中山大学学报（医学科学版）2012，Vol.33Issue(1)：1-7,7.
基因干扰Sirtuin3后对缺氧预处理的影响
Effect of Sirtuin3 on Hypoxic Preconditioning with Its Gene Knocked Down
摘要
Abstract
[ Objective] To investigate the effect of Sirtuin3 (SIRT3) on hypoxic preconditioning (Hyp) with its gene knocked down. [Methods] Empty vectors (pGCSIL-GFP) were used to select the suitable MOI. pGCSIL-PUR-SIRT3 -vectors were used in formal experiments. Cells transfected with empty vectors or SIRT3-RNAi-LV were divided into control, hypoxic preconditioning + Oxygen-glucose deprivation (Hyp+OGD) and OGD respectively. MTT assay was used for testing the cells viability. The ATP level was evaluated using ATP Fluorometric Assay Kit. The expression of SIRT3, Peroxisome proliferator-activated receptor-7 coactivator-lα (PGC-lα) and Manganese superoxide dismutase (MnSOD) were assessed at the protein level by Western blot. [Results] After OGD, MTT values decreased more in SIRT3-/- compared with Vector (0.430 ± 0.009 vs 0.562 ±0.140, P < 0.01), but were increased in Hyp + OGD respectively (P < 0.05) , resulting in no significant difference between SIRT3-/- and Vector in Hyp+OGD(P = 0.311 > 0.05). After OGD, The ATP levels decreased more in SIRT3-/- compared with Vector (0.334 ± 6.006 vs 0.453 ± 0.015,P<0.01),but were increased in Hyp+OGD respectively (P < 0.05), resulting in no significant difference between SIRT3-/- and Vector in Hyp+ OGD (P = 0.866 > 0.05). The expression of PGC-lα and MnSOD were declined significantly in the three groups of SIRT3-/-compared with that in Vector respectively (P<0.05), but were both increased in Hyp+OGD and OGD compared with control (P < 0.05), and were increased more in Hyp+OGD compared with OGD (P < 0.05). [Conclusion] The expression of PGC-la and MnSOD were decreased with SIRT3 gene knocked down and SIRT3 was one of the ways that hypoxic preconditioning or hypoxia up regulated the expression of MnSOD through PGC-la pathway.关键词
缺氧预处理/Sirtuin3/过氧化物酶体增生激活受体-γ的共刺激因子-1α/锰超氧化物歧化酶/siRNA干扰Key words
hypoxic preconditioning/ Sirtuin3/ peroxisome proliferator-activated receptor-7 coactivator-lot/ manganese superoxide dismutase/ RNA interference分类
医药卫生引用本文复制引用
李超英,胡俊,赵嘉,王莹,陶玉倩,李玲..基因干扰Sirtuin3后对缺氧预处理的影响[J].中山大学学报（医学科学版）,2012,33(1):1-7,7.基金项目
国家自然科学基金(81071069) （81071069）
广东省科技计划项目(2010B080701008) （2010B080701008）
广东省科技计划项目(2008B080703029) （2008B080703029）
