Endothelial cell ER stress and Ca^<2+> signaling pathway

内皮细胞ER应激与Ca^2信号通路

• 批准号: 14570652
• 负责人: WATANABE Hirosi
• 金额: $ 2.18万
• 依托单位: Hamamatsu University School of Medicine
• 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570652/
• 关键词: apoptosis; endothelial cell; calcium; ER stress; caspase; Akt; PI3キナーゼ

Apoptosis of endothelial cells (ECs) is now regarded to be an initial step inducing atherosclerosis. Recent studies have reported that the depletion of endoplasmic reticulum (ER) Ca^<2+> stores plays an important role in apoptosis. Caspase-12 is a key signal to lead ER stress-induced apoptosis. However, it is not known whether the depletion of ER Ca^<2+> is linked to caspase-12 signaling in ECs. Here we have investigated the interaction of Ca^<2+> signaling and caspase-12 cleavage in apoptosis of cultured porcine aortic ECs. Cytosolic Ca^<2+> concentration ([Ca^<2+>]i) was measured using fura-2/AM. Apoptosis was assessed by DNA ladder formation, and cleavage of caspase-12 by Western blotting. Thapsigargin (6μM), an inhibitor of the ER-associated Ca^<2+>-ATPase, increased [Ca^<2+>]i (F340/F380 ratio 0.717±0.137 to 6.133±0.710 : p<0.001) and induced persistent ER calcium depletion, cleavage of caspase-12 and apoptosis. Bradykinin (10 nM) increased [Ca^<2+>]i (F340/F380 ratio 0.717±0.053 to 6.133±0.528 : p<0.001) but induced neither cleavage of caspase-12 nor apoptosis. However, when ECs were treated with BAPTA/AM (100μM), BK caused the Ca^<2+> depletion of ER and apoptosis without the cleavage of caspase-12. Moreover non-selective caspase inhibitor, zVAD-fmk (100μM), inhibited apoptosis and cleavage of caspase-12 in TG-stimulated ECs, and calpain inhibitor, MDL 28170 (120μM), inhibited cleavage of caspase-12 but not inhibited apoptosis. These results suggested that the increase of [Ca^<2+>]i did not play an important role in inducing apoptosis in ECs, and ER Ca^<2+> depletion induced apoptosis, which is independent from caspase-12 linked signaling pathway.

内皮细胞凋亡被认为是动脉粥样硬化形成的第一步。最近的研究表明，内质网（ER）钙库的耗竭在细胞凋亡中起重要作用。Caspase-12是导致内质网应激诱导凋亡的关键信号。然而，目前还不清楚ER Ca^<2+>的耗竭是否与EC中的caspase-12信号转导有关。在此，我们研究了Ca^2+信号转导和caspase-12切割在培养的猪主动脉内皮细胞凋亡中的相互作用。用Fura-2/AM测定胞浆Ca^2+浓度（[Ca^2+]i）。DNA梯状条带形成法检测细胞凋亡，Western印迹法检测caspase-12的裂解。Thapsigargin（6μM）是一种ER相关Ca^<2+>-ATP酶的抑制剂，可增加[Ca^<2+>]i（F340/F380比值为0.717±0.137至6.133±0.710：p<0.001），并诱导持续的ER钙耗竭、caspase-12裂解和细胞凋亡。缓激肽（10 nM）增加[Ca^<2+>]i（F340/F380比值为0.717±0.053至6.133±0.528：p<0.001），但既不诱导caspase-12裂解，也不诱导凋亡。然而，当用BAPTA/AM（100μM）处理EC时，BK引起ER的Ca^<2+>耗竭和凋亡，而不引起caspase-12的裂解。此外，非选择性caspase抑制剂zVAD-favor（100μM）可抑制TG刺激的EC凋亡和caspase-12的裂解，而钙蛋白酶抑制剂MDL 28170（120μM）可抑制caspase-12的裂解，但不抑制凋亡。上述结果提示，[Ca^<2+>]i升高在诱导内皮细胞凋亡中不起重要作用，ER Ca^<2+>耗竭诱导内皮细胞凋亡不依赖于caspase-12相关信号通路。

项目成果

Effects of cytochrome P450 inhibitors on agonist-induced Ca2+ responses and production of NO and PGI2 in vascular endothelial cells

细胞色素 P450 抑制剂对激动剂诱导的 Ca2+ 反应以及血管内皮细胞中 NO 和 PGI2 产生的影响

• DOI: 10.1023/a:1024136318779
• 发表时间: 2003
• 期刊: Molecular and Cellular Biochemistry
• 影响因子: 4.3
• 作者: K. Takeuchi;Hiroshi Watanabe;Q. Tran;Mariko Ozeki;A. Uehara;H. Katoh;H. Satoh;H. Terada;K. Ohashi;H. Hayashi
• 通讯作者: H. Hayashi

Nitric oxide : inhibitory effects on endothelial cell calcium signaling, prostaglandin 12 production and nitric oxide synthase expression.

一氧化氮：对内皮细胞钙信号传导、前列腺素 12 产生和一氧化氮合酶表达的抑制作用。

• 发表时间: 2004
• 期刊: CARDIOVASCULAR RESEARCH 62
• 作者: Takeuchi K;Watanabe H;Tran QK;Ozeki M;Sumi D;Hayashi T;Iguchi A;Ignarro LJ;Ohashi K;Hayashi H
• 通讯作者: Hayashi H

Akt and Ca2+ signaling in endothelial cells

内皮细胞中的 Akt 和 Ca2 信号传导

• DOI: 10.1023/b:mcbi.0000021369.17958.f4
• 发表时间: 2004
• 期刊: Molecular and Cellular Biochemistry
• 影响因子: 4.3
• 作者: Mariko Ozeki;Hiroshi Watanabe;Jinghui Luo;T. Nakano;K. Takeuchi;Y. Kureishi;Masa;T. Nakano;K. Ohashi;H. Hayashi
• 通讯作者: H. Hayashi

Nitric oxide:: inhibitory effects on endothelial cell calcium signaling, prostaglandin I2 production and nitric oxide synthase expression

• DOI: 10.1016/j.cardiores.2003.12.028
• 发表时间: 2004-04-01
• 期刊: CARDIOVASCULAR RESEARCH
• 影响因子: 10.8
• 作者: Takeuchi, K;Watanabe, H;Hayashi, H
• 通讯作者: Hayashi, H

Modification of sarcoplasmic reticulum (SR) Ca^<2+> release by FK506 induces defective excitationcontraction coupling only when SR Ca^<2+> recycling is disturbed

仅当 SR Ca^<2> 再循环受到干扰时，FK506 对肌浆网 (SR) Ca^<2> 释放的修饰才会诱导缺陷的兴奋收缩耦合

• 发表时间: 2005
• 期刊: Can.J.Physiol.Pharmacol. 83
• 作者: Yoshihara S;Satoh H;Saotome M;Katoh H;Terada H;Watanabe H;Hayashi H
• 通讯作者: Hayashi H