重水标记法测定大鼠骨骼肌蛋白合成动力学相关参数

发布时间：2018-03-30 09:52

本文选题：重水　切入点：骨骼肌　出处：《生理学报》2017年03期

【摘要】：本研究旨在以重水(~2H_2O)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~2H_2O,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2~H-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f_t=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。
[Abstract]:The purpose of this study was to determine the parameters related to the synthesis kinetics of skeletal muscle protein in rats using 2H _ 2O as tracer. Twenty Sprague Dawley Dawley SD rats were injected intraperitoneally and labeled with 2H _ 2O in the form of daily drinking water. Plasma was extracted from each of 4 rats by heart puncture, and quadriceps femoris was isolated. Skeletal muscle protein and plasma free amino acids were extracted and hydrolyzed with hydrochloric acid. After derivation, the abundance of 2H-alanine in skeletal muscle protein and free alanine in plasma were detected by gas chromatography-mass spectrometer (chromatography-mass). The fractional synthesis rate and synthesis kinetics equation of skeletal muscle protein were calculated. The fraction synthesis rate of skeletal muscle protein was 12.8g / week, and the synthesis kinetics equation was f_t=0.158 脳 1-eosin -0.128tl. These results suggest that the heavy water labeling method can effectively determine the synthesis kinetic parameters of skeletal muscle protein, and it is suitable for long-term labeling experiment.
【作者单位】：扬州大学旅游烹饪学院营养系;
【基金】：supported by the National Natural Science Foundation of China(No.81472963)
【分类号】：R68

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