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重水标记法测定大鼠骨骼肌蛋白合成动力学相关参数

田颖, 戴倩倩, 孟婵芳, 孙翌, 彭景, 陈玉

扬州大学旅游烹饪学院营养系，扬州 225127

摘要

本研究旨在以重水(2H2O)为示踪剂，测定大鼠骨骼肌蛋白合成动力学相关参数。将20只Sprague Dawley (SD)大鼠以腹腔注射、日常饮水的形式标记2H2O，分别在首次标记后的第1、3、5、6和10周，各取4只大鼠心脏穿刺取血浆，分离股四头肌。提取骨骼肌蛋白和血浆游离氨基酸，用盐酸水解，衍生后用气相色谱-质谱仪(gas chromatography-mass spectrometry, GC-MS)检测骨骼肌蛋白2H-丙氨酰基和血浆游离丙氨酸中2H的丰度，计算骨骼肌蛋白的分数合成率、合成动力学方程。结果显示，骨骼肌蛋白的分数合成率为12.8%/周，合成动力学方程为ft = 0.158 × (1 − e−0.128t)。以上结果提示，重水标记法能有效测定骨骼肌蛋白的合成动力学参数，适用于长期标记实验。

关键词： 重水; 骨骼肌; 分数合成率

分类号：R33-3

[Measurement of kinetic parameters of skeletal muscle protein synthesis in rats by deuterated water.] [Article in Chinese]

TIAN Ying, DAI Qian-Qian, MENG Chan-Fang, SUN Yi, PENG Jing, CHEN Yu

Department of Nutrition, School of Tourism and Culinary Science, Yangzhou University, Yangzhou 225127, China

Abstract

The aim of the present study was to measure the kinetic parameters of skeletal muscle protein synthesis in rats by deuterated water (2H2O). Twenty Sprague-Dawley (SD) rats were labeled by 2H2O through intraperitoneal injection and drinking. At the each end of the 1st, 3rd, 5th, 6th and 10th week after the first 2H2O labeling, four rats were sacrificed by cardiac puncture for blood plasma and quadriceps femoris sampling. Skeletal muscle protein and free amino acids in plasma were purified, hydrolyzed by hydrochloric acid and derived. The deuterium enrichments of 2H-labeled alanyl in skeletal muscle protein and plasma protein-bound 2H-labeled alanine were determined by gas chromatography-mass spectrometry (GC-MS). The fractional synthesis rate of skeletal muscle protein and synthetic dynamic equation were calculated. The fractional synthetic rate of skeletal muscle protein was 12.8%/week, and synthetic dynamic equation was ft = 0.158 × (1 − e−0.128t). The results suggest that the kinetic parameters of skeletal muscle protein synthesis can be measured by 2H2O labeling, and the method can be applied in long-term labeling experiment.

Key words: deuterated water; skeletal muscle; fractional synthesis rate

收稿日期：2016-11-08　　录用日期：2016-12-31

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引用本文：

田颖, 戴倩倩, 孟婵芳, 孙翌, 彭景, 陈玉. 重水标记法测定大鼠骨骼肌蛋白合成动力学相关参数[J]. 生理学报 2017; 69 (3): 311-315.

TIAN Ying, DAI Qian-Qian, MENG Chan-Fang, SUN Yi, PENG Jing, CHEN Yu. [Measurement of kinetic parameters of skeletal muscle protein synthesis in rats by deuterated water.] [Article in Chinese]. Acta Physiol Sin 2017; 69 (3): 311-315 (in Chinese with English abstract).