Joo Nam Soo, Saenz Yamil, Krouse Mauri E, Wine Jeffrey J
Cystic Fibrosis Research Laboratory, Stanford University, Stanford, California 94305-2130, USA.
J Biol Chem. 2002 Aug 2;277(31):28167-75. doi: 10.1074/jbc.M202712200. Epub 2002 May 13.
Secretion rates of >700 individual glands in isolated tracheal mucosa from 56 adult pigs were monitored optically. "Basal" secretion of 0.7 +/- 0.1 nl x min(-1) gland(-1) was observed 1-9 h post-harvest but was near zero on day 2. Secretion to carbachol (10 microm) peaked at 2-3 min and then declined to a sustained phase. Peak secretion was 12.4 +/- 1.1 nl x min(-1) gland(-1); sustained secretion was approximately one-third of peak secretion. Thapsigargin (1 microm) increased secretion from 0.1 +/- 0.05 to 0.7 +/- 0.2 nl x min(-1) gland(-1); thapsigargin did not cause contraction of the trachealis muscles. Isoproterenol and phenylephrine (10 microm each) were ineffective, but vasoactive intestinal peptide (1 microm) and forskolin (10 microm) each produced sustained secretion of 1.0 +/- 0.5 and 1.7 +/- 0.2 nl x min(-1) gland(-1), respectively. The density of actively secreting glands was 1.3/mm(2). Secretion to either carbachol or forskolin was inhibited (approximately 50%) by either bumetanide or HCO(3)(-) removal and inhibited approximately 90% by the combined treatments. Mucus secreted in response to carbachol or forskolin was acidic by approximately 0.2 pH units relative to the bath and remained acidic by approximately 0.1 pH units after bumetanide. The strong secretory response to vasoactive intestinal peptide, the acidity of cAMP-stimulated mucus, and its inhibition by bumetanide were unexpected.
对56只成年猪分离出的气管黏膜中700多个腺体的分泌速率进行了光学监测。收获后1 - 9小时观察到“基础”分泌量为0.7±0.1 nl·min⁻¹·腺⁻¹,但在第2天接近零。对卡巴胆碱(10 μmol)的分泌在2 - 3分钟达到峰值,然后下降到持续阶段。峰值分泌量为12.4±1.1 nl·min⁻¹·腺⁻¹;持续分泌量约为峰值分泌量的三分之一。毒胡萝卜素(1 μmol)使分泌量从0.1±0.05增加到0.7±0.2 nl·min⁻¹·腺⁻¹;毒胡萝卜素未引起气管肌收缩。异丙肾上腺素和去氧肾上腺素(各10 μmol)无效,但血管活性肠肽(1 μmol)和福斯高林(10 μmol)分别产生了1.0±0.5和1.7±0.2 nl·min⁻¹·腺⁻¹的持续分泌。活跃分泌腺体的密度为1.3/mm²。布美他尼或去除HCO₃⁻可使对卡巴胆碱或福斯高林的分泌抑制约50%,联合处理则抑制约90%。对卡巴胆碱或福斯高林反应分泌的黏液相对于浴液酸性约为0.2个pH单位,布美他尼处理后仍酸性约0.1个pH单位。对血管活性肠肽的强烈分泌反应、[cAMP]i刺激的黏液的酸性及其被布美他尼抑制是出乎意料的。