下调microRNA-34a增强对人脂肪干细胞成骨诱导分化作用的初步探索  

Primary Exploration of Promotion Effect of microRNA-34a Knockdown on Human Adipose-Derived Stem Cells in Osteoinduction Differentiation

作　　者：李鹏飞 吴佳君 李余 张静 吴慧玲 LIPeng-fei;WUJia-jun;LI Yu;ZHANG Jing;WU Hui-lin(The First Affiliated Hospital,Zhejiang University,Aesthetic Plastic Center,Hangzhou,Zhejiang,310000,China;Zhejiang University,School of Medicine,Hangzhou,Zhejiang,310000,China)

机构地区：[1]浙江大学附属第一医院整形美容中心,浙江杭州310000 [2]浙江大学医学院,浙江杭州310000

出　　处：《现代生物医学进展》2019年第3期406-410,共5页Progress in Modern Biomedicine

基　　金：传染病诊治国家重点实验室(浙江大学)国家自然科学基金面上项目(81671926)

摘　　要：目的:颅颌面临界骨缺损是整复外科常见疾患,常常给患者的身心带来障碍和压力。为解决骨形成不足这一难题,我们拟通过表观遗传修饰的手段,调节microRNA-34a在人脂肪干细胞中的表达,来探讨microRNA-34a对人脂肪干细胞成骨诱导分化的影响。方法:分离培养原代人脂肪干细胞,以慢病毒为转染载体,分三组对其进行上调、下调microRNA-34a的表达及阴性对照。然后对其进行成骨诱导培养,于诱导的第7天行碱性磷酸酶(ALP)染色,第14天行茜素红(Alizarin Red)染色,定性比较转染后人脂肪干细胞成骨诱导分化的效果。结果:成功分离和培养原代人脂肪干细胞,并以慢病毒高效率转染调控microRNA-34a的表达。对比碱性磷酸酶和茜素红染色可见下调microRNA-34a组成骨效率最高,其次是阴性对照组,上调组效率最低。结论:1.人脂肪干细胞通过慢病毒转染获得高效率的表观遗传修饰;2.下调microRNA-34a的表达对人脂肪干细胞成骨的诱导分化有一定的促进作用。Objective: Cranial critical-sized bone defect is commonly seen in plastic and reconstructive surgery causing dysfunctions and pressure on patients both physically and psychologically. In order to solve the difficulty, we regulated the expression of microRNA-34 a in human adipose-derived stem cells(hADSCs) by epigenetic modification to evaluate the effect of microRNA-34 a on hADSCs in osteoinduction differentiation. Methods: Extract and culture primary h ADSCs. Transfect h ADSCs with lentivirus respectively in three groups: microRNA-34 a up-regulation, knockdown and negative control. And then induce them to differentiate into osteoblasts.The osteogenic efficiency was qualitatively detected by ALP staining at day 7 and alizarin red staining at day 14 after osteoinduction.Results: hADSCs were successfully extracted and cultured;microRNA-34 a expression was regulated by means of high efficient transfection of lentivirus;staining results showed that microRNA-34 a knockdown group was most efficient in osteoinduction differentiation, control group came as the second, up-regulation group was the least efficient. Conclusion: 1.By transfecting with lentivirus, hADSCs acquired high efficiency of epigenetic modification;2.microRNA-34 a knockdown to some extent can promote osteoinduction differentiation in hADSCs.

关 键 词：下调microRNA-34a 人脂肪干细胞 成骨诱导分化 

分 类 号：R-33[医药卫生] R329