CAJ | 학술논문

背景：环氧化酶2、聚蛋白多糖酶1和胰岛素样生长因子1参与关节软骨病理损伤过程。目的：观察携带基因环氧化酶2、聚蛋白多糖酶1的shRNA干扰载体和携带胰岛素样生长因子1的过表达慢病毒载体在骨髓间充质干细胞中的表达情况。方法：应用重组慢病毒技术构建携带沉默基因环氧化酶2、聚蛋白多糖酶1、过表达基因胰岛素样生长因子1和绿色荧光蛋白基因的重组慢病毒表达载体，并用其转染体外培养的第3代人骨髓间充质干细胞(实验组)，并以无目的基因的慢病毒载体转染人骨髓间充质干细胞和未做处理的人骨髓间充质干细胞分别作为阴性对照组和空白组。结果与结论：环氧化酶2和聚蛋白多糖酶1在转染重组慢病毒的人骨髓间充质干细胞中的 mRNA 和蛋白水平有受到了明显抑制，胰岛素样生长因子1 mRNA和蛋白表达水平则明显上升。说明应用慢病毒可在人骨髓间充质干细胞成功沉默环氧化酶2和聚蛋白多糖酶1基因，同时将胰岛素样生长因子1高表达，为系统性治疗关节炎提供基因治疗的基础。
배경：배양화매2、취단백다당매1화이도소양생장인자1삼여관절연골병리손상과정。목적：관찰휴대기인배양화매2、취단백다당매1적shRNA간우재체화휴대이도소양생장인자1적과표체만병독재체재골수간충질간세포중적표체정황。방법：응용중조만병독기술구건휴대침묵기인배양화매2、취단백다당매1、과표체기인이도소양생장인자1화록색형광단백기인적중조만병독표체재체，병용기전염체외배양적제3대인골수간충질간세포(실험조)，병이무목적기인적만병독재체전염인골수간충질간세포화미주처리적인골수간충질간세포분별작위음성대조조화공백조。결과여결론：배양화매2화취단백다당매1재전염중조만병독적인골수간충질간세포중적 mRNA 화단백수평유수도료명현억제，이도소양생장인자1 mRNA화단백표체수평칙명현상승。설명응용만병독가재인골수간충질간세포성공침묵배양화매2화취단백다당매1기인，동시장이도소양생장인자1고표체，위계통성치료관절염제공기인치료적기출。
BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.