Matsunaga K, Furchgott R F
Department of Pharmacology, State University of New York, Brooklyn.
J Pharmacol Exp Ther. 1991 Dec;259(3):1140-6.
In a perfusion-superfusion system, an endothelium-free ring of rabbit thoracic aorta precontracted with phenylephrine was used to bioassay relaxing activity. As previously reported, when NaNO2 (1 mM) was added to the standard Krebs' solution (37 degrees C oxygenated, containing 30 microM EDTA), irradiation of the solution with long wavelength ultraviolet light, while it was perfusing through a glass tube proximal to the bioassay ring, resulted in a small degree of relaxation of the ring, and this relaxation was markedly potentiated when superoxide dismutase (SOD, 10 U/ml) was also present in the solution. The full potentiation by SOD of the relaxation produced by photoactivation of NO2- was matched by cytochrome c (30 microM), MnCl2 (30 microM) and CuCl2 (100 microM), all of which are scavengers of superoxide (O2-). No potentiation was produced by catalase (1,000 U/ml), ZnCl2 (100 microM) and boiled SOD. Also, CuCl2, when its concentration was no greater than that of the EDTA in the Krebs' solution, failed to potentiate the relaxation. These results are consistent with our earlier conclusion that the relaxant produced by irradiation of NO2- is NO and that it is normally inactivated rapidly by O2- also present in the irradiated perfusion fluid. When EDTA was omitted from the perfusing Krebs' solution, photoactivation of NO2- in the solution produced a very large relaxation of the bioassay ring, which was not enhanced by addition of SOD or other O2- scavengers.(ABSTRACT TRUNCATED AT 250 WORDS)
在一个灌注-再灌注系统中,使用预先用去氧肾上腺素预收缩的兔胸主动脉无内皮环来生物测定舒张活性。如先前报道,当向标准Krebs溶液(37℃充氧,含30μM EDTA)中加入NaNO₂(1 mM)时,在溶液流经生物测定环近端的玻璃管时用长波长紫外线照射该溶液,会导致环出现轻微舒张,并且当溶液中同时存在超氧化物歧化酶(SOD,10 U/ml)时,这种舒张会显著增强。细胞色素c(30μM)、MnCl₂(30μM)和CuCl₂(100μM)(均为超氧阴离子(O₂⁻)清除剂)对NO₂⁻光活化产生的舒张作用的增强程度与SOD完全相同。过氧化氢酶(1000 U/ml)、ZnCl₂(100μM)和煮沸的SOD则不会产生增强作用。此外,当CuCl₂的浓度不高于Krebs溶液中EDTA的浓度时,也无法增强舒张作用。这些结果与我们早期的结论一致,即NO₂⁻照射产生的舒张剂是NO,且它通常会被照射的灌注液中也存在的O₂⁻迅速灭活。当灌注的Krebs溶液中省略EDTA时,溶液中NO₂⁻的光活化会使生物测定环产生非常大的舒张,添加SOD或其他O₂⁻清除剂并不会增强这种舒张。(摘要截短于250字)