Döring F, Walter J, Will J, Föcking M, Boll M, Amasheh S, Clauss W, Daniel H
Institute of Nutritional Sciences, University of Giessen, 35392 Giessen, Germany.
J Clin Invest. 1998 Jun 15;101(12):2761-7. doi: 10.1172/JCI1909.
Delta-aminolevulinic acid (ALA) is the precursor of porphyrin synthesis and has been recently used in vitro and in clinical studies as an endogenous photosensitizer for photodynamic therapy in the treatment of various tumors. For this purpose, ALA is given topically, systemically, or orally. When administered by the oral route, it shows excellent intestinal absorption. ALA is also efficiently reabsorbed in the renal proximal tubule after glomerular filtration. However, the pathways and mechanisms for its transmembrane transport into epithelial cells of intestine and kidney are unknown. Here we demonstrate that ALA uses the intestinal and renal apical peptide transporters for entering into epithelial cells. Kinetics and characteristics of ALA transport were determined in Xenopus laevis ooyctes and Pichia pastoris yeast cells expressing either the cloned intestinal peptide transporter PEPT1 or the renal form PEPT2. By using radiolabeled ALA and electrophysiological techniques in these heterologous expression systems, we established that: (a) PEPT1 and PEPT2 translocate 3H-ALA by saturable and pH-dependent transport mechanisms, (b) that ALA and di-/tripeptides, but not GABA or related amino acids, compete at the same substrate-binding site of the carriers, and (c) that ALA transport is electrogenic in nature as a consequence of H+/ALA cotransport. Reverse transcriptase-PCR analysis performed with specific primers for PEPT1 and PEPT2 in rabbit tissues demonstrates that, in particular, the PEPT2 mRNA is expressed in a variety of other tissues including lung, brain, and mammary gland, which have been shown to accumulate ALA. This suggests that these tissues could take up the porphyrin precusor via expressed peptide transporters, providing the endogenous photosensitizers for efficient photodynamic therapy.
δ-氨基乙酰丙酸(ALA)是卟啉合成的前体,最近已在体外和临床研究中用作内源性光敏剂,用于光动力疗法治疗各种肿瘤。为此,ALA可局部、全身或口服给药。口服给药时,它表现出良好的肠道吸收。肾小球滤过后,ALA在肾近端小管中也能有效重吸收。然而,其跨膜转运进入肠和肾上皮细胞的途径和机制尚不清楚。在这里,我们证明ALA利用肠和肾顶端肽转运体进入上皮细胞。在表达克隆的肠肽转运体PEPT1或肾型PEPT2的非洲爪蟾卵母细胞和毕赤酵母细胞中测定了ALA转运的动力学和特征。通过在这些异源表达系统中使用放射性标记的ALA和电生理技术,我们确定:(a)PEPT1和PEPT2通过可饱和的、pH依赖性的转运机制转运3H-ALA;(b)ALA和二肽/三肽,而不是GABA或相关氨基酸,在载体的同一底物结合位点竞争;(c)由于H+/ALA共转运,ALA转运本质上是生电性的。用兔组织中PEPT1和PEPT2的特异性引物进行的逆转录酶-PCR分析表明,特别是PEPT2 mRNA在包括肺、脑和乳腺在内的多种其他组织中表达,这些组织已被证明能积累ALA。这表明这些组织可能通过表达的肽转运体摄取卟啉前体,为有效的光动力疗法提供内源性光敏剂。