胎鼠、成鼠和人胚的骨骼肌提取液及肌源神经营养因子的高效液相分析

THE HPLC ANALYSIS OF SKELETAL MUSCLE EXTRACT AND MUSCLE DERIVED NEUROTROPHIC FACTOR FROM EMBRYONIC AND ADULT RAT AND HUMAN EMBRYO

大量的在体和离体实验证明，骨骼肌内存在着促进脊髓神经元存活和再生的物质。本研究采用高效液相分析方法对胎鼠、成鼠和人胚的骨骼肌提取液进行了分析。结果表明，三者在相同条件下尽管出现蛋白峰的数量不同，峰值也有所差异，但在相同的时间点出现两处４ 峰（Ⅰ、Ⅱ１、Ⅱ２、Ⅱ３）的基本波型。峰Ⅱ１ 的分子量为３５０００Ｄ，峰Ⅱ２ 的分子量为２２０００Ｄ，峰Ⅱ３ 的分子量为１８０００Ｄ。胎鼠和人胚的三峰呈中、低、高的形态分布，而成鼠的三峰呈低、中、高的形态分布。肌源神经营养因子（１０～３０ ｋＤ）位于峰Ⅱ。此结果使人们对肌源神经营养因子的存在产生了疑问，肌源神经营养因子究竟是一个新的营养因子（分子量究竟是多少？），还是已知的神经营养因子（ＣＮＴＦ，ＢＤＮＦ，ＮＴ３ 等）在骨骼肌组织中的分布？ 对此有必要进行分子生物学的研究来确定。

A series of studies in vivo and in vitro have demonstrated previously that skeletal muscle contains a substance which selectively enhances the survival and regeneration of spinal cord neurons. In the present study, by using the HPLC analysis showed that an active fraction, skeletal muscle derived neurotrophic factor(10～30 kD)was localized. Also under the same analysis, though the number and the value of the protein's peaks were different in three kinds of skeletal muscle extracts(adult rat, embryonic rat and human embryo), they showed nearly the same peak wave line charts. Four peaks were divided into two groups(Ⅰ、Ⅱ 1、Ⅱ 2、Ⅱ 3). The molecular weight of peak Ⅱ 1 is about 35 kD, peak Ⅱ 2 is 22 kD, and peak Ⅱ 3 is 18 kD. The muscle derived neurotrophic factor(10～30 kD) is localized in peak Ⅱ group. From the result there is some doubt whether the muscle derived neurotrophic factor is a new factor(What is exactly it's molecular weight?) or the identified factors(such as CNTF, BDNF, NT 3) were contained by skeletal muscle? Further molecular biological study is necessary.