Yu J, Ong B Y, Zheng J, Cragoe E J, Bose D, Bose R
Department of Pharmacology, University of Manitoba, Winnipeg, Canada.
Am J Physiol. 1996 Nov;271(5 Pt 2):H1912-7. doi: 10.1152/ajpheart.1996.271.5.H1912.
We simultaneously measured intracellular pH (pHi) and isometric tension in canine basilar arteries. pHi was calculated from the ratio of fluorescence intensities at 540 nm of exciting wavelengths of 500 and 440 nm in the presence of 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). pHi was 7.33 +/- 0.02 in Krebs-Henseleit solution (pH of 7.4 at 37 degrees C). Application of the anion exchange blocker 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) decreased the resting pHi by 0.25 pH units. Increasing extracellular pH (pHo) [by decreasing CO2 tension (PCO2)] from 7.4 to 7.8 increased pHi by 0.38 pH units and increased tension by 2.28 +/- 0.21 mN. Decreasing pHo from 7.8 to 7.4 (by increasing PCO2) restored the pHi and muscle tension to their baseline levels. SITS inhibited the increase in pHi and isometric tension in response to the increase in pHo in an endothelium-independent fashion. The Na+/H+ exchange blockers, amiloride or 5-(N-methyl-N-guanidinocarbonylmethyl)-amiloride, did not affect the pHo or tension changes. The results suggest that in the range of pH tested, anion exchange is more important than Na+/H+ exchange in the regulation of pHi and mechanical tone in the basilar artery.
我们同时测量了犬基底动脉的细胞内pH值(pHi)和等长张力。在2',7'-双(羧乙基)-5(6)-羧基荧光素(BCECF)存在的情况下,根据激发波长为500和440 nm时540 nm处荧光强度的比值计算pHi。在Krebs-Henseleit溶液(37℃时pH为7.4)中,pHi为7.33±0.02。应用阴离子交换阻滞剂4-乙酰胺基-4'-异硫氰基芪-2,2'-二磺酸(SITS)可使静息pHi降低0.25个pH单位。将细胞外pH值(pHo)[通过降低二氧化碳张力(PCO2)]从7.4提高到7.8,可使pHi升高0.38个pH单位,并使张力升高2.28±0.21 mN。将pHo从7.8降低到7.4(通过提高PCO2)可使pHi和肌肉张力恢复到基线水平。SITS以不依赖内皮的方式抑制了因pHo升高而引起的pHi和等长张力的增加。钠/氢交换阻滞剂氨氯地平或5-(N-甲基-N-胍基羰基甲基)-氨氯地平不影响pHo或张力变化。结果表明,在所测试的pH范围内,阴离子交换在基底动脉pHi和机械张力的调节中比钠/氢交换更重要。