

1. 吉林大学口腔医院口腔修复科, 吉林长春 130021;
2. 武警吉林省总队医院口腔科, 吉林长春 130052

收稿日期:2014-11-30 出版日期:2015-03-28 发布日期:2015-04-04
通讯作者: 吴哲, 教授, 硕士研究生导师(Tel:0431-88796018, E-mail:lcwztt@sina.com.cn) E-mail:lcwztt@sina.com.cn
作者简介:谷旭(1983-), 男, 吉林省四平市人, 主治医师, 在读医学硕士, 主要从事口腔修复和组织工程学方面的研究。
基金资助:
吉林省科技厅自然科学基金资助课题(201215052);吉林省科技厅科技发展计划项目资助课题(20140520051JH);吉林省发改委科研项目资助课题(2013C023-5)

Effect of Ginkgo biloba extract on adipogenic differentiation ability of bone marrow mesenchymal stem cells in rats

GU Xu¹, FANG Hongman², SHI Shuman¹, ZHANG Jiadi¹, WU Zhe¹

1. Department of Prosthodontics, Stomatology Hospital, Jilin University, Changchun 130021, China;
2. Department of Stomatology, Armed Police Military Hospital, Jilin Province, Changchun 130052, China

Received:2014-11-30 Online:2015-03-28 Published:2015-04-04

摘要/Abstract

摘要：

目的:探讨银杏叶提取物(GBE)对大鼠的骨髓间充质干细胞(BMMSCs)成脂分化能力的影响,阐明GBE在细胞水平上对骨质疏松治疗及预防的意义。方法:将50、100、150、200和400 mg·L^-1GBE与成脂诱导液加入到已纯化的第3代BMMSCs中(50、100、150、200和400 mg·L^-1GBE组),不加药的BMMSCs为对照组,共培养7 d,观察BMMSCs的形态学和表面抗原CD44、CD105和CD34的表达;采用油红O染色观察BMMSCs中脂滴形成情况,并对BMMSCs的成脂能力进行定量分析;采用RT-PCR法检测BMMSCs中脂肪组织脂肪酸结合蛋白(AP2)和过氧化物酶体增殖物激活受体(PPAR)的亚型(PPAR-γ)mRNA的表达水平。结果:形态学检测,体外培养的原代BMMSCs 3 d后可见细胞呈梭形,7 d后细胞呈漩涡状生长,同向排列;表面抗原标记物染色,CD44和CD105呈阳性表达,而CD34呈阴性表达,证明其为BMMSCs;油红O染色,约7 d后显微镜下见大量脂滴形成,并随着GBE浓度的增大脂滴数量减少;与对照组比较,其他各组BMMSCs的A₅₁₀值降低(P<0.05);RT-PCR检测,与对照组比较,其他各组细胞中AP2和PPAR-γ mRNA表达水平降低(P<0.05)。结论:GBE在体外可抑制BMMSCs的成脂分化能力,且抑制能力随着GBE浓度的增高而逐渐增强。

关键词: 银杏叶提取物, 骨髓间充质干细胞, 成脂分化, 骨质疏松

Abstract:

Objective To study the effect of Ginkgo biloba extract (GBE) on the adipogenic differentiation ability of the bone marrow mesenchymal stem cells(BMMSCs),and to clarify its significance in treatment and prevention of osteoporosis at the cell level.Methods The third generation of BMMSCs were cultured with adipogenic induction solution(control group),and 50,100,150,200, and 400 mg·L^-1 GBE(50,100,150,200, and 400 mg·L^-1 GBE groups) for 7 d.The morphology of BMMSCs and the expressions of surface antisgens CD44,CD105, and CD34 were observed;the formation of lipid droplet in the BMMSCs was observed by Oil Red O staining;the quantitative analysis of the adipogenic ability of the BMMSCs was performed;RT-PCR method was used to test the expression levels of fatty acid-binding protein(AP2) and peroxisome proliferator activated receptor-γ(PPAR-γ) mRNA.Results The morphological detection results showed that the BMMSCs were spindle shaped after cultured in vitro for 3 d,and grew in whorls and arranged in the same direction after cultured for 7 d.The surface antigens CD44 and CD105 showed positive expression,and CD34 showed negative expression,which proved the BMMSCs were extracted correctly;the Oil red O staining results showed a large of lipid droplets which was seen under microscope,and the number of lipid droplets was reduced with the increasing of the concentrations of GBE; compared with control group,the A₅₁₀ values of the BMMSCs in other groups were decreased(P<0.05).The RT-PCR results showed the expression levels of AP2 and PPAR-γ mRNA were decreasd in other groups compared with control group (P<0.05).Conclusion GBE can inhibit the adipogenic differentiation ability of BMMSCs in vitro,and the inhibitory ability is enhanced with the increasing of GBE concentration.

