CAJ | 학술논문

目的探讨肿瘤坏死因子样弱凋亡因子(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)基因转染骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)后对其增殖功能的影响.方法全骨髓贴壁法筛选培养幼年大鼠BMSCs,用腺病毒载体AdSCMV-TWEAK转染TWEAK基因,通过逆转录一聚合酶链式扩增实验(RT-PCR)观察大鼠BMSCs细胞转染后TWEAK蛋白的表达情况.实验分成3组,即转染Ad5CMV-TWEAK的实验组,转染Ad5CMV-EGFP的对照组及未转染任何基因的空白对照组.用MTr法比较各组BMSCs细胞的增殖变化情况.结果全骨髓贴壁培养法能有效分离纯化大鼠BMSCs,RT-PCR可证明AdSCMV-TWEAK成功的转染BMSCs 细胞,转染后在BMSCs产生较高的表达.MTY值经χ~2检验,TWEAK组与无干预组比较差异显著(P<0.05),而EGFP组与无干预组无显著差异(P>0.05).结论 TWEAK转染有刺激BMSCs的增殖作用,如进一步证明其促分化作用,其在骨修复材料的改进以及种植体涂层修饰方面将有广阔的应用前景.
목적 탐토종류배사인자양약조망인자(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)기인전염골수간충질간세포(bone mesenchymal stem cells,BMSCs)후대기증식공능적영향.방법 전골수첩벽법사선배양유년대서BMSCs,용선병독재체AdSCMV-TWEAK전염TWEAK기인,통과역전록일취합매련식확증실험(RT-PCR)관찰대서BMSCs세포전염후TWEAK단백적표체정황.실험분성3조,즉전염Ad5CMV-TWEAK적실험조,전염Ad5CMV-EGFP적대조조급미전염임하기인적공백대조조.용MTr법비교각조BMSCs세포적증식변화정황.결과 전골수첩벽배양법능유효분리순화대서BMSCs,RT-PCR가증명AdSCMV-TWEAK성공적전염BMSCs 세포,전염후재BMSCs산생교고적표체.MTY치경χ~2검험,TWEAK조여무간예조비교차이현저(P<0.05),이EGFP조여무간예조무현저차이(P>0.05).결론 TWEAK전염유자격BMSCs적증식작용,여진일보증명기촉분화작용,기재골수복재료적개진이급충식체도층수식방면장유엄활적응용전경.
Objective To approach the effect of tumor necrosis factor-like weak inducer of apoptosis (TWEAK)gene on the proliferation of rat bone mesenchymal stem cell(BMSC).Methods The rat BMSCs were isolated and purified from bone marrow of young rats by adherent cuhure.TWEAK genes were transfected by Ad5 CMV-TWEAK.and RT-PCR was used to observe the expression of TWEAK.Then the cells were divided into 3 groups,transfected Ad5 CMV-TWEAK group,transfected Ad5CMVEGFP group and untransfected group,and the MTT assay was used to test the proliferation effect.Results The purified BMSCs were obtained by adherent culture.RT-PCR results showed high TWEAK expression after Ad5CMV-TWEAK transfected.The MTT assay results demonstrated significant difference between Ad5 CMV-TWEAK group and untransfected group(P<0.05)but no difierencebetween the transfected Ad5 CMV-EGFP group and untransfected group(P>0.05).Conclusion Transfecting TWEAK gene can stimulate BMSCs proliferation.the next research will test if it can promote the differentiation of the cells to show applicationr prospect in bone restore material and implant coating.