使用克拉克型电极对牛灌注眼小梁网中的一氧化氮进行实时直接测量。
Real-time direct measurement of nitric oxide in bovine perfused eye trabecular meshwork using a clark-type electrode.
作者信息
Millar J C
机构信息
Department of Biology & Health Science, Faculty of Science, Technology & Design, University of Luton, Park Square Campus, Luton, England.
出版信息
J Ocul Pharmacol Ther. 2003 Aug;19(4):299-313. doi: 10.1089/108076803322279363.
NO was detected in bovine trabecular meshwork (TM). Bovine eyes were perfused (posterior ciliary artery). In some eyes (operated eyes) a NO electrode was inserted adjacent to the TM (scleral flap). Vascular perfusion/intraocular pressures (VPP/IOP) were recorded. In operated eyes, epinephrine (1 nM-100 microM) increased NO (maximally 979.9 +/- 117.6 nM, mean +/- SDM). Timolol (1 mM) shifted the epinephrine-NO concentration-response curve rightward (2.94 log units) without significantly changing the maximal response (903.0 +/- 67.7 nM, mean +/- SDM). The non-selective NO synthase (NOS) inhibitor L-NMMA (100 microM) virtually abolished the NO response to epinephrine. L-NMMA alone (1 microM-100 microM) significantly reduced tonic NO generation (maximally 109.5 +/- 24.9 nM, mean +/- SDM), whereas timolol alone (1 microM-1 mM) had no effect. In unoperated eyes, epinephrine (1 nM-100 microM) reduced IOP (maximally 2.56 +/- 0.64 mmHg, mean +/- SDM). Epinephrine (100 microM) mildly increased VPP (4.6 +/- 1.3 mmHg, mean +/- SDM). Baseline aqueous humor formation rate (11.5 +/- 3.2 microl/min, mean +/- SDM) was unaffected. Effluent perfusate (effusate) total NO(2)(-) was determined by enzymatically reducing all NO(3)(-) to NO(2)(-), then assessing resultant NO(2)(-) (Griess assay). Epinephrine (1 nM-1 microM) increased effusate NO(2)(-) (maximally 15.8 +/- 4.9 microM, mean +/- SDM). Timolol (1 mM) reduced, and L-NMMA (100 microM) virtually abolished effusate NO(2)(-) response to epinephrine. L-NMMA alone (1 microM-100 microM) reduced tonic effusate NO(2)(-) (maximally from 5.8 +/- 1.6 microM to 1.1 +/- 0.9 microM, mean +/- SDM), whereas timolol alone (1 microM-1 mM) had no effect. NO is generated tonically in bovine TM and increases in response to epinephrine.
在牛小梁网（TM）中检测到一氧化氮（NO）。对牛眼进行灌注（通过睫状后动脉）。在一些眼睛（手术眼）中，将NO电极插入靠近TM（巩膜瓣）的位置。记录血管灌注/眼内压（VPP/IOP）。在手术眼中，肾上腺素（1 nM - 100 microM）可增加NO（最大增加至979.9 +/- 117.6 nM，平均值 +/- 标准差）。噻吗洛尔（1 mM）使肾上腺素 - NO浓度 - 反应曲线向右移动（2.94个对数单位），而最大反应无显著变化（903.0 +/- 67.7 nM，平均值 +/- 标准差）。非选择性一氧化氮合酶（NOS）抑制剂L - NMMA（100 microM）几乎完全消除了对肾上腺素的NO反应。单独使用L - NMMA（1 microM - 100 microM）可显著降低持续性NO生成（最大降低至109.5 +/- 24.9 nM，平均值 +/- 标准差），而单独使用噻吗洛尔（1 microM - 1 mM）则无作用。在未手术的眼睛中，肾上腺素（1 nM - 100 microM）可降低眼压（最大降低2.56 +/- 0.64 mmHg，平均值 +/- 标准差）。肾上腺素（100 microM）可轻度增加VPP（4.6 +/- 1.3 mmHg，平均值 +/- 标准差）。基线房水生成率（11.5 +/- 3.2微升/分钟，平均值 +/- 标准差）未受影响。通过酶促将所有硝酸根（NO3(-)）还原为亚硝酸根（NO2(-)），然后评估生成的NO2(-)（Griess法）来测定流出灌注液（流出液）中的总NO2(-)。肾上腺素（1 nM - 1 microM）可增加流出液中的NO2(-)（最大增加至15.8 +/- 4.9 microM，平均值 +/- 标准差）。噻吗洛尔（1 mM）可降低，而L - NMMA（100 microM）几乎完全消除了流出液对肾上腺素的NO2(-)反应。单独使用L - NMMA（1 microM - 100 microM）可降低持续性流出液中的NO2(-)（最大从5.8 +/- 1.6 microM降至1.1 +/- 0.9 microM，平均值 +/- 标准差），而单独使用噻吗洛尔（1 microM - 1 mM）则无作用。在牛TM中持续性生成NO，并对肾上腺素产生反应而增加。
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