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| 辛伐他汀通过p38MAPK信号通路诱导BMSCs成骨分化的研究 |
| Simvastatin induces osteogenic differentiation of BMSCs via p38MAPK pathway |
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| DOI：10.3969/j.issn.1006-7108.2016.02.001 |
| 中文关键词: 辛伐他汀 骨髓间充质干细胞 成骨分化 p38丝裂原活化激酶 |
| 英文关键词:Simvastatin Bone marrow mesenchymal stem cells Osteogenic differentiation p38 Mitogen-activated protein kinase |
| 基金项目:河北省自然科学基金（H2013209255）；河北省高等学校科学研究计划（QN20131007）；唐山市科科学技术研究与发展指令计划项目（13130281Z）；唐山市老年医学重点实验室资助项目（14140221B） |
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| 中文摘要: |
| 目的 观察辛伐他汀（SIM）体外单独给药对大鼠骨髓间充质干细胞（BMSCs）成骨分化的影响。方法 取第二代大鼠BMSCs随机分为4组，对照组（CM）：完全培养基培养；诱导组（OM）：成骨诱导培养基培养；辛伐他汀组（SIM）：含终浓度为10-7 mol/L的辛伐他汀的完全培养基培养；阻断剂组（SB+SIM）：先用p38MAPK通路阻断剂SB203580干预30min后，加入等浓度辛伐他汀。给药4h后，Western Blot检测p-p38MAPK、p38MAPK蛋白的表达。给药7d后，进行碱性磷酸酶（ALP）染色和比活性测定。给药7、14d后，采用Real-time PCR法检测ALP、Ι型胶原（COLΙ）、骨钙素（OCN）和Runt相关转录因子2（Runx2）mRNA的表达。结果 ①Western Blot检测：OM组和SIM组p-p38MAPK表达水平显著高于CM组和SB+SIM组（P＜0.05）。②ALP比活性测定：OM 组和SIM组显著高于CM组，与相比，SB+SIM组显著低于SIM组（P＜0.05）。③ALP染色：OM 组和SIM组细胞胞浆内出现棕色颗粒，而SB+SIM组和CM组未见明显阳性染色颗粒。④Real-time PCR检测：在第7d、14d，OM组和SIM组ALP、COLΙ 和Runx2 mRNA表达水平均显著高于CM组，SB+SIM组明显低于SIM组（P＜0.05）；第7d，OM组OCN mRNA表达水平明显高于CM组（P＜0.05）；在第14d，OM组和SIM组OCN mRNA表达水平高于CM组，SB+SIM组低于SIM组（P＜0.05）。结论 辛伐他汀可以在缺乏成骨诱导成分的环境下，通过活化p38MAPK信号诱导大鼠BMSCs向成骨细胞分化。 |
| 英文摘要: |
| Objective To observe the effect of simvastatin alone on osteogenic differentiation by rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods The second passage of rat BMSCs were randomly divided into 4 groups, CM group (complete medium in the culture), OM group (induction medium in the culture), SIM group (addition of 10-7 mol/L simvastatin in medium), and SB+SIM group (intervention with p38MAPK pathway blocker SB203580 for 30 min, and then addition of 10-7 mol/L simvastatin). Protein expressions of p-p38MAPK and p38MAPK were detected using Western blotting after 4h treatment. ALP staining and ALP activity were measured after 7d treatment. ALP, COLΙ, OCN, and Runx2 mRNA expressions were detected using real-time RT-PCR assay after 7d and 14d treatment. Results 1) Western blotting analysis showed that expression of p-p38MAPK level was significantly higher in OM and SIM groups than in CM and SB+SIM groups (P<0.05). 2) ALP activity was higher in OM group than in SIM group. It was lower in SB+SIM group than in SIM group (P<0.05). 3) ALP positive staining was shown in OM group and the SIM group, but not in SB+SIM group and CM group. 4) Real-time PCR analysis showed that on 7d and 14d, ALP, COLΙ and Runx2 mRNA expression levels in OM Group and SIM group were higher than those in CM group (P<0.05). They were significantly lower in SB+SIM group than in SIM group (P<0.05). OCN mRNA expression level was significantly higher in OM Group than in CM group on 7d (P<0.05). It was higher in OM group and SIM groups than in CM group on 14d (P<0.05). It was lower in SB+SIM group than in SIM group on 14d (P<0.05). Conclusion In the absence of osteogenic induction environment, simvastatin can induce differentiation of BMSCs into osteoblasts via p38MAPK pathway. |
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