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大鼠肺II型上皮细胞对钠依赖性和非依赖性胆碱的摄取。

Sodium-dependent and -independent choline uptake by type II epithelial cells from rat lung.

作者信息

van Rossum G D, Boyd C A

机构信息

Department of Pharmacology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.

出版信息

J Membr Biol. 1998 Mar 15;162(2):147-56. doi: 10.1007/s002329900352.

Abstract

The uptake of 3H-labeled choline by a suspension of isolated type II epithelial cells from rat lung has been studied in a Ringer medium. Uptake was linear for 4 min at both 0.1 microM and 5.0 microM medium choline; at 5 microM, only 10% of the label was recovered in a lipid fraction. Further experiments were conducted at the low concentration (0.1 microM), permitting characterization of the properties of high-affinity systems. Three fractions of choline uptake were detected: (i) a sodium-dependent system that was totally inhibited by hemicholinium-3 (HC-3); (ii) a sodium-independent uptake, when Na+ was replaced by Li+, K+ or Mg2+, inhibited by HC-3; (iii) a residual portion persisting in the absence of Na+ and unaffected by HC-3. Choline uptake was sigmoidally related to the medium Na+ concentration. Kinetic properties of the uptake of 0.1 microM 3H-choline in the presence and absence of medium Na+ were examined in two ways. (a) Inhibition by increasing concentrations of unlabeled choline (0.5-100 microM) was consistent with the presence of two Michaelis-Menten-type systems in the presence of Na+; a Na(+)-dependent portion (a mean of 0.52 of the total) had a K(m) for choline of 1.5 microM while K(m) in the absence of Na+ (Li+ substituting) was 18.6 microM. (b) Inhibition by HC-3 (0.3-300 microM) gave Ki values of 1.7 microM and 5.0 microM HC-3 for the Na(+)-dependent and -independent fractions. The apparent K(m) of the Na(+)-dependent uptake is lower than that reported previously for lung-derived cells and is in the range of the K(m) values reported for high-affinity, Na(+)-dependent choline uptake by neuronal cells.

摘要

在林格氏液中,对从大鼠肺中分离出的II型上皮细胞悬液摄取3H标记胆碱的情况进行了研究。在培养基胆碱浓度为0.1微摩尔/升和5.0微摩尔/升时,摄取在4分钟内呈线性;在5微摩尔/升时,仅有10%的标记物在脂质部分中被回收。在低浓度(0.1微摩尔/升)下进行了进一步实验,以表征高亲和力系统的特性。检测到胆碱摄取的三个部分:(i)一种依赖钠的系统,被半胱氨酸-3(HC-3)完全抑制;(ii)一种不依赖钠的摄取,当Na+被Li+、K+或Mg2+取代时,被HC-3抑制;(iii)在无Na+时持续存在且不受HC-3影响的残余部分。胆碱摄取与培养基Na+浓度呈S形关系。通过两种方式研究了在有和无培养基Na+存在时,0.1微摩尔/升3H-胆碱摄取的动力学特性。(a)用增加浓度的未标记胆碱(0.5 - 100微摩尔/升)进行抑制,这与在有Na+存在时存在两个米氏类型系统一致;一个依赖Na+的部分(占总量的平均值为0.52)对胆碱的K(m)为1.5微摩尔/升,而在无Na+(Li+替代)时K(m)为18.6微摩尔/升。(b)用HC-3(0.3 - 300微摩尔/升)进行抑制,对于依赖Na+和不依赖Na+的部分,HC-3的Ki值分别为1.7微摩尔/升和5.0微摩尔/升。依赖Na+摄取的表观K(m)低于先前报道的肺源性细胞的值,且在神经元细胞高亲和力、依赖Na+的胆碱摄取所报道的K(m)值范围内。

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