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大鼠单根近端肾小管Na~(+)--K~(+)--ATPase活性测定方法的改进

高原, 罗蕾, 刘红

遵义医学院珠海校区生理学教研室.广东,珠海 519041

摘要

该研究旨在对Doucet等报道的定量检测大鼠单根近端肾小管Na~(+)--K~(+)--ATPase活性方法进行改进。取经过Ⅱ型胶原酶消化的大鼠肾脏皮质组织，在体视显微镜下手工分离单根近端肾小管，并测量其长度，经低渗和冻融处理后与[#gamma#--~(32)P]ATP共同孵育，液闪法检测从[#gamma#--~(32)P]ATP解离出的~(32)Pi，采用修正后的公式计算Na~(+)--K~(+)--ATPase活性。改良法与Doucet等的方法比较，测定单根近端肾小管Na~(+)--K~(+)--ATPase活性无显著性差异（{sl P}>0.05）。改进后的方法节省试剂，操作简便、省时。

关键词： 近端肾小管; Na~(+)-K~(+)-ATPase; 大鼠

An improved approach for determination of Na~(+)--K~(+)--ATPase activity in single proximal renal tubule of rat

Gao Yuan, Luo Lei, Liu Hong

Department of Physiology, Zunyi Medical College Zhuhai Campus.Zhuhai 519041,Guangdong

Abstract

An improved approach for determination of Na~(+)-K~(+)-ATPase activity in single proximal renal tubule of rat reported by Doucet and his colleagues has been suggested. The single proximal renal tubules were isolated by hand under stereomicroscope from collagenase Ⅱ-treated renal cortical tissue in rats. The length of every single renal tubule segment obtained was measured. The single proximal renal tubules were treated with a hypoosmotic solution and with freeze-thaw successively, before incubation with [#gamma#-~(32)P]ATP. ~(32)Pi hydrolyzed from [#gamma#-~(32)P]ATP by Ⅱ in the single proximal renal tubules was assayed by liquid scintillation counting. The activity of Na~(+)-K~(+)-ATPase in the single proximal renal tubules was calculated by applying a modified formula. There was no significant difference in the measurement result of Na~(+)-K~(+)-ATPase activity between the method of Doucet et al. and the improved one, but the latter has advantage of less time, less reagents and being easy to operate.

Key words: proximal renal tubule;Na~(+)-K~(+)-ATPase;rat

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引用本文：

高原, 罗蕾, 刘红. 大鼠单根近端肾小管Na~(+)--K~(+)--ATPase活性测定方法的改进[J]. 生理学报 2007; 59 (3): 382-386.

Gao Yuan, Luo Lei, Liu Hong. An improved approach for determination of Na~(+)--K~(+)--ATPase activity in single proximal renal tubule of rat. Acta Physiol Sin 2007; 59 (3): 382-386 (in Chinese with English abstract).