CAJ | 학술논문

背景：骨形态发生蛋白7具有促进骨髓间充质干细胞分裂增殖的作用，在诱导新骨形成方面有重要的研究意义。目的：观察携带人骨形态发生蛋白7慢病毒载体转染后骨髓间充质干细胞骨形态发生蛋白7蛋白及mRNA的表达。方法：提取并培养大鼠骨髓间充质干细胞，通过形态学和细胞表面标记进行鉴定；构建携带人骨形态发生蛋白7的慢病毒载体，转染大鼠骨髓间充质干细胞，提取RNA后采用RT-PCR的方法检测骨形态发生蛋白7 mRNA水平相对表达量的变化，Western blotting检测骨形态发生蛋白7蛋白水平相对表达量的变化。结果与结论：成功构建了携带骨形态发生蛋白7基因的慢病毒载体PLV-sfGFP(2A)-BMP7，重组载体质粒成功转染至大鼠骨髓间充质干细胞中，转染组的骨形态发生蛋白7 mRNA和蛋白表达高于空载体组和空白组，差异有显著性意义(P <0.05)，说明外源性骨形态发生蛋白7基因被有效整合到骨髓间充质干细胞并进行表达。
배경：골형태발생단백7구유촉진골수간충질간세포분렬증식적작용，재유도신골형성방면유중요적연구의의。목적：관찰휴대인골형태발생단백7만병독재체전염후골수간충질간세포골형태발생단백7단백급mRNA적표체。방법：제취병배양대서골수간충질간세포，통과형태학화세포표면표기진행감정；구건휴대인골형태발생단백7적만병독재체，전염대서골수간충질간세포，제취RNA후채용RT-PCR적방법검측골형태발생단백7 mRNA수평상대표체량적변화，Western blotting검측골형태발생단백7단백수평상대표체량적변화。결과여결론：성공구건료휴대골형태발생단백7기인적만병독재체PLV-sfGFP(2A)-BMP7，중조재체질립성공전염지대서골수간충질간세포중，전염조적골형태발생단백7 mRNA화단백표체고우공재체조화공백조，차이유현저성의의(P <0.05)，설명외원성골형태발생단백7기인피유효정합도골수간충질간세포병진행표체。
BACKGROUND:Bone morphogenetic protein-7 can promote proliferation of bone marrow mesenchymal stem cels and has important significance in terms of inducing new bone formation. OBJECTIVE:To observe the protein and mRNA expression of bone morphogenetic protein-7 in bone marrow mesenchymal stem cels transfected with bone morphogenetic protein-7. METHODS:Rat bone marrow mesenchymal stem cels were isolated and cultured, and then identified by the morphology and cel surface markers. Lentiviral vectors carrying bone morphogenetic protein-7 were constructed to transfect bone marrow mesenchymal stem cels. After total RNA extraction, the protein and mRNA expression of bone morphogenetic protein-7 was detected by RT-PCR and western blot methods. RESULTS AND CONCLUSION:PLV-sfGFP(2A)-BMP7 recombinant plasmids were successfuly constructed and transferred into rat bone marrow mesenchymal stem cels. The mRNA and protein expression of bone morphogenetic protein-7 in the transfection group was significantly higher than that in the blank vector and blank groups (P < 0.05), indicating exogenous bone morphogenetic protein-7 gene is effectively integrated into the bone marrow mesenchymal stem cels.