Key words: Ginkgo biloba extract, bone marrow mesenchymal stem cells, adipogenic differentiation, osteoporpsis

中图分类号:

R914

谷旭, 方鸿满, 时舒曼, 张甲第, 吴哲. 银杏叶提取物对大鼠骨髓间充质干细胞成脂分化能力的影响[J]. 吉林大学学报(医学版), 2015, 41(02): 343-346.

GU Xu, FANG Hongman, SHI Shuman, ZHANG Jiadi, WU Zhe. Effect of Ginkgo biloba extract on adipogenic differentiation ability of bone marrow mesenchymal stem cells in rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 343-346.

参考文献

[1] Lane JM,Russell L,Khan SN. Osteoporosis[J]. Clin Orthop Relat Res,2000(372):139-150.

[2] Moerman EJ,Teng K,Lipschitz DA.Aging activates adipogenic and suppresses osteogenic programs in mesenchymal marrow stroma/stem cells:the role of PPAR-gamma2 transcription factor and TGF-beta/BMP signaling pathways[J].Aging Cell,2004,3(6):379-389.

[3] Oh SM,Chung KH.Antiestrogenic activities of Ginkgo biloba extracts[J].J Steroid Biochem Mol Biol, 2006,100(4/5):167-176.

[4] Rodríguez JP,Astudillo P,Ríos S.Adipogenesis and osteoporosis[J].Rev Méd Chile,2009,137(6):827-836.

[5] Oh SM,Kim HR,Chung KH.Effects of ginkgo biloba on in vitro osteoblast cells and ovariectomized rat osteoclast cells[J].Arch Pharm Res,2008,31(2):216-224.

[6] Trivedi R,Kumar A,Gupta V.Effects of Egb 761 on bone mineral density,bone microstructure,and osteoblast function:Possible roles of quercetin and kaempferol[J].Mol Cell Endocrinol,2009,302(1):86-91.

[7] Park SH,Sim WY,Park SW,et al.An electromagnetic compressive force by cell exciter stimulates chondrogenic differentiation of bone marrow-derived mesenchymal stem cells[J]. Tissue Eng,2006,12(11):3107-3117.

[8] de Mattos Carvalho A,Alves AL,Golim MA,et al.Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue[J]. Vet Immunol Immunopathol,2009,132(2-4):303-306.

[9] Kode JA,Mukherjee S,Joglekar MV,et al. Mesenchymal stem cells:immunobiology and role in immunomodulation and tissue regeneration[J]. Cytotherapy,2009,11(4):377-391.

[10] Lei Z,Yongda L,Jun M,et al.Culture and neural differentiation of rat bone marrow mesenchymal stem cells in vitro[J]. Cell Biol Int,2007,31(9):916-923.

[11] Beyer Nardi N,da Silva Meirelles L.Mesenchymal stem cells:isolation,in vitro expansion and characterization[J]. Handb Exp Pharmacol,2006(174):249-282.

[12] Morikawa S,Mabuchi Y,Kubota Y,et al.Prospective identification,isolation,and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow[J]. J Exp Med,2009,206(11):2483-2496.

[13] Agorogiannis GI,Alexaki VI,Castana O,et al.Topical application of autologous adipose-derived mesenchymal stem cells (MSCs) for persistent sterile corneal epithelial defect[J].Graefes Arch Clin Exp Ophthalmol,2012,250(3):455-457.

[14] Arufe MC,De la Fuente A,Fuentes I,et al.Chondrogenic potential of subpopulations of cells expressing mesenchymal stem cell markers dericed from human synovial membrances[J]. J Cell Biochem,2010,111(4):834-845.

[15] Heidari B,Shirazi A,Akhondi MM,et al. Comparison of proliferative and multilineage differentiation potential of sheep mesenchymal stem cells derived from bone marrow,liver,and adipose tissue[J].Avicenna J Med Biotechnol,2013,5(2):104-117.

[16] James AW. Review of signaling pathways governing MSC osteogenic and adipogenic differentiation[J].Scientifica (Cairo),2013,2013:684736.doi:10.1155/2013/684736.

[17] Gao B,Huang Q,Lin YS,et al.Dose-dependent effect of estrogen suppresses the osteo-adipogenic transdifferentiation of osteoblasts via canonical Wnt signaling pathway[J].PLoS One,2014,9(6):e99137.

[18] 潘孝聪,张兴彩.银杏叶提取物治疗肺间质纤维化[J].吉林中医药,2013,33(11):1103-1105.

[19] 王新月,张倩,张永和,等.中药有效成分抗心肌缺血作用概述[J].吉林中医药,2013,33(6):640-642.

银杏叶提取物对大鼠骨髓间充质干细胞成脂分化能力的影响

Effect of Ginkgo biloba extract on adipogenic differentiation ability of bone marrow mesenchymal stem cells in rats

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