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| GLP-1对GK糖尿病大鼠骨髓间充质干细胞骨向分化能力的影响 |
| Effect of GLP-1 on the osteogenic differentiation by bone mesenchymal stem cells in Goto-Kakizaki rats |
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| DOI：10.3969/j.issn.1006.7108.2015.03.010 |
| 中文关键词: 胰高血糖素样肽4 ( GLP-1) 骨向分化 信号通路 |
| 英文关键词:GLP-1 Osteogenic differentiation Signal pathway |
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| 中文摘要: |
| 目的 观察胰高血糖素样肽-l( Glucagon-like peptide-I，GLP-l)对Goto-Kakizaki鼠骨髓间充质干细胞（Bone marrow stromal cell，BMSCs)成骨分化能力的影响，并讨探其可能作用机制。方法 贴壁法取8周龄GK鼠BMSCs，逆转录PCR法验证BMSCs表达GLP-1受体，MTT法筛选GLP-1对GK鼠BMSCs的最佳刺激浓度，选取该浓度干预GK鼠BMSCs，观察其成骨分化相关指标：第7 d、14 d测定碱性磷酸酶活性，第14d应用实时定量PCR检测ALP、RUNX2、OCN、Smad1、β-catenin、OPG和 RANKL的表达。结果 1. GK鼠BMSCs中存在GLP-1受体;2. 20 nmol/L的GLP-1对细胞刺激作用最佳;3.成骨诱导后ALP 活性升高。成骨诱导7 d，诱导组和GLP-1干预组的ALP活性相对于对照组均明显升高(P <0. 05);成骨诱导14 d，诱导组和 GLP-1干预组的ALP活性进一步升高（与自身7d时相比，P <0.05)，且均高于对照组(P <0. 05); GLP-1干预组ALP活性在第 7d时稍高于诱导组(P >0. 05)，第14d时明显高于诱导组(P <0. 05) ；4.与正常成骨诱导组相比，GLP-1干预组ALP和RUNX 2表达量均明显增加(P <0.5)，OCN的表达量增加，但无统计学意义。GLP-1干预组表达Smad1减少（P > 0. 05)，β-catenin 明显增多（P <0.05)。GLP-1干预组表达 OPG增加（P >0.05)，RANKL 减少（P <0. 05)，OPG/RANKL 升高（P <0.05)。结论 1. GK鼠BMSCs上存在GLP-1受体;2. GLP-1可促进GK鼠BMSCs向成骨细胞分化，并调节Wnt通路部分基因的表达及OPG/RANK/RANKL轴的动态平衡。 |
| 英文摘要: |
| Objective To investigate the effect of glucagon-like peptide-L ( GLP-1) on the ability of osteoblast differentiation from bone marrow stromal cells (BMSCs)，and the possible mechanism. Methods BMSCs was extracted from 8-week-old GK rats. Real-time RT-PCR was used to evaluate the expression level of GLP-1 receptor. MTT assay was used to assess the best concentration of stimulation of GLP-1. GLP-1 at this concentration was used to induce cell differentiation from BMSCs to osteoblasts. ALP staining was performed on the 7th and 14th day of osteoblasts induction. Real-time RT-PCR was used to evaluate the expression levels of ALP，RUX2，OCN，Smadl，β-catenin，OPG，and RANKL on the 14th day. Results There were GLP- 1 receptors in BMSCs. The best concentration of cell stimulation with GLP-1 was 20 nmol/L. ALP activity increased after osteogenic induction. After 7d osteogenic induction，ALP activity in induction group and GLP-1 intervention group was significantly higher than that in the control group (P <0. 05). After 14d osteogenic induction，ALP activity in induction group and GLP-1 intervention group was even much higher than that in the control group and that on 7d (P <0. 05). The activity of ALP in GLP-1 group was slightly higher than that in induction group on the 7th day，but significantly higher on the 14th day (P <0. 05). The expression levels of ALP and RUNX2 increased markedly in GLP-1 group (P <0. 05). Although the expression level of OCN increased，there was no statistical significance. The expression level of Smad1 reduced in GLP-1 group (P <0.05). The expression level of β-catenin increased significantly (P <0.05). After the intervention of GLP-1，the expression level of OPG increased (P > 0.05)，the expression level of RANKL decreased (P < 0. 05)，and the expression level of OPG/RANKL increased (P < 0. 05). Conclusion There were GLP-1 receptors in BMSCs of GK rats. GLP-1 can promote the osteoblast differentiation from BMSCs and regulate the expression of some genes in Wnt signal pathway and influence the dynamic balance in OPG/RANK/RANKL axial. |
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