文章摘要
张大伟,徐晋妉,卞智萍,刘 静,吴恒芳,顾春荣,陈相健,杨 笛.黄芪皂苷Ⅳ通过PKA途径减轻缺氧/复氧心肌损伤的机制研究[J].南京医科大学学报,2012,(1):30~34
黄芪皂苷Ⅳ通过PKA途径减轻缺氧/复氧心肌损伤的机制研究
The mechanism of astragaloside IV alleviation of hypoxia/reoxygenation induced cultured cardiomyocyte injury via PKA pathway
投稿时间:2011-09-28  
DOI:10.7655
中文关键词: 黄芪皂苷Ⅳ  心肌保护  肌浆网钙ATP酶  PKA  受磷蛋白
英文关键词: astragaloside IV  cardioprotection  sarcoplasmic reticulum calcium ATPase  PKA  phosphorylated phospholamban
基金项目:国家自然科学基金项目(30772781,30770894)
作者单位
张大伟 南京医科大学第一附属医院分子心脏病实验室/心内科,江苏 南京 210029 
徐晋妉 南京医科大学第一附属医院分子心脏病实验室,江苏 南京 210029 
卞智萍 南京医科大学第一附属医院分子心脏病实验室,江苏 南京 210029 
刘 静 南京医科大学第一附属医院老年科,江苏 南京 210029 
吴恒芳 南京医科大学第一附属医院分子心脏病实验室,江苏 南京 210029 
顾春荣 南京医科大学第一附属医院分子心脏病实验室,江苏 南京 210029 
陈相健 南京医科大学第一附属医院分子心脏病实验室,江苏 南京 210029 
杨 笛 南京医科大学第一附属医院心内科,江苏 南京 210029 
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中文摘要:
      目的:研究黄芪皂苷IV(astragaloside IV,As-IV)减轻缺氧/复氧所致心肌细胞损伤的机制?方法:胰酶多次消化法培养新生SD大鼠原代心肌细胞,建立缺氧/复氧模型,测定培养心肌细胞上清心肌损伤标志物肌酸激酶同工酶(creatine kinase,CK-MB)含量,检测心肌细胞肌浆网钙ATP酶(sarcoplasmic reticulum Ca2+-ATPase,SERCA2a)活性?Real-Time PCR法测定PKA催化亚单位α(PKA C subunit α,PKA- Cα)基因的表达水平以及Western blot检测第16位丝氨酸磷酸化受磷蛋白(Ser16 phosphorylated phospholamban,Ser16-PLN)的表达水平,同时以As-IV(30 ?滋mol/L)干预,观察其作用?结果:与正常组相比,缺氧/复氧(hypoxia/reoxygenation,H/R)引起心肌细胞CK-MB释放增加,SERCA2a活性降低约35%?PKA- Cα基因表达下调28%以及Ser16-PLN表达下调51%,P值均﹤0.05,As-IV干预则可逆转上述变化,基本恢复至正常水平?结论:黄芪皂苷Ⅳ抑制缺氧/复氧所致心肌细胞损伤的机制可能是通过上调PKA-Cα基因表达,提高受磷蛋白(phosphorylated phospholamban,PLN)16位丝氨酸的磷酸化水平,解除PLN对SERCA2a的抑制,从而增强SERCA2a的功能?
英文摘要:
      Objective: To investigate the mechanism of astragaloside IV(As-IV) on alleviation of hypoxia/reoxygenation induced myocardial injury. Methods: Cultured cardiomyocytes from neonatal SD rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation(H/R). Meanwhile,As-IV (30 ?滋mol/L) was treated in H/R cardiomyocytes. Myocytes injury was determined by the release of creatine kinase (CK-MB) in supernatant. Myocardial SERCA2a activity was measured,PKA C subunit α (PKA-Cα) gene expression and Ser16 phosphorylated phospholamban(Ser16-PLN) protein expression level were detected by real-time PCR and Western blot respectively. Results: Cultured cardiomyocytes exposed to hypoxia/reoxygenation presented statistically higher CK-MB in supernatant,decreased myocardial SERCA2a activity,reduced PKA-Cα gene and Ser16-PLN protein expressions(P < 0.05). But As-IV treatment significantly prevented the alterations in H/R cardiomyocytes mentioned above. Conclusion: These results suggest that the cardioprotective effects of As-IV may be associated with upregulation of PKA-Cα gene and Ser16-PLN protein expressions,thus restoring the SERCA2a function in hypoxia/reoxygenation injury.
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