CAJ | 학술논문

背景:缺血缺氧的心肌微环境导致植入的细胞存活率低.目的:观察沉默 caspase-3基因对大鼠骨髓间充质干细胞增殖和体外缺血缺氧环境下凋亡的影响.方法:构建靶向 caspase-3的 shRNA 重组慢病毒并转染骨髓间充质干细胞为转基因组,以正常细胞组和空载体组做对照,采用 MTS 法检测各组细胞增殖情况.建立缺血缺氧模型,real-time PCR 和免疫组织化学分别检测缺血缺氧环境各组细胞的 caspase-3 mRNA 和蛋白表达水平,应用流式细胞术检测不同缺血缺氧时间点(0,6,12,24,48 h)各组细胞的凋亡率.结果与结论:重组慢病毒成功转染骨髓间充质干细胞,且细胞增殖活性升高(P <0.05).缺血缺氧环境下,转基因组细胞 caspase-3在 mRNA 和蛋白表达水平相比对照组下降(P <0.05).沉默 caspase-3能显著降低骨髓间充质干细胞的凋亡率(P <0.05),且随着缺血缺氧时间的延长凋亡率缓慢升高.结果提示,沉默 caspase-3能加快骨髓间充质干细胞的生长速度和提高在体外缺血缺氧环境下的抗凋亡能力.
배경:결혈결양적심기미배경도치식입적세포존활솔저.목적:관찰침묵 caspase-3기인대대서골수간충질간세포증식화체외결혈결양배경하조망적영향.방법:구건파향 caspase-3적 shRNA 중조만병독병전염골수간충질간세포위전기인조,이정상세포조화공재체조주대조,채용 MTS 법검측각조세포증식정황.건립결혈결양모형,real-time PCR 화면역조직화학분별검측결혈결양배경각조세포적 caspase-3 mRNA 화단백표체수평,응용류식세포술검측불동결혈결양시간점(0,6,12,24,48 h)각조세포적조망솔.결과여결론:중조만병독성공전염골수간충질간세포,차세포증식활성승고(P <0.05).결혈결양배경하,전기인조세포 caspase-3재 mRNA 화단백표체수평상비대조조하강(P <0.05).침묵 caspase-3능현저강저골수간충질간세포적조망솔(P <0.05),차수착결혈결양시간적연장조망솔완만승고.결과제시,침묵 caspase-3능가쾌골수간충질간세포적생장속도화제고재체외결혈결양배경하적항조망능력.
@@@@BACKGROUND: Ischemia and hypoxia myocardial microenvironment leads to poor survival of transplanted cel s. OBJECTIVE: To investigate the effects of silencing caspase-3 gene on proliferation and apoptosis of rat bone marrow mesenchymal stem cel s under ischemia and hypoxia in vitro. METHODS: Lentiviral short hairpin RNA interference vector targeting caspase-3 was constructed and transfected into mesenchymal stem cel s as the transgene group. The normal cel group and the empty vector group were as controls. MTS assay was applied to examine the proliferation of cel s in each group. The ischemia and hypoxia model was established. The expressions of caspase-3 mRNA and protein were detected by real-time PCR and immunohistochemistry. The apoptotic rates of the cel s at different time points (0, 6, 12, 24 and 48 hours) in each group were evaluated by flow cytometry. RESULTS AND CONCLUSION: Recombinant lentivirus was transfected into mesenchymal stem cel s successful y and the proliferation activity of the cel s was increased (P < 0.05). Compared with control groups, the levels of caspase-3 mRNA and protein in the transgene group were decreased (P < 0.05) under ischemia and hypoxia. Silencing caspase-3 could reduce the apoptotic rate of mesenchymal stem cel s (P < 0.05), and the apoptotic rate was increased slowly as ischemia and hypoxia time prolonging. Silencing caspase-3 can increase the growth speed and the anti-apoptosis ability of mesenchymal stem cel s under ischemia and hypoxia in vitro.