Tobey N A, Koves G, Orlando R C
Department of Medicine, Tulane University School of Medicine, and the Veterans Administration Medical Center, New Orleans, Louisiana, USA.
Am J Gastroenterol. 1998 Nov;93(11):2075-81. doi: 10.1111/j.1572-0241.1998.00596.x.
The human esophagus is regularly exposed to refluxed gastric acid. Therefore, its epithelial cells require for survival a means of extruding excess H+ from the cytoplasm. Because Na+/H+ exchange activity has been observed in many mammalian cell types, including that of rabbit esophagus, we sought its presence in human esophageal epithelium.
Human esophageal epithelial cells derived from endoscopic biopsy specimens or surgical esophagectomy specimens were grown in primary culture and loaded with the fluorescent dye, 2'7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, to monitor intracellular pH (pHi).
Resting pHi in bicarbonate-free N'-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid was 7.5 +/- 0.03 (n = 50). Acidification using the NH4Cl prepulse technique lowered pHi by 0.6 +/- 0.02 pH units, with recovery ensuing at an initial rate of 0.09 +/- 0.04 pH units/min. Notably, the rate of recovery was faster the more acidic the pHi, and recovery was abolished by amiloride or replacement with an Na+-free buffer. Acidification by lowering pHo with HCl resulted in a similarly rapid rate of return as with the NH4Cl technique, and resting cells acidified by 0.17 +/- 0.02 pH units/5 min upon exposure to amiloride.
Human esophageal cells possess an H+-extruding mechanism consistent with an Na+/H+ exchanger. This mechanism is active in resting cells, adapts to the degree of pHi lowering, and extrudes H+ efficiently whether loaded by intracellular or extracellular means, making it well suited for epithelial defense against acid injury.
人类食管经常暴露于反流的胃酸中。因此,其上皮细胞要存活就需要一种将多余的H⁺从细胞质中排出的方式。由于在包括兔食管细胞在内的许多哺乳动物细胞类型中都观察到了Na⁺/H⁺交换活性,我们探寻其在人类食管上皮中的存在情况。
从内镜活检标本或手术切除的食管标本中获取的人类食管上皮细胞进行原代培养,并加载荧光染料2'7'-双(羧乙基)-5(6)-羧基荧光素以监测细胞内pH值(pHi)。
在不含碳酸氢盐的N'-2-羟乙基哌嗪-N'-2-乙磺酸中,静息pHi为7.5±0.03(n = 50)。使用氯化铵预脉冲技术酸化可使pHi降低0.6±0.02个pH单位,随后以0.09±0.04个pH单位/分钟的初始速率恢复。值得注意的是,pHi酸性越强,恢复速率越快,而氨氯吡咪或用无钠缓冲液替代可消除恢复过程。用盐酸降低细胞外pH值进行酸化导致的恢复速率与氯化铵技术相似,暴露于氨氯吡咪后,静息细胞每5分钟酸化0.17±0.02个pH单位。
人类食管细胞拥有一种与Na⁺/H⁺交换器一致的H⁺排出机制。该机制在静息细胞中具有活性,能适应pHi降低的程度,并且无论是通过细胞内还是细胞外方式加载H⁺,都能有效地将其排出,使其非常适合上皮细胞抵御酸损伤。