Tosco M, Orsenigo M N, Gastaldi G, Faelli A
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, via Celoria 26, I-20133 Milan, Italy.
Biochim Biophys Acta. 1998 Jul 17;1372(2):323-30. doi: 10.1016/s0005-2736(98)00068-6.
During bicarbonate absorption in rat jejunum, a Cl/HCO3 exchanger mediates bicarbonate extrusion across the basolateral membrane of the enterocyte. Previous studies demonstrated that anion antiport exhibits a particular behaviour: its activity is positively affected by the presence of sodium, but the cation is not translocated by the carrier protein. In view of the particular features of the jejunal Cl/HCO3 antiporter, first we performed a pharmacological characterisation of the transport protein using various Cl channels blockers. Then, since it is well known that anion exchangers play a substantial role in cell pH regulation, we investigated the possible involvement of jejunal basolateral Cl/HCO3 antiporter in intracellular pH maintenance. The sensitivity of the exchanger to pH was investigated by measuring 36Cl uptake into basolateral membrane vesicles either varying simultaneously intra- and extravesicular pH, or presetting at 7.4 external pH and varying only the internal one. Experiments were performed both in the absence and in the presence of Na. In all the tested conditions, uptake peaked at pH of about 7. 3-7.4 and then decreased, suggesting that the main function of Cl/HCO3 exchanger is related to HCO3 absorption rather than to intracellular pH control. Since pH-regulating mechanisms counteracting acidification are well known in the jejunal enterocyte, we investigated how it regulates pH after alkalinisation of the cytosol. We tested both basolateral and brush border membrane vesicles for the presence of a K/H exchanger, but we could not give evidence for its presence by means of 86Rb uptake experiments. In conclusion, the jejunal enterocyte seems to lack a mechanism counteracting cellular alkalinisation: the main purpose of pH homeostasis might be to hinder acidification of the cytosol due to influx of protons and production of acid by the metabolism.
在大鼠空肠碳酸氢盐吸收过程中,一种Cl/HCO3交换体介导碳酸氢盐穿过肠上皮细胞的基底外侧膜排出。先前的研究表明,阴离子反向转运表现出一种特殊行为:其活性受到钠离子存在的正向影响,但阳离子不会被载体蛋白转运。鉴于空肠Cl/HCO3反向转运体的特殊特征,我们首先使用各种Cl通道阻滞剂对转运蛋白进行了药理学特性分析。然后,由于众所周知阴离子交换体在细胞pH调节中起重要作用,我们研究了空肠基底外侧Cl/HCO3反向转运体在维持细胞内pH方面的可能作用。通过测量36Cl进入基底外侧膜囊泡的摄取量来研究交换体对pH的敏感性,摄取量的测量通过同时改变囊泡内外的pH值,或预设外部pH值为7.4并仅改变内部pH值来进行。实验在有无钠离子的情况下均进行。在所有测试条件下,摄取量在pH约为7.3 - 7.4时达到峰值,然后下降,这表明Cl/HCO3交换体的主要功能与HCO3吸收有关,而非细胞内pH控制。由于在空肠肠上皮细胞中,对抗酸化的pH调节机制是众所周知的,我们研究了细胞溶质碱化后它如何调节pH。我们测试了基底外侧膜囊泡和刷状缘膜囊泡中是否存在K/H交换体,但通过86Rb摄取实验无法证明其存在。总之,空肠肠上皮细胞似乎缺乏对抗细胞碱化的机制:pH稳态的主要目的可能是阻止由于质子内流和代谢产生酸而导致的细胞溶质酸化。