CAJ | 학술논문

目的观察骨形态发生蛋白-2(BMP-2)、胰岛素样生长因子-1(IGF-1)在高糖环境下能否转染大鼠骨髓间充质干细胞(BMSC),转染成功后目的基因表达.方法腺病毒载体介导大鼠BMP-2和IGF-1基因高糖环境下转染Wistar大鼠BMSC,应用逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附试验(ELISA)定量检测基因复制及表达.结果　高糖环境下BMP-2、IGF-1基因各自及共同转染BMSC后,BMSC成活,BMP-2基因表达的产物峰值出现在转染后72 h,BMP-2组灰度值为(1.86±0.06),产物浓度为(58.60±1.61)μg/L,BMP-2+IGF-1组灰度值为(2.37±0.34),产物浓度为(67.61±1.91)μg/L,两组差异均有统计学意义(P＜0.05);IGF-1基因表达的产物峰值出现在转染后48 h,IGF-1组灰度值为(1.48±0.03),产物浓度为(65.17±3.64)μg/L,BMP-2+IGF-1组灰度值为(1.93±0.17),产物浓度为(73.94±4.30)μg/L,两组差异均有统计学意义(P＜0.05).结论　高糖环境下BMP-2、IGF-1可转染BMSC,且转染后基因表达产物量显著增加.BMP-2、IGF-1基因在基因表达上有协同作用.
목적 관찰골형태발생단백-2(BMP-2)、이도소양생장인자-1(IGF-1)재고당배경하능부전염대서골수간충질간세포(BMSC),전염성공후목적 기인표체.방법 선병독재체개도대서BMP-2화IGF-1기인고당배경하전염Wistar대서BMSC,응용역전록-취합매련반응(RT-PCR)화매련면역흡부시험(ELISA)정량검측기인복제급표체.결과　고당배경하BMP-2、IGF-1기인각자급공동전염BMSC후,BMSC성활,BMP-2기인표체적산물봉치출현재전염후72 h,BMP-2조회도치위(1.86±0.06),산물농도위(58.60±1.61)μg/L,BMP-2+IGF-1조회도치위(2.37±0.34),산물농도위(67.61±1.91)μg/L,량조차이균유통계학의의(P＜0.05);IGF-1기인표체적산물봉치출현재전염후48 h,IGF-1조회도치위(1.48±0.03),산물농도위(65.17±3.64)μg/L,BMP-2+IGF-1조회도치위(1.93±0.17),산물농도위(73.94±4.30)μg/L,량조차이균유통계학의의(P＜0.05).결론　고당배경하BMP-2、IGF-1가전염BMSC,차전염후기인표체산물량현저증가.BMP-2、IGF-1기인재기인표체상유협동작용.
Objective To investigate whether bone morphogenetic protein-2 (BMP-2) and insulin-like growth factor-1 ( IGF-1 ) in high glucose can be transfected into Wistar rat mesenchymal stem cells,and the gene expression after transfection. Methods Adenovirus-mediated BMP-2 and IGF-1 genes were transfected into the Wistar rat mesenchymal stem cells. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the gene replication and expression. Results After the transfection of adenovirus-mediated BMP-2 and(or) IGF-1 gene into the cells in high glucose, all of bone marrow stromal cells survived, bone marrow stromal cells could express the target gene product with the peak of BMP-2 gene product expression at in 72nd h after transfection. The grey degree and gene product concentration of BMP-2 in BMP-2 group were ( 1.86 ± 0. 06) and (58.60 ±1.61) μg/L, and those in BMP-2 ± IGF-1 group (2.37 ±0.34) and (67.61 ±1.91) μg/L,respectively (all P ＜ 0. 05 ). The peak of IGF-1 gene expression occurred at 48th h after transfection. The grey degree and gene product concentration of IGF-1 in IGF-1 group were ( 1.48 ± 0. 03 ) and ( 65.17 ±3.64) μg/L, and those in BMP-2 + IGF-1 group were (1.93 ±0. 17) and (73. 94 ±4. 30) μg/L, respectively ( all P ＜0. 05). Conclusion Adenovirus-mediated BMP-2 and IGF-1 gene could be transfected the Wistar rat bone marrow stromal cells in high glucose. The gene products after transfection were increased significantly. There was a synergistic effect on gene expression between BMP-2 and IGF-1.