Ad-GLUT3转染对缺氧再灌注损伤PC12细胞的影响
Adenovirus-mediated glucose transporter 3 infection in PC12 cells of rats after hypoxiareperfusion injury
摘要目的 探讨腺病毒介导的葡萄糖转运体3(Ad-GLUT3)对缺氧再灌注损伤的PC12细胞的影响.方法 在构建大鼠PC12细胞缺氧再灌注损伤模型的基础上,使用Ad-GLUT3转染PC12细胞,并以Ad-GFP转染和空白质粒转染为对照.流式细胞仪和TUNEL法进行细胞凋亡检测,同位素放射法分析葡萄糖代谢率情况,Western blotting分析细胞凋亡相关蛋白核因子-KB(NF-KB)的表达.结果 流式细胞仪结果显示,Ad-GLUT3组凋亡率(4.7%±1.75%)较Ad-GFP组(14.25%±3.46%)和空白对照组(13.1%±2.98%)明显下降,差异有统计学意义(P<0.05);同时TUNEL 法结果显示,Ad-GLUT3组凋亡率(7.7%±3.31%)较Ad-GFP组(17.1%±3.10%)和空白对照组(15.2%±2.70%)明显下降,差异有统计学意义(P<0.05).Ad-GLUT3组葡萄糖摄取率(603%±13.1%)较Ad-GFP 组(300%±8.77%)和空白对照组(319%±11.2%)明显提高,差异有统计学意义(P<0.05).Ad-GLUT3组NF-kB被激活的程度(0.92±0.17)较Ad-GFP组(1.02±0.24)和空白对照组(1.06±0.21)明显下降,差异有统计学意义(P<0.05).结论 Ad-GLUT3转染缺氧再灌注损伤的PC12细胞可以有效提高细胞的能量代谢,抑制细胞凋亡.
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abstracts[Objective]To investigate the effect of adenovirus-mediated glucose transporter 3 (Ad-GLUT3) infection on PC 12 cells of rats after hypoxia-reperfusion injury.[Methods]PC 12 cells treated with hypoxia and re--oxygenation were prepared;after PC12 cells being infected by Ad-GLUT3 (experimental group),adenovirus encoding green fluorescent protein (Ad-GFP group) and blank plasmid (control group),respectively,flow cytometry (FC) and TUNEL assay were used to analyze the apoptosis of PC12 cells;the glucose uptake rate was detected by isotopic-scintiscan,and the expression of nuclear factor (NF-kB) was detected by Western blotting.[Results] FC indicated that the apoptotic rate in the experimental group (4.7%±1.75%) was significantly lower as compared with that in the Ad-GFP group (14.25%±3.46%) and control group (13.1%±2.98%,P<0.05);TUNEL assay showed that the apoptotic rate in the experimental group (7.7%±3.31%) was significantly lower as compared with that in the Ad-GFP group (17.1%±3.10%) and control group (15.2%±2.70%,P<0.05).And the glucose uptake rate in the experimental group (603%±13.1%) was obviously higher as compared with that in the Ad-GFP group (300%±8.77%) and control group (319%±11.2%,P<0.05);and the NF-k<B level in the experimental group (0.92±0.17) was significantly lower as compared with that in the Ad-GFP group (1.02±0.24) and control group (1.06±0.21,P<0.05).[Conclusion] After being infected by Ad-GLUT3,the PC12 cells treated with hypoxia and re-oxygenation enjoy elevated glucose uptake rate and inhibited apoptotic rate.
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