| 谢婷婷1 杨乃龙1△ 徐丽丽1 王淑芳.尿酸影响人骨髓间充质干细胞成骨分化过程中BMP-2 表达[J].,2015,15(12):2251-2256 |
| 尿酸影响人骨髓间充质干细胞成骨分化过程中BMP-2 表达 |
| Effect of UA on the Expression of BMP-2 during the OsteogenicDifferentiation of Human Bone Marrow Mesenchymal StemCells |
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| DOI： |
| 中文关键词: 尿酸 人骨髓间充质干细胞 成骨分化 骨形态形成蛋白-2 |
| 英文关键词: UA Bone marrow mesenchymal stemcells, human Osteoblastic differentiation Bone morphogenetic protein-2 |
| 基金项目:国家自然科学基金项目（30871192）；青岛市科技基金项目(08-2-1-5-nsh-6) |
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| 中文摘要: |
| 目的：探讨在人骨髓间充质干细胞（hBMSCs）成骨分化过程中，不同浓度尿酸（UA）对骨形态形成蛋白-2（BMP-2）表达的影
响。方法：以全骨髓贴壁培养法分离hBMSCs，将生长状态良好的第3 代hBMSCs 分为5 组，分别为空白对照组（加入完全培养
基）和成骨诱导组（加入成骨诱导液及含0 mmol/L、0.2 mmol/L、0.4 mmol/L、0.8 mmol/L 尿酸的完全培养基）。连续干预诱导14d
后，用倒置显微镜观察细胞形态的变化，通过观察茜素红染色情况及检测碱性磷酸酶(ALP)活性进行成骨情况的检测。RT-PCR技
术检测各组细胞BMP-2 mRNA的表达情况。结果：第3 代hBMSCs大多为形态单一的长梭形，呈旋涡状生长；干预诱导后的细胞
逐渐变成不规则的立方形，局部形成团块状结节, 以含尿酸浓度为0.8 mmol/L的成骨诱导培养基最为显著。连续干预14d 后，空
白对照组茜素红染色为阴性，而各成骨诱导组细胞茜素红染色结果为阳性，提示干预诱导后的细胞为成骨细胞。碱性磷酸酶活性
随尿酸浓度的增加和干预时间的延长而增强（P<0.05）。RT-PCR 检测结果显示，空白对照组无BMP-2 mRNA的表达。成骨诱导组
随培养基中尿酸浓度的增加，BMP-2 mRNA表达逐渐增强，呈浓度依赖性（P < 0.05）。结论：尿酸上调hBMSCs 向成骨细胞分化过
程中BMP-2 mRNA的表达。 |
| 英文摘要: |
| Objective:To observe effect of uric acid (UA) on expression of bone morphogenetic protein-2 (BMP-2) in
differentiation process from human bone marrow mesenchymal stem cells (hBMSCs) to osteoblasts in vitro.Methods:hBMSCs were
isolated and cultured using the whole bone marrow adherence method. Passage 3 hBMSCs were divided into 5 groups: blank control
group (adding complete medium) and osteogenic induction group (adding osteoblast inducing media and 0 mmol/L, 0.2 mmol/L, 0.4
mmol/L, 0.8 mmol/L UA in complete medium). After 14 days of induction, cell morphology was observed under an inverted microscope.
Osteogenic ability was identified by alkaline phosphatase (ALP) activity assay and alizarin red stain. BMP-2 mRNA expression was
detected by reverse transcription PCR (RT-PCR).Results:Passage 3 cells were single long-spindle in shape and gradually formed a
whirlpool-shaped arrangement. After induction, the majority cells were from long-spindle to irregular cube in shape, and gradually
became paving stone or formed nodules. Among all the groups, the cells in 0.8 mmol/L UA induction medium formed the most nodules.
After 14 days of induction, calcium nodules in the blank control group were negtive, however, calcium nodules were dyed orange in
osteogenic induction group. The result showed hBMSCs were induced into osteoblasts successfully. ALP activity enhanced gradually
with the increase of intervention time and UA concentration (P<0.05). RT-PCR results showed that BMP-2 mRNA was hardly detected
in control group. In induced group, the mRNA expression of BMP-2 was evaluated with the increase of UA concentrations (P<0.05).Conclusion:UA can upregulate expression of BMP-2 mRNA during osteogenic differentiation of hBMSCs. |
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