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原代培养的II型肺上皮细胞对亚精胺的摄取。

Spermidine uptake by type II pulmonary epithelial cells in primary culture.

作者信息

Kameji R, Rannels S R, Pegg A E, Rannels D E

机构信息

Department of Physiology, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.

出版信息

Am J Physiol. 1989 Jan;256(1 Pt 1):C160-7. doi: 10.1152/ajpcell.1989.256.1.C160.

Abstract

The transport pathway for the polyamine spermidine (SPD) was characterized in primary isolates of type II pulmonary epithelial cells from rat lungs. [14C]spermidine was accumulated by type II cells via a temperature-, sodium-, and concentration-dependent saturable pathway, with an apparent Km of 0.48 microM and a maximum velocity (V max) of 0.32 pmol.microgram DNA-1.min-1. SPD uptake was inhibited by gramicidin and by reduced extracellular sodium but was unaffected by alpha-aminoisobutyric acid (AIB), which entered the cells by a similar saturable pathway. Uptake of SPD also was inhibited by the exogenous polyamines putrescine (PUTR) and spermine (SPM), as well as by the methylglyoxal bis(guanylhydrazone) (MGBG) and by paraquat (PQ). The order of potency of these inhibitors was SPM greater than PUTR = MGBG much greater than PQ. The absence of serum reduced the Vmax of the system slightly but had no effect on the apparent Km. In contrast, after 3 days in primary cell culture, the kinetics of SPD transport were altered by decreases in both the Km and Vmax of the uptake process. These observations indicate that type II pulmonary epithelial cells exhibit a pathway of polyamine uptake with general characteristics similar to those observed previously in intact lung tissue and other cell types.

摘要

在大鼠肺脏II型肺上皮细胞的原代分离物中对多胺亚精胺(SPD)的转运途径进行了表征。[14C]亚精胺通过一种温度、钠和浓度依赖性的可饱和途径被II型细胞积累,其表观Km为0.48微摩尔,最大速度(Vmax)为0.32皮摩尔·微克DNA-1·分钟-1。短杆菌肽和细胞外钠浓度降低可抑制SPD的摄取,但α-氨基异丁酸(AIB)对其无影响,AIB通过类似的可饱和途径进入细胞。SPD的摄取也受到外源性多胺腐胺(PUTR)和精胺(SPM)以及甲基乙二醛双(胍腙)(MGBG)和百草枯(PQ)的抑制。这些抑制剂的效力顺序为SPM大于PUTR = MGBG远大于PQ。血清的缺乏使该系统的Vmax略有降低,但对表观Km无影响。相反,在原代细胞培养3天后,SPD转运的动力学因摄取过程的Km和Vmax均降低而发生改变。这些观察结果表明,II型肺上皮细胞表现出一种多胺摄取途径,其一般特征与先前在完整肺组织和其他细胞类型中观察到的相似。

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