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首页> 外文期刊>Endothelium: Journal of endothelial cell research >Cigarette smoke and its formaldehyde component inhibit bradykinin-induced calcium increase in pig aortic endothelial cells.
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Cigarette smoke and its formaldehyde component inhibit bradykinin-induced calcium increase in pig aortic endothelial cells.

机译:香烟烟雾及其甲醛成分可抑制缓激肽诱导的猪主动脉内皮细胞钙的增加。

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Bradykinin-induced increase in the intracellular concentration of free calcium evokes an activation of the endothelial nitric oxide synthase (eNOS) enzyme, producing nitric oxide (NO). Cigarette smoke inhibits the eNOS-NO-cGMP signaling pathway. The pathomechanism of this deleterious effect of smoke on NO production is unknown. The aim of this study was to investigate the effect of gas phase smoke trapped in a buffer (smoke buffer, SB) on the bradykinin-induced calcium increase in cultured endothelial cells. FURA-2-AM was used to detect bradykinin-induced calcium increase. A sensitive, fluorescent method using O-phthaldialdehyde was used for the determination of intracellular reduced glutathione (GSH) and protein-thiol levels. SB caused a time- and concentration-dependent inhibition of bradykinin-induced calcium increase. Formaldehyde, a component of SB, inhibited bradykinin-induced calcium increase in concentrations characteristic for SB. SB decreased both the intracellular GSH (0.22 +/- 0.06 vs. 2.23 +/- 0.32 mumol/g protein, SB vs. control, p < .001) and protein-thiol levels (4.98 +/- 0.54 vs. 7.31 +/- 0.97 microEqu GSH/g protein, SB vs. control, p < .05) in the endothelial cells. Intracellular GSH and protein-thiol levels were not changed by 80 microM formaldehyde. GSH (4 mM) prevented the effect of SB (p < .001) and formaldehyde (p < .05) on the bradykinin-induced calcium increase. Our data support the premise that SB inhibits bradykinin-induced calcium increase. This inhibition is partially due to protein-thiol oxidation but may also be caused by the formaldehyde content of SB, which inhibits calcium increase in a protein-thiol-independent manner.
机译:缓激肽诱导的细胞内游离钙浓度增加引起内皮一氧化氮合酶(eNOS)酶的激活,产生一氧化氮(NO)。香烟烟雾抑制eNOS-NO-cGMP信号通路。烟雾对NO产生有害作用的致病机理尚不清楚。这项研究的目的是研究捕获在缓冲液(烟气缓冲液,SB)中的气相烟雾对缓激肽诱导的内皮细胞钙增加的影响。 FURA-2-AM用于检测缓激肽诱导的钙增加。使用邻苯二甲醛的灵敏的荧光方法用于测定细胞内还原型谷胱甘肽(GSH)和蛋白质硫醇的水平。 SB引起缓激肽诱导的钙增加的时间和浓度依赖性抑制。甲醛是SB的一种成分,可抑制缓激肽诱导的SB浓度升高。 SB降低了细胞内GSH(0.22 +/- 0.06 vs. 2.23 +/- 0.32 mumol / g蛋白质,SB相对于对照,p <.001)和蛋白质硫醇水平(4.98 +/- 0.54 vs. 7.31 + / -内皮细胞中的0.97 microEqu GSH / g蛋白,SB对对照,p <.05)。 80 microM甲醛不会改变细胞内GSH和蛋白质硫醇水平。 GSH(4 mM)阻止了SB(p <.001)和甲醛(p <.05)对缓激肽诱导的钙增加的影响。我们的数据支持SB抑制缓激肽诱导的钙增加的前提。这种抑制作用部分是由于蛋白质-硫醇的氧化,但也可能是由于SB的甲醛含量所致,它以蛋白质-硫醇非依赖性的方式抑制钙的增加。

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