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通过改变细胞膜流动性调节肾脏硫酸盐转运

Modulation of sulfate renal transport by alterations in cell membrane fluidity.

作者信息

Lee H J, Balasubramanian S V, Murer H, Biber J, Morris M E

机构信息

Department of Pharmaceutics, School of Pharmacy, State University of New York at Buffalo, Amherst, New York 14260, USA.

出版信息

J Pharm Sci. 1999 Oct;88(10):976-80. doi: 10.1021/js990114c.

Abstract

Changes in membrane fluidity have been shown to alter the sodium-dependent renal transport of glucose and phosphate; however, this has not been examined for sodium/sulfate cotransport in the renal proximal tubule. Sodium/sulfate cotransport regulates the homeostasis of sulfate in mammals. The objective of this study was to investigate the influence of alterations of membrane fluidity on sodium-coupled sulfate transport in the Madin-Darby canine kidney cells, which have been stably transfected with sodium/sulfate cotransporter (NaSi-1) cDNA (MDCK-Si). Preincubation of cells with 0. 2 mM cholesterol significantly decreased the V(max) for sodium/sulfate cotransport (13.69 +/- 1.11 vs 10.15 +/- 1.17 nmol/mg protein/5 min, mean +/- SD, n = 4, p < 0.01) with no significant alteration in K(m). The addition of benzyl alcohol (20 mM) to cells increased the V(max) of sulfate uptake by 20% (11.97 +/- 0.91 vs 14. 35 +/- 0.56 nmol/mg protein/5 min, mean +/- SD, n = 3, p < 0.05) with no significant change in K(m). Membrane fluidity, as measured by the fluorescence polarization of 1,6-diphenyl 1,3,5-hexatriene (DPH), was significantly increased in MDCK-Si cells treated with 20 mM benzyl alcohol and decreased in the cells preincubated with 0.2 mM cholesterol, compared with control cells. Our results suggest that alterations in membrane fluidity that may occur as a result of disease states, aging, and pregnancy may play an important role in the modulation of renal sodium/sulfate cotransport.

摘要

膜流动性的改变已被证明会改变钠依赖性肾对葡萄糖和磷酸盐的转运;然而,肾近端小管中钠/硫酸盐协同转运的情况尚未得到研究。钠/硫酸盐协同转运调节哺乳动物体内硫酸盐的稳态。本研究的目的是探讨膜流动性改变对稳定转染了钠/硫酸盐协同转运体(NaSi-1)cDNA的Madin-Darby犬肾细胞(MDCK-Si)中钠偶联硫酸盐转运的影响。用0.2 mM胆固醇预孵育细胞可显著降低钠/硫酸盐协同转运的V(max)(13.69±1.11对10.15±1.17 nmol/mg蛋白/5分钟,平均值±标准差,n = 4,p < 0.01),而K(m)无显著改变。向细胞中添加苄醇(20 mM)可使硫酸盐摄取的V(max)增加20%(11.97±0.91对14.35±0.56 nmol/mg蛋白/5分钟,平均值±标准差,n = 3,p < 0.05),K(m)无显著变化。与对照细胞相比,用20 mM苄醇处理的MDCK-Si细胞中,通过1,6-二苯基-1,3,5-己三烯(DPH)荧光偏振测量的膜流动性显著增加,而用0.2 mM胆固醇预孵育的细胞中膜流动性降低。我们的结果表明,疾病状态、衰老和妊娠可能导致的膜流动性改变可能在调节肾钠/硫酸盐协同转运中起重要作用。

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