REACTIVE OXYGEN SPECIES FROM MITOCHONDRIA INDUCE CYCLOOXYGENASE-2 GENE EXPRESSION IN HUMAN MESANGIAL CELLS : POTENTIAL ROLE IN DIABETIC NEPHROPATHY

线粒体中的活性氧诱导人系膜细胞中的环加氧酶 2 基因表达：在糖尿病肾病中的潜在作用

• 批准号: 14571100
• 负责人: NISHIKAWA Takeshi
• 金额: $ 2.24万
• 依托单位: KUMAMOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571100/
• 关键词: DIABETS NELLITUS; DIABETIC NEPHROPATHY; GLOMERULAR HYPERFILTRATION; MITOCHONDRIA; OXIDATIVE STRESS; REACTIVE OXYGENSPECIES; CYCLOOXYGENASE-2; NUCLEAR FACTOR κB; プロスタグランジンE_2

Hyperglycemia increases the production of reactive oxygen species (ROS) from the mitochondrial electron transport chain in bovine endothelial cells (Nishikawa et al. 2000 ; Nature 404:787-90). Because several studies have postulated a role for prostaglandins (PGs) in the glomerular hyperfiltration seen in early diabetes, we evaluated the effect of mitochondrial ROS on expression of the inducible isoform of cyclooxygenase (COX-2) in cultured human mesangial cells (HMC). We first confirmed that incubation of HMC with 30 mM glucose significantly increased C0X-2 mRNA, but not COX-1 mRNA, compared with 5.6 mM glucose. Similarly, incubation of HMC with 30 mM glucose significantly increased mitochondrial membrane potential, intracellular ROS production, COX-2 protein expression and PGE_2 synthesis, and these events were completely suppressed by TTFA or CCCP, inhibitors of mitochondrial metabolism, or by overexpression of uncoupling protein-1 or manganese superoxide dismutase. In vivo study, it was confirmed that expression of COX-2 mRNA and protein was increased in glomeruli of diabetic mice. In addition, hyperglycemia induced activation of the COX-2 gene promoter, which was completely abrogated by mutation of two NF-κB (nuclear factor κB) binding sites in the promoter region. Our results suggest that hyperglycemia increases mitochondrial ROS production, resulting in NF-κB activation, COX-2 mRNA induction, COX-2 protein production and PGE_2 synthesis. This chain of events might contribute to the pathogenesis of diabetic nephropathy.

高血糖增加牛内皮细胞线粒体电子传递链中的活性氧种(ROS)的产生(Nishikawa等人。2000；自然404：787-90)。由于一些研究假设前列腺素(PGs)在早期糖尿病的肾小球高滤过中起作用，我们评估了线粒体ROS对培养的人肾小球系膜细胞(HMC)诱导的环氧合酶-2(COX-2)表达的影响。我们首次证实，与5.6 mM葡萄糖相比，30 mM葡萄糖孵育的HMC显著增加了C0X-2mRNA，而不是COX-1mRNA。同样，HMC与30 mM葡萄糖孵育后，线粒体膜电位、细胞内ROS生成、COX-2蛋白表达和PGE_2合成显著增加，这些作用可被线粒体代谢抑制剂TTFA或CCCP完全抑制，或被解偶联蛋白-1或锰超氧化物歧化酶过度表达所抑制。体内研究证实，糖尿病小鼠肾小球COX-2mRNA和蛋白表达增加。此外，高血糖诱导COX-2基因启动子的激活，而该启动子区域的两个核因子κB(核因子κB)结合位点的突变完全取消了该启动子。我们的结果表明，高血糖增加了线粒体ROS的产生，导致了NF-κB的激活，COX-2mRNA的诱导，COX-2蛋白的产生和PGE2的合成。这一系列事件可能参与了糖尿病肾病的发病机制。

项目成果

荒木栄一, 西川武志: "細胞内酸化ストレスと糖尿病性合併症"プラクティス. 20. 9-11 (2003)

Eiichi Araki、Takeshi Nishikawa：“细胞内氧化应激和糖尿病并发症”实践。20. 9-11 (2003)。

Kiritoshi S, Nishikawa T, Sonoda K et al.: "Reactive oxygen species from mitochondria induce cyclooxygenase-2 gene expression in human mesangial cells : potential role in diabetic nephropathy."Diabetes. 52. 2570-2577 (2003)

Kiritoshi S、Nishikawa T、Sonoda K 等人：“线粒体中的活性氧诱导人类系膜细胞中的环氧合酶 2 基因表达：在糖尿病肾病中的潜在作用。”糖尿病。

Sakai K, Matsumoto K, Nishikawa T et al.: "Mitochondrial reactive oxygen species reduce insulin secretion by pancreatic β-cells."Biochem Biophys Res Commun. 300. 216-222 (2003)

Sakai K、Matsumoto K、Nishikawa T 等人：“线粒体活性氧减少胰腺 β 细胞的胰岛素分泌。”Biochem Biophys Res Commun. 300. 216-222 (2003)

西川武志, 園田和洋, 久木留大介, 荒木栄一: "酸化ストレスと糖尿病合併症"現代医療. 35. 65-71 (2003)

Takeshi Nishikawa、Kazuhiro Sonoda、Daisuke Kukidome、Eiichi Araki：“氧化应激和糖尿病并发症”现代医学。 35. 65-71 (2003)

Kiritoshi S, Nishikawa T, Sonoda K et al.: "Reactive oxygen species from mitochondria induce cyclooxygenase-2 gene expression in human mesangial cells : potential role in diabetic nephropathy"Diabetes. 52. 2570-2577 (2003)

Kiritoshi S、Nishikawa T、Sonoda K 等人：“线粒体中的活性氧诱导人系膜细胞中的环氧合酶 2 基因表达：在糖尿病肾病中的潜在作用”糖尿病。