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| 贝尼地平对小鼠骨髓间充质干细胞成骨分化体外研究 |
| Effect of benidipine on osteogenic differentiation of bone marrow stromal cells in vitro |
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| DOI：10.3969/j.issn.1006.7108.2016.04.008 |
| 中文关键词: 贝尼地平 成骨；骨髓间充质干细胞 抗骨质疏松 钙通道阻滞剂 |
| 英文关键词:Benidipine Osteogenesis BMSCs Anti-osteoporosis Calcium Channel Blocker |
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| 中文摘要: |
| 目的 在体外环境下探讨贝尼地平对小鼠骨髓间充质干细胞向成骨细胞分化的影响。方法 C57/BL6小鼠骨髄间充质干细胞（bone marrow stromal cells，BMSCs)在成骨诱导培养基条件下经过1μmol/L、10μmol/L和100μmol/L浓度的贝尼地平（BD)处理，1天和14天后采用CCK8法检测细胞增殖情况，以及14天后采用碱性磷酸酶（ALP)染色检测ALP表达量，蛋白免疫印迹（Western Bloting)法检测成骨特征蛋白骨钙素（OCN)和核心结合蛋白因子2( Runx2)以及WNT/β-catenin信号通路关键蛋白β-链蛋白（β-catenin)和低密度脂蛋白相关蛋白5(LRP5)蛋白的表达。结果 在本次研究中，贝尼地平浓度在1 ~ 100 μmol/L对细胞没有毒性作用（P>0.05);所有浓度的贝尼地平处理组（1?100μmol /L)中ALP表达量均增加（P <0. 05), 并出现浓度依赖性，随着贝尼地平浓度的增加促进BMSCs表达ALP能力相应增强（P<0.05);贝尼地平促进所有处理组（1 ~ 100 μmol/L)中Runx2的表达（P<0.05),并出现浓度依赖性，在贝尼地平浓度为100 μmol/L时促进作用最强（P< 0. 05);OCN的表达随着贝尼地平浓度的增加而相应增加（P<0, 05),在贝尼地平浓度为100 μmol/L时达到最大（P<0.05);贝尼地平促进β-catenin表达（P<0.05),出现随贝尼地平浓度增加而表达增强，促进作用在贝尼地平浓度为100 μmol/L时达到最大 (P<0.05);而LRP5的表达在贝尼地平的作用下得到加强（P<0. 05)并出现浓度依赖性，在贝尼地平浓度为100 μmol/L时达到最大（P <0.05)。结论 贝尼地平在体外能通过上调WNT/β-catenin信号通路活性促进骨髓间充质干细胞向成骨细胞分化,这对髙血压合并骨质疏松患者而言是一个良好的治疗药物。 |
| 英文摘要: |
| Objective To investigate the effect of benidipine of osteoblast differentiation by mouse bone marrow stromal cells in vitro. Methods Under osteogenic conditions, bone marrow stromal cells (BMSCs) from C57/BL6 mice were cultured for 2 weeks. The cultures were additioned with 0, 1, 10, 100 μmol/L of benidipine. Cell proliferation was examined with CCK8 assays on 24 hours and 2 weeks. Protein expressions of alkaline phosphatase (ALP) , osteocalcin (OCN), runt-related transcription factor 2 (Runx2), β-catenin, and low-density lipoprotein receptor-related protein 5 (LRP5) were evaluated on 14 days. Results In our study, benidipine at concentrations of 1-100 μM did not significantly affect cell growth after treatment (P<0. 05). The expression of ALP increased in cultures with benidipine (1-100 μmol/L) in a dose dependent manner (P < 0. 05). Runx2 was also up- regulated with benidipine (1-100 μmoI/L) treatment ( P < 0. 05 ) in a dose dependent manner, and the peak of Runx2 expression was found at 100 μmol/L treatment (P < 0. 05). The expression of OCN, β-catenin, and LRP5 increased in cultures with benidipine ( 1-100 μmol/L) in a dose dependent manner (P<0. 05), and the peak of the expression was found at 100 μmol/L (P <0.05). Conclusion Benidipine promotes BMSCs differentiation into osteoblasts through up regulation of WNT/β-catenin signal pathway. Therefore, it might be a suitable candidate for the treatment of patients with osteoporosis and hypertension. |
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