鹿茸水溶性蛋白的结构表征与抗疲劳活性研究

Structural characterization and anti-fatigue activity of water-soluble protein of velvet antler

目的:探索鹿茸水溶性蛋白的结构特征,分析其抗疲劳的作用功效。方法:以梅花鹿鹿茸为原料,通过碱提酸沉法提取鹿茸水溶性蛋白,通过SDS-PAGE凝胶电泳结合考马斯亮蓝染色法检测其相对分子质量分布,通过高效液相色谱法分析其氨基酸组成,采用紫外光谱、圆二色光谱、红外光谱、内源荧光光谱等光谱学方法,以及扫描电子显微镜检测其结构特征和微观分子形态,利用大鼠力竭游泳实验评价其抗疲劳活性。结果:鹿茸水溶性蛋白的主要相对分子质量分布为(43~80)×10^(3)及<14.4×10^(3),其中含有丰富的谷氨酸、赖氨酸、天冬氨酸和亮氨酸,以及丰富的必需氨基酸。鹿茸水溶性蛋白具有明显的蛋白质特征结构,包含丰富的α-螺旋结构,具有良好的蛋白稳定性,微观形态多呈片状和无规则树枝状。其能够明显延长大鼠的力竭游泳时间,降低血液中尿素氮和乳酸的含量,增加肝糖原和肌糖原的累积,提高乳酸脱氢酶(lactate dehydrogenase,LDH)和超氧化物歧化酶(superoxide dismutase,SOD)的活性,降低丙二醛(malondialdehyde,MDA)的含量。结论:鹿茸水溶性蛋白的分子特征可能是其具有良好抗疲劳活性的结构基础。

OBJECTIVE To explore the structural characteristics of water-soluble protein of velvet antler and examine its anti-fatigue effect.METHODS Water-soluble protein of velvet antler was extracted with sika deer antler as raw material through alkaline extraction and acid precipitation.Molecular weight distribution was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Coomassie blue stain.Amino acid composition was detected by high performance liquid chromatography(HPLC).And structural characteristics and micro-molecular morphologies were detected by ultraviolet(UV) spectroscopy,circular dichroism(CD) spectroscopy,Fourier transform infrared spectroscopy(FTIR),endogenous fluorescent spectroscopy and scanning electron microscopy(SEM).Anti-fatigue activity was evaluated by exhaustive swimming experiments in mice.RESULTS Molecular weight distribution of water-soluble protein ranged from(43-80)×10^(3) and it was less than 14.4×10^(3).It contained an abundant supply of glutamic acid,lysine,aspartic acid and leucine,as well as essential amino acids.Watersoluble protein of velvet antler possessed distinct protein characteristic structures,including abundant alpha helix and excellent protein stabilities.It prolonged exhausted swimming time,reduced the blood levels of urea nitrogen and lactated,caused an accumulation of liver glycogen and muscle glycogen,boosted the activities of lactate dehydrogenase(LDH) and superoxide dismutase(SOD) and lowered the content of malondialdehyde(MDA).CONCLUSION The molecular characteristics of water-soluble proteins of velvet antler may serve as a structural basis for their excellent anti-fatigue properties.