The effects of hypoxia on pHi in porcine coronary artery endothelium and smooth muscle. A novel method for measurements in endothelial cells in situ.
The effects of hypoxia on pHi in porcine coronary artery endothelium and smooth muscle. A novel method for measurements in endothelial cells in situ.
复制标题
缺氧对猪冠状动脉内皮和平滑肌pHi的影响。
DOI：
10.1161/01.res.80.1.21
复制
发表时间：
1997
影响因子：
20.1
通讯作者：
Paul,RJ
中科院分区：
文献类型：
--
作者：
Foy,RA;Shimizu,S;Paul,RJ
缺氧条件下猪冠状动脉内皮依赖性舒张减弱。最近的证据还表明，pH值可能调节内皮源性舒张因子的释放。我们检验了缺氧诱导的内皮依赖性舒张衰减是由pHi改变介导的这一假设。我们开发了一种新的加载表面细胞的方法，从而可以在原位测量完整的猪冠状动脉内皮细胞的pH值。用荧光指示剂BCECF-AM选择性负载动脉环段的内皮细胞。通过共聚焦显微镜验证内皮细胞层的差异负载。用Photon Technology International荧光分光光度计测量原位内皮细胞和内皮剥脱动脉的pH。内皮功能的完整性进行了评估的内皮依赖性舒张物质在一对相邻的环。在pH测量的实验方案中，用生理碳酸氢盐缓冲液（pH 7.4）灌注制剂，用KCl（29 mmol/L）刺激，然后进行缺氧和复氧。6条完整动脉内皮细胞的平均基础pH值为6.92±0.07。向灌注培养基中添加KCl使pH降低（P= 0.025）至6.79±0.07。随后用N2鼓泡使pHi增加至7.00±0.06（P= 0.009），这被再给氧逆转。与原位内皮细胞相比，KCl或缺氧均未显著改变平滑肌的pH值（7.24±0.06，n=5）。这种差异行为证实了共聚焦数据表明差异染料加载。因此，这些数据表明，氧敏感性改变的pH值可能是一个重要的机制，在内皮细胞的信号转导。
Endothelium-dependent relaxation of porcine coronary arteries is attenuated under hypoxic conditions. Recent evidence also indicates that pHimay modulate the release of the endothelium-derived relaxing factor. We tested the hypothesis that hypoxia-induced attenuation of endothelium-dependent relaxation is mediated by alterations in pHi. We developed a novel method for loading surface cells, whereby endothelial cell pHicould be measured in situ on the intact porcine coronary artery. Endothelial cells of arterial ring segments were selectively loaded with the fluorescent indicator BCECF-AM. Differential loading of the endothelial cell layer was verified by confocal microscopy. pHiof the endothelial cells in situ and of endothelium-denuded arteries was measured with a Photon Technology International spectrofluorimeter. The functional integrity of the endothelium was assessed by the endothelium-dependent relaxation to substance P in a paired adjacent ring. In the experimental protocol for pHimeasurements, preparations were perfused with a physiological bicarbonate buffer (pH 7.4), stimulated with KCl (29 mmol/L), and then subjected to hypoxia and reoxygenation. The mean basal pHiin endothelial cells on the intact six arteries was 6.92±0.07. Addition of KCl to the perfusion medium decreased (P=.025) pHito 6.79±0.07. Subsequent bubbling with N2increased (P=.009) pHito 7.00±0.06, which was reversed by reoxygenation. In contrast to the in situ endothelium, pHiof the smooth muscle was not significantly altered from its basal value of 7.24±0.06 (n=5) by either KCl or hypoxia. This differential behavior corroborated the confocal data indicating differential dye loading. These data thus suggest that oxygen-sensitive alterations in pHimay be an important mechanism of signal transduction in endothelial cells.
登录
查看更多内容
猪冠状动脉内皮介导的舒张对氧和代谢的依赖性。
DOI：
10.1152/ajpheart.1993.265.1.h299
发表时间：
1993
期刊：
The American journal of physiology
影响因子：
--
作者：
Hashimoto,M;Close,LA;Ishida,Y;Paul,RJ
通讯作者：
Paul,RJ
影响因子：
20.1
作者：
HAIGNEY, MCP;MIYATA, H;SILVERMAN, HS
通讯作者：
SILVERMAN, HS
细胞内碱化导致牛主动脉内皮细胞中激动剂敏感库中的 Ca2+ 动员。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Danthuluri,NR;Kim,D;Brock,TA
通讯作者：
Brock,TA
内皮对猪冠状动脉基础张力、激动剂和 O2 敏感性的影响。
DOI：
--
发表时间：
1990
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Ngai,JH;Roth,PS;Paul,RJ
通讯作者：
Paul,RJ
影响因子：
3.3
作者：
MAZZONI, MC;INTAGLIETTA, M;ARFORS, KE
通讯作者：
ARFORS, KE