小牛胸腺肽提取工艺优化及其抗疲劳活性研究  

Study of Optimization of Extraction Process and Anti-fatigue Activity of Calf Thymus Polypeptide

作　　者：赵辰菲 谭云云 李森多 林子涵 刘占峰 李兰洲 ZHAO Chenfei