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

Effect of polygonatum polysaccharides on the expression of ALP and BGP in the differentiation of the osteoblast by mouse bone marrow derived mesenchymal stem cells

目的探讨黄精多糖(PSP)对小鼠骨髓间充质干细胞向成骨细胞分化作用中碱性磷酸酶(alkaline phosphatase,ALP)和骨钙素(bone gla protein,BGP)表达的影响。方法取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7组用于实验。诱导组细胞采用体积分数为0.1双抗、含不同浓度PSP的血清+IMDM培养,非诱导组细胞采用体积分数为0.1双抗血清+IMDM培养。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。在培养第3,5,7,9,11,13天用四唑盐法(MTT)检测细胞的生长情况并绘制细胞生长曲线。在培养第7天、第14天采用ELISΑ酶联免疫吸附法分别测量细胞碱性磷酸酶(ALP)和骨钙素(BGP)的表达量。结果①小鼠骨髓间充质干细胞经PSP诱导后呈三角形、多角形、不规则状,可重叠生长。②各组细胞生长曲线总体趋势基本相同,在第7,9,11,13天,非诱导组细胞不再生长,而各PSP诱导组细胞均继续生长。③各不同浓度PSP诱导组均比非诱导组高表达ALP和BGP(P<0.01)。结论PSP可显著促小鼠骨髓间充质干细胞向成骨分化过程中ALP和BGP的表达,且随着浓度的增高该促进作用逐渐增强。

Objective To observe the effect of polygonatum polysaccharides （PSP） on the expression of alkaline phosphatase （ALP） and bone gla protein （BGP） in the differentiation of the osteoblast by mouse bone marrow derived mesenchymal stem cells （BMSCs）. Methods A BALB/C male 8-week mouse was selected. The marrow from the femur and the tibia was collected through sterile operation. The 3rd passage cells were collected and divided into 7 groups for the experiment. The cells in induced groups were cultured with 10% penicillin and streptomycin serum, additioned with different concentrations of PSP ＋ IMDM, while cells in non-induced group were cultured with 10% penicillin and streptomycin serum ＋ IMDM. The proliferation and morphologic changes of these cells were observed using inverted microscope everyday. At the 3rd, 5th, 7th, 9th, llth, and the 13th day of culturing, the proliferation and growth of cells were determined using MTT test. The expression of ALP and BGP was detected using ELISA kit at the 7th day and the 14th day. Results After the induction with PSP, BMSCs presented as triangle, astero-form, polygon, or irregular shape. Clumps and multilayer of cells became evident. The growth curves of cells in each group were roughly the same. At the 7th, 9th, 11 th, and the 13th day, ceils in non-induced group stopped growing, while the cells in induced groups kept growing. The expression of ALP and BGP in induced groups with different concentrations of PSP was higher than that in the non-induced group. Conclusion PSP can enhance the expression of ALP and BGP during the osteogenic differentitation of BMSCs, especially in a high concentration of PSP.