An approach to understand the interaction of lipoproteins and vascular endothelial cells

了解脂蛋白和血管内皮细胞相互作用的方法

• 批准号: 04454266
• 负责人: YOKOYAMA Mitsuhiro
• 金额: $ 4.29万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454266/
• 关键词: vascular endothelial cells; endothelium-derived relaxing factor; low density lipoprotein; high density lipoprotein; nitric synthase; protein kinase C; lysophosphatidylcholine; intracellular calcium concentration

(1) Effect of lipoproteins and lysolipids on endothelium-dependent relaxationWe have demonstrated that lysophosphatidylcholine (LPC) in oxidized low density lipoprotein (ox-LDL) inhibits the synthesis of EDRF/NO in endothelial cells. LPC and ox-LDL inhibit the intracellular signal transduction which links the receptor stimulation to biosynthesis of the second massenger in endothelial cells, leading to an endothelial defect in the generation at appropriate intracellular calcium signals nesessary for activation of NO synthase. HDL reverses the ox-LDL-induced impairment of endothelium-dependent relaxation by removing LPC from ox-LDL(2) Regulation of endothelial NO synthase (NOS)We have purified endothelial NOS from cultured bovine aortic endothelial cells (BAECs). Endothelial NOS was mainly regulated Ca^<2+>/calmodulin in the presence of NADPH,FAD and BH_4 as cofactors. NOS activity was enhanced approximately up to three fold by PC, LPC and phosphatidylethanolamice. NOS mRNA expression was investigated in BAECs. LPC or ox-LDL enhanced NOS mRNA expression.Interferon alpha/beta increased NOS mRNA expression, but TNF-alpha inhibited it.(3) Regulation of inducible NOS (iNOS) mRNA expression in vascular smooth muscle cells. interferon gamma, TNF alpha and interleukin-1beta each markedly increased mRNA and protein levels of iNOS in parallel with the production of nitrite. TGF-beta significantly inhibited the increase in protein levels caused by these cytokines.

(1)脂蛋白和溶脂对内皮依赖性松弛的影响我们证明氧化型低密度脂蛋白(OX-LDL)中的溶血磷脂酰胆碱(LPC)抑制内皮细胞EDRF/NO的合成。LPC和OX-LDL抑制细胞内信号转导，该信号转导将受体刺激与内皮细胞第二粘合剂的生物合成联系起来，导致内皮细胞在激活NO合成酶所需的适当细胞内钙信号的产生上出现缺陷。高密度脂蛋白通过去除氧化低密度脂蛋白中的LPC而逆转氧化型低密度脂蛋白所致的内皮依赖性松弛损伤(2)内皮型一氧化氮合酶(NOS)的调节我们从培养的牛主动脉内皮细胞(BAECs)中纯化了内皮型一氧化氮合酶。在NADPH、FAD和BH_4的协同作用下，内皮细胞的NOS主要受Ca~(2+)和Gt~(2+)/CaM的调节。PC、LPC和磷脂酰乙醇胺可使一氧化氮合酶活性增加近3倍。观察BAECs中一氧化氮合酶基因的表达。低密度脂蛋白或氧化低密度脂蛋白促进一氧化氮合酶的表达，干扰素α/β可促进一氧化氮合酶的表达，而肿瘤坏死因子α可抑制其表达。(3)血管平滑肌细胞诱导型一氧化氮合酶(INOS)表达的调节。干扰素、肿瘤坏死因子α和白介素1β均显著增加iNOS的mRNA和蛋白水平，同时亚硝酸盐的产生也随之增加。转化生长因子-β显著抑制这些细胞因子引起的蛋白质水平的增加。

项目成果

K.Hirata: "Impaired vasodilatory response to atrial natriuretic peptide during atherosclerosis progression." Arteriosclerosis and Thrombosis. 12. 99-105 (1992)

K.Hirata：“在动脉粥样硬化进展过程中，心房钠尿肽的血管舒张反应受损。”

Y.Matsuda: "High density lipoprotein reveses inhibitory effect of oxidized low density lipoprotein on endothelium-dependent arterial relaxation." Circ Res. 72. 1103-1109 (1993)

Y.Matsuda：“高密度脂蛋白可逆转氧化低密度脂蛋白对内皮依赖性动脉舒张的抑制作用。”

N.Inoue: "Lysophosphatidylcholine inhibits badykinin-induced phosphoinositide hydrolysis and calcium transients in cultured bovine aortic endothelial cells" Circ Res. 71. 1410-1421 (1992)

N.Inoue：“溶血磷脂酰胆碱抑制培养的牛主动脉内皮细胞中坏激肽诱导的磷酸肌醇水解和钙瞬变”Circ Res。

M.Koide: "Expression of nitric oxide synthase by cytokines in vascular smooth muscle cells" Hypertension. 23. 45-48 (1994)

M.Koide：“血管平滑肌细胞中细胞因子表达一氧化氮合酶”高血压。

N.Inoue: "Lysophosphatidylcholine inhibits bradykinin-induced phosphoinositide hydrolysis and calcium transients in cultured bovine aortic endothelial cells." Circ Res. 71. 1410-1421 (1992)

N.Inoue：“溶血磷脂酰胆碱可抑制培养的牛主动脉内皮细胞中缓激肽诱导的磷酸肌醇水解和钙瞬变。”