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靶向PUMA的siRNA对缺氧/复氧诱导大鼠心肌细胞凋亡的保护作用
引用本文:付晶晶,罗达亚,万福生. 靶向PUMA的siRNA对缺氧/复氧诱导大鼠心肌细胞凋亡的保护作用[J]. 中国生物化学与分子生物学报, 2011, 0(7)
作者姓名:付晶晶  罗达亚  万福生
作者单位:南昌大学医学院生物化学与分子生物学教研室;江西省儿童医院检验科;
基金项目:国家自然科学基金资助(No.30060029); 江西省自然科学基金资助(No.2010JZY0237)~~
摘    要:为探讨p53上调凋亡调制物(p53 up-regulated modulator of apoptosis,PUMA)在大鼠心肌细胞缺氧/复氧(hypoxia/reoxygenatio,H/R)损伤中的作用,本研究将靶向PUMA的siRNA(si-PUMA)转染大鼠心肌细胞以建立PUMA沉默表达模型,观察其对心肌细胞H/R损伤的影响.RT-PCR和Western印迹结果表明,最适转染浓度50 nmol/L si-PUMA能靶向抑制H/R损伤心肌细胞的PUMA表达;MTT法检测心肌细胞存活率及培养基乳酸脱氢酶(lactate dehydrogenase,LDH)活性测定结果发现,si-PUMA组细胞存活率较H/R 6 h模型组明显提高,培养液中LDH活性显著降低(P<0.01);分光光度法及Annexin V-FITC/PI联合染色流式细胞凋亡检测结果显示,si-PUMA组caspase-3活性较H/R 6h组明显下调,细胞凋亡率明显降低(P<0.01);RT-PCR结果提示,与H/R6 h组相比,si-PUMA组Bax及Bcl-2表达分别出现显著下调及上调(P<0.05).以上结果表明,靶向PUMA的siRN...

关 键 词:p53上调凋亡调制物  缺氧/复氧  心肌细胞  凋亡  RNA干扰  

PUMA-targeting siRNA Protects Rat Cardiomyocyte from Hypoxia/Reoxygenation-induced Apoptosis
FU Jing-Jing),),LUO Da-Ya),WAN Fu-Sheng). PUMA-targeting siRNA Protects Rat Cardiomyocyte from Hypoxia/Reoxygenation-induced Apoptosis[J]. Chinese Journal of Biochemistry and Molecular Biology, 2011, 0(7)
Authors:FU Jing-Jing)  )  LUO Da-Ya)  WAN Fu-Sheng)
Affiliation:FU Jing-Jing1),2),LUO Da-Ya1),WAN Fu-Sheng1) (1) Department of Biochemistry and Molecular Biology,Basic Medical College of Nanchang University,Nanchang 330006,China,2) Department of Clinical Laboratory,Children's Hospital of Jiangxi Province,China)
Abstract:To study the effect of p53 up-regulated modulator of apoptosis(PUMA) in hypoxia/ reoxygenation(H/R) mediated rat cardiomyocyte injury,siRNA targeted PUMA was designed and transfected into primary cultured rat cardiomyocytes in parallel with a scramble siRNA.Optimized dose of PUMA-specific siRNA was 50 nmol/L as determined by RT-PCR and Western blotting.The MTT assay and lactate dehydrogenase(LDH) activity measurement showed that the cell survival was enhanced and LDH release was reduced after the transfecti...
Keywords:p53 up-regulated modulator of apoptosis(PUMA)  hypoxia/reoxygenation  cardiomyocytes  apoptosis  RNA interference  
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