文章摘要
张勇,关邯峰,方忠,等.氨来呫诺对大鼠骨髓间充质干细胞成骨分化的影响.骨科,2018,9(3): 229-234.
氨来呫诺对大鼠骨髓间充质干细胞成骨分化的影响
Effects of amlexanox on the osteogenic differentiation of rat bone marrow mesenchymal stem cells
投稿时间:2018-02-17  
DOI:10.3969/j.issn.1674-8573.2018.03.013
中文关键词: 氨来呫诺  骨髓间充质干细胞  成骨分化  骨质疏松
英文关键词: Amlexanox  Bone marrow mesenchymal stem cell  Osteogenesis  Osteoporosis
基金项目:国家自然科学基金(81472133)、湖北省技术创新专项重大项目(2016ACA149)、同济医院科研基金(2017B017)
作者单位E-mail
张勇 430030 武汉华中科技大学同济医学院附属同济医院骨科  
关邯峰 430030 武汉华中科技大学同济医学院附属同济医院骨科  
方忠 430030 武汉华中科技大学同济医学院附属同济医院骨科  
吴巍 430030 武汉华中科技大学同济医学院附属同济医院骨科  
熊伟 430030 武汉华中科技大学同济医学院附属同济医院骨科  
李锋 430030 武汉华中科技大学同济医学院附属同济医院骨科 lifengmd@hust.edu.cn 
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中文摘要:
      目的 探讨氨来呫诺调控大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSCs)成骨分化的作用,为其在抗骨质疏松治疗等方面的应用提供理论依据。方法 体外分离、培养SD大鼠BMSCs。①通过CCK8法检测不同浓度氨来呫诺对BMSCs增殖的影响;②设置对照组和氨来呫诺(25 μmol/L)处理组,利用成骨诱导培养基进行诱导,诱导后第10天进行碱性磷酸酶(alkaline phosphatase, ALP)染色检测;诱导后第21天进行茜素红染色;③实时荧光定量PCR检测诱导后第3、7、14天成骨分化相关基因ALP、Runt相关转录因子2(runt-related transcription factor 2, RUNX2)、骨桥蛋白(osteopontin, OPN)的表达水平。结果 ①CCK8检测结果显示25 μmol/L氨来呫诺对BMSCs增殖没有明显影响,50 μmol/L氨来呫诺显著抑制BMSCs的增殖,故在后续实验中均采用25 μmol/L氨来呫诺作为效应浓度。②干预后第10天,氨来呫诺处理组较对照组的ALP染色阳性面积明显增多;干预后第21天,氨来呫诺处理组较对照组的钙结节形成密集,钙结节数目明显增多。③干预后第7天,氨来呫诺处理组较对照组中ALP的表达显著提高;第14天时,氨来呫诺处理组较对照组中RUNX2、OPN的表达显著提高。结论 氨来呫诺可能通过促进BMSCs成骨分化相关基因表达,进而促进BMSCs细胞中ALP的表达和钙结节形成,促进BMSCs成骨分化。
英文摘要:
      Objective To investigate the effect of amlexanox on osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and provide theoretical basis for anti-osteoporosis treatment. Methods The SD rat BMSCs were isolated and cultured in vitro. The effect of different concentrations of amlexanox on the proliferation of BMSCs was detected by CCK8 assay. The cells were divided into control group and amlexanox (25 μmol/L) treatment group, and they were induced by osteogenic induction medium. Alkaline phosphatase (ALP) staining test was performed on the 10th day and alizarin red staining on the 21st day. The expression levels of ALP, runt-related transcription factor 2 (RUNX2) and osteopontin (OPN) mRNA on the day 3, 7 and 14 were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Results CCK8 test results showed that 25 μmol/L amlexanox had no significant effect on the proliferation of BMSCs, but 50 μmol/L amlexanox did, so the 25 μmol/L amlexanox was used for the effect concentration. The positive staining area in amlexanox group was significantly increased at 10th day after treatment and the formation of calcium nodules in amlexanox group was increased at the 21st day compared to the control group. The mRNA expression levels of ALP were significantly promoted at the 7th day, and the expression levels of RUNX2 and OPN were simultaneously promoted at the 14th day. Conclusion Amlexanox could promote the osteogenic differentiation of BMSCs by promoting the expression of osteogenic differentiation related genes of BMSCs.
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