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

天然产物研究与开发 ›› 2014, Vol. 26 ›› Issue (8): 1188-1192.

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

曾高峰1*，宗少晖2，邹斌3，李柯柯4

1 广西医科大学公共卫生学院；2 广西医科大学第一附属医院脊柱骨病外科；3 广西医科大学七年制临床医学2007级研究生；4 广西医科大学研究生学院，南宁 530021

出版日期:2014-08-30 发布日期:2014-11-06

The Expression of BMP-2 and PINP in the Osteoblast Differentiation of Mouse Bone Marrow Derived Mesenchymal Stem Cell Induced by Polygonatum Polysaccharide

ZENG Gao-feng1*, ZONG Shao-hui2, ZOU Bin3, LI Ke-ke4

1 College of Public Hygiene of Guangxi Medical University；2 Department of Osteopathia,the First Affiliated Hospital of Guangxi Medical University；3 The sevenyear combined Bachelor’s and Master’s Degree Program of clinical medicine of Guangxi Medical University; 4 Graduate School of Guangxi Medical University, Guangxi Nanning 530021 , P.R.China

Online:2014-08-30 Published:2014-11-06

摘要/Abstract

摘要： 探讨黄精多糖（PSP）对小鼠骨髓间充质干细胞向成骨细胞分化作用中I型前胶原N端前肽（Procollagen typeⅠN-terminal Propeptide, PINP）和骨形态发生蛋白2（Bone morphogenetic protein-2, BMP-2）表达的影响。取8周龄雄性BALB/C小鼠1只，无菌操作分离小鼠股骨和胫骨，注射器冲出骨髓制成细胞悬液，传代3次后分为6组用于实验。空白诱导组仅加入等量成骨诱导培养基；阳性对照诱导组加入等量成骨诱导培养基及雌二醇（10^-8 mol/L）；各PSP诱导组：加成骨诱导培养基和各自浓度的PSP（200、300、400、500 mg/mL）。每天在倒置显微镜下观察各组细胞的生长情况及形态变化。倒置相差显微镜镜下观察细胞形态变化。在培养第7 d、第14 d采用ELISA酶联免疫吸附法分别测量细胞PINP和BMP-2的表达量。实验结果显示小鼠骨髓间充质干细胞原代培养呈长梭形,有接触抑制现象,诱导后细胞呈三角形、多角形、不规则形状，细胞生长密集时可重叠生长。各不同浓度PSP诱导组均比非诱导组高表达PINP和BMP-2（P<0.01）。表明PSP呈剂量相关性促进体外培养的小鼠骨髓间充质干细胞成骨分化,高浓度PSP可显著促进骨髓间充质干细胞骨向分化过程中BMP-2和PINP的表达。

关键词: 黄精多糖, 骨髓间充质干细胞, 成骨细胞分化, Ⅰ型前胶原N端前肽, 骨形态发生蛋白2

Abstract: The objective of this study was to observe the effects of polygonatum polysaccharide on the osteogenic differentiation of murine bone marrow derived mesenchymal stem cells (BMSCs). One BALB/C male mouse aged 8 weeks was used for femoral and tibial marrow by sterile working.The third generation of the cells was divided in 6 groups for this experiment.The blank induced group was cultured in osteogenic medium (OM). The positive control induced group was cultured in OM with 10^-8 mol/L of 17β-E2.The PSP200-, PSP300-, PSP400- and PSP500-induced groups were cultured in OM with 200,300,400 and 500 mg/mL of PSP, respectively.Inverted microscope was used to observe the proliferation and morphologic change of these cells every day.The expression dose of PINP and BMP-2 were separately detected by ELISA kit at the 7th day and at the 14th day. After induced,BMSCs were proliferated effectively,presented triangle,asteroform,polygon or irregular shape.Clumps and multilayer of cells became evident.Compared to the non-induced group,the cells of PSP induced groups had a high expression dose of PINP and BMP-2 (P<0.01).This demonstrated that PSP can promote the proliferation of BMSCs and it can enhance the expression of PINP and BMP-2,especially in a high concentration of PSP.

Key words: Polygonatum polysaccharide, bone marrow derived mesenchymal stem cells, osteogenic differentiation, PINP, BMP-2

中图分类号:

R285.5

曾高峰1*，宗少晖2，邹斌3，李柯柯4. 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, 26(8): 1188-1192.

ZENG Gao-feng1*, ZONG Shao-hui2, ZOU Bin3, LI Ke-ke4. The Expression of BMP-2 and PINP in the Osteoblast Differentiation of Mouse Bone Marrow Derived Mesenchymal Stem Cell Induced by Polygonatum Polysaccharide[J]. NATURAL PRODUCT RESEARCH AND DEVELOPMENT, 2014, 26(8): 1188-1192.

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