Intracellular ceramide accumulation associated with apoptosis and changes in energy metabolism in primary cultured myocytes exposed to hypoxia and re-oxygenation

细胞内神经酰胺积累与缺氧和复氧原代培养心肌细胞凋亡和能量代谢变化相关

• 批准号: 17591461
• 负责人: TAKEUCHI Koh
• 金额: $ 2.05万
• 依托单位: The University of Tokyo (2006)National Research Institute for Child Health and Development (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591461/
• 关键词: Ischemic injury; Re-oxygenation injury; glucose metabolism; fatty acid metabolism; ceramide; apoptosis; 心筋細胞; 虚血-再酸素化

This study has been designed to see the intracellular event in myocytes during hypoxia-reoxygenation process. Cultured myocytes were prepared and exposed to hypoxia for 90 minutes followed by 60 minutes of re-oxygenation. Glucose extraction, lactate production, apoptotic cell death, extracellular regulated protein kinase (ERKs) phophorylation were mainly measured. Specific sphyngomyelinase inhibitor, D-609 was also tested to see if there was protective effects on myocytes exposed to hypoxia followed by re-oxygenation insult. During hypoxia lactate production was significantly higher in control than D-609 group, whereas glucose uptake was significantly reduced in control group compared to D609 group. Apoptotic cell death was clearly decreased with D 609 treatment with significantly reduced phosphorylation of ERKs. However one of the marker for cell necrosis, fatty acid binding protein FABP was not different between the group, suggesting that clearly detected apoptosis is different from necrotic cell death. Ceramide Protein synthesis measured by Western immunoblotting was significantly lower in D 699 group than control. This may imply that D609 is contributed to a reduction of ceramide accumulation in hypoxia-reoxygenation exposed myocytes. Based on our findings, we conclude that accumulation of ceramide might be associated with impairement of glucose metabolism, possibly fatty acid metabolism as well as significantly increased apoptotic cell death. Suppression of ceramide accumulation in myocytes might be contributed to reduced apoptotic cell death and extracellular regulated protein kinase phosphorylation as well as improved glucose metabolism.

这项研究旨在在缺氧 - 抗氧化过程中查看肌细胞的细胞内事件。制备培养的肌细胞并暴露于缺氧90分钟，然后重新氧化60分钟。主要测量葡萄糖提取，乳酸产生，凋亡细胞死亡，细胞外调节的蛋白激酶（ERKS）噬菌体。还测试了特异性杂菌素酶抑制剂D-609，以查看是否对暴露于缺氧的心肌细胞有保护作用，然后再重新氧气侮辱。在对照组中，乳酸在缺氧期间的产生显着高于D-609组，而与D609组相比，对照组的葡萄糖摄取显着降低。 D 609治疗明显降低了凋亡细胞死亡，而ERK的磷酸化显着降低。然而，脂肪酸结合蛋白FABP的细胞坏死标记之一是该组之间的差异，这表明清晰检测到的凋亡与坏死细胞死亡不同。 D 699组中通过西部免疫印迹测得的神经酰胺蛋白合成明显低于对照。这可能意味着D609有助于降低神经酰胺在低氧抗氧化中暴露的心肌细胞的积累。根据我们的发现，我们得出结论，神经酰胺的积累可能与葡萄糖代谢，可能是脂肪酸代谢以及凋亡细胞死亡的显着增加有关。肌细胞中神经酰胺积累的抑制可能导致凋亡细胞死亡减少和细胞外调节的蛋白激酶磷酸化以及改善的葡萄糖代谢。

项目成果

Surgical Outcome of Heterotaxy Syndrome in a Single Institution

单一机构异位综合征的手术结果

• 发表时间: 2006
• 期刊: Asian Cardiovasc Thorac Ann 14
• 作者: Koh Takeuchi;et al.
• 通讯作者: et al.

Fate of Equine Pericardial Roll Conduit for Rastelli Operation during Long-term Follow-up

长期随访期间用于 Rastelli 手术的马心包滚动导管的命运

• 发表时间: 2007
• 期刊: Congenital Heart Diseases 2
• 作者: Koh Takeuchi;et al.
• 通讯作者: et al.

Surgical Outcome of Heterotaxy Syndrome in a Single Institution.

单一机构异位综合征的手术结果。

• 发表时间: 2006
• 期刊: Asian Cardiovasc Thorac Ann 14
• 作者: Koh Takeuchi;et al.
• 通讯作者: et al.

Evaluation of Valved Saphenous Vein Homograft as Right Ventricle-Pulmonary Artery Conduit in Modified Stage I Norwood Operation.

改良诺伍德第一阶段手术中带瓣隐静脉同种移植物作为右心室-肺动脉导管的评估。

• 发表时间: 2006
• 期刊: Interact CardioVasc Thorac Surg 5
• 作者: Koh Takeuchi;et al.
• 通讯作者: et al.