张聘,张林香,赵建宁,张雷,等.抑制microRNA-202-3p表达促进滑膜间充质干细胞软骨分化[J].,2018,5(5):21-25.
抑制microRNA-202-3p表达促进滑膜间充质干细胞软骨分化
Downregulation of MicroRNA-202-3p enhances chondrogenic differentiation of human synovium-derived mesenchymal stem cells
投稿时间:2018-01-05  最后修改时间:2018-01-20
DOI:10.12095/j.issn.2095-6894.2018.05.005
中文关键词:  滑膜间充质干细胞;细胞分化;软骨细胞;MicroRNA
英文关键词:synovial mesenchymal stem cells; proliferation; chondrocyte differentiation; Has?miR?202?3p
基金项目:国家自然科学基金青年科学基金项目(81702170)、江苏省自然科学基金青年基金项目(BK20170624)、中国博士后科学基金特别资助项目(2017T00826)
作者单位E-mail
张聘 南京医科大学金陵临床医学院 1430640566@qq.com 
张林香 解放军南京总医院麻醉科  
赵建宁 解放军南京总医院骨科  
张雷 解放军南京总医院骨科 E-mail: ra_eagle@hotmail.com 
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中文摘要:
      【摘 要】目的:探究microRNA?202?3p(miR?202?3p)对滑膜间充质干细胞(SMSCs)软骨分化能力的影响。方法:从骨关节炎   (OA)患者手术切除的滑膜组织标本中分离培养SMSCs,分别以Has?miR?202?3p inhibitor 和Has?miR?202?3p inhibitor negative con?   trol(NC)转染第3 代SMSCs,分别设为实验组和对照组,培养7 d 内,检测两组细胞增殖曲线;培养21 d 后,组织学观察两组成软   骨能力;qRT?PCR 检测两组成软骨分化后细胞的软骨特异性基因表达。结果:滑膜间充质干细胞从第3 天开始进入快速增殖期,   CCK?8 实验和亚甲蓝染色显示实验组细胞增殖数量明显多于对照组;成软骨诱导21 d 后,实验组软骨特异性基因II 型胶原蛋   白、蛋白多糖、Sox9 基因表达水平显著高于对照组(P<0.05)。结论:抑制microRNA?202?3p 表达可促进滑膜间充质干细胞增殖和   向软骨细胞分化。
英文摘要:
      【Abstract】Objective: To study the effect of microRNA?202?3p(miR?202?3p) on the chondrogenesis of synovium?derived mesenchymal   stem cells(SMSCs). Methods: SMSCs were separated and purified from the synovium of osteoarthritis (OA) patients. Passage 3 SMSCs   were transfected with miR?202?3p inhibitor(experimental) and miR?202?3p inhibitor negative control (control). Cell viability was esti?   mated at 1, 2, 3, 4, 5, 6 and 7 days. The passage 3 cells were cultured in the chondrogenic medium for 21 days. Then the chondrogene?   sis potential of cells was examined by toluidine blue staining. The chondrogenic?specific marker genes Type II collagen(Col2), Aggrecan   and Sox9 were determined by Real?time PCR. Results: The cell growth curve exhibited rapid proliferation after 3 days. CCK?8 assay and   toluidine blue staining showed suppression of miR?202?3p significantly promoted proliferation in synovium?derived mesenchymal stem cells.   Furthermore, we detected the higher expression of chondrogenic specific marker genes, such as Col2, Aggrecan and Sox9, in the experi?   mental group than the control group at 21 days after chondrogenic induction (P<0.05). Conclusion: Downregulation of miRNA?202?3p   could enhance proliferation and chondrogenic differentiation of SMSCs.
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