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兔髓袢升支粗段外髓部来源的肾细胞中胆碱转运及其渗透调节

Choline transport and its osmotic regulation in renal cells derived from the rabbit outer medullary thick ascending limb of Henle.

作者信息

Grunewald R W, Oppermann M, Müller G A

机构信息

Department of Nephrology and Rheumatology, University of Göttingen, Robert-Koch-Strasse 40, D-37075 Göttingen, Germany.

出版信息

Pflugers Arch. 1997 Nov;434(6):815-21. doi: 10.1007/s004240050470.

Abstract

Organic osmolytes such as betaine and glycerophosphorylcholine (GPC) are of major importance concerning volume regulation of inner and outer medullary epithelial cells. Recently we demonstrated that the intracellular betaine content in rabbit kidney cells derived from the outer medullary thick ascending limb of Henle's loop (TALH) is osmotically regulated by betaine synthesis. In this context it was our purpose to characterize the uptake of choline, a precursor of betaine and GPC. We found TALH cells to possess a specific choline transport system with a maximum velocity (Vmax) of 71 +/- 12 pmol . micro l-1 cell water . min-1 and an apparent affinity (Km) of 155 +/- 19 micromol . l-1. The uptake of choline was sodium independent and not electrogenic, but it was significantly reduced by the removement of chloride from the incubation medium. After long-term adaptation of TALH cells to a hyperosmotic medium (600 mosmol . l-1, osmolarity adjusted with NaCl or urea) a significant higher choline uptake rate was observed (Vmax: 166 +/- 9 (NaCl), 96 +/- 12 (urea) pmol . microl-1 cell water . min-1). Our results suggest that the uptake of choline is due to higher intracellular requirements of choline under hypertonic conditions. Finally, an increase in the Vmax of the choline transport system may enable sufficient synthesis of betaine and GPC.

摘要

诸如甜菜碱和甘油磷酰胆碱(GPC)等有机渗透物对于肾内、外髓质上皮细胞的容积调节至关重要。最近我们证明,源自亨氏袢外髓质厚升支(TALH)的兔肾细胞内的甜菜碱含量受甜菜碱合成的渗透调节。在此背景下,我们的目的是表征甜菜碱和GPC的前体胆碱的摄取情况。我们发现TALH细胞具有一种特定的胆碱转运系统,其最大速度(Vmax)为71±12 pmol·μl⁻¹细胞水·min⁻¹,表观亲和力(Km)为155±19 μmol·L⁻¹。胆碱的摄取不依赖于钠且不产生电效应,但通过从孵育培养基中去除氯离子可使其显著降低。在将TALH细胞长期适应高渗培养基(600 mosmol·L⁻¹,用NaCl或尿素调节渗透压)后,观察到胆碱摄取率显著更高(Vmax:166±9(NaCl),96±12(尿素)pmol·μl⁻¹细胞水·min⁻¹)。我们的结果表明,胆碱摄取是由于高渗条件下细胞内对胆碱的需求增加。最后,胆碱转运系统Vmax的增加可能使甜菜碱和GPC能够充分合成。

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