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大鼠肾基底外侧膜对长链脂肪酸的介导转运。

Mediated transport of long-chain fatty acids by rat renal basolateral membranes.

作者信息

Trimble M E

机构信息

Department of Physiology, State University of New York Health Science Center, Syracuse 13210.

出版信息

Am J Physiol. 1989 Oct;257(4 Pt 2):F539-46. doi: 10.1152/ajprenal.1989.257.4.F539.

Abstract

Transport of long-chain fatty acids (LCFA) was studied in isolated rat renal cortical membrane vesicles to determine the mechanism of entry of these metabolic substrates into renal cells. Total uptake of palmitate into basolateral (BLMV) and brush-border membrane vesicles (BBMV) was determined with the use of a rapid filtration technique. In the presence of an inwardly directed H+ gradient (pHo = 6.0, pHi = 7.4) palmitate (0.26 microM) uptake went through a maximum ("overshoot") in BLMV but not in BBMV. The amount of palmitate nonspecifically bound to vesicles was subtracted from total uptake to determine intravesicular content. Peak values for intravesicular palmitate content were 26 +/- 9 pmol/mg protein, 2.5-fold higher than 60-min values of 10 +/- 2 pmol/mg (P less than 0.05, n = 6). The overshoot could not be accounted for by changes in voltage. An inwardly directed Na+ gradient was without effect on palmitate uptake in BLMV and BBMV. pH gradient-stimulated palmitate transport (15 s) in BLMV was subject to cis-inhibition by 4,4'-diisothiocyanatostilbene-,,2-disulfonic acid. Probenecid and p-aminohippurate efflux from BLMV resulted in counter-flow accumulation of palmitate. It is concluded that at least a portion of LCFA transport across the basolateral membrane is mediated and may occur via anion exchange.

摘要

为了确定这些代谢底物进入肾细胞的机制,我们在分离的大鼠肾皮质膜囊泡中研究了长链脂肪酸(LCFA)的转运。使用快速过滤技术测定了棕榈酸酯进入基底外侧(BLMV)和刷状缘膜囊泡(BBMV)的总摄取量。在存在内向H⁺梯度(pHo = 6.0,pHi = 7.4)的情况下,棕榈酸酯(0.26微摩尔)在BLMV中的摄取出现最大值(“过冲”),而在BBMV中则没有。从总摄取量中减去非特异性结合到囊泡上的棕榈酸酯量,以确定囊泡内含量。囊泡内棕榈酸酯含量的峰值为26±9皮摩尔/毫克蛋白质,比60分钟时10±2皮摩尔/毫克的值高2.5倍(P<0.05,n = 6)。过冲不能用电压变化来解释。内向的Na⁺梯度对BLMV和BBMV中棕榈酸酯的摄取没有影响。pH梯度刺激的BLMV中棕榈酸酯转运(15秒)受到4,4'-二异硫氰基芪-2,2-二磺酸的顺式抑制。丙磺舒和对氨基马尿酸从BLMV流出导致棕榈酸酯的逆流积累。结论是,至少一部分LCFA跨基底外侧膜的转运是介导的,可能通过阴离子交换发生。

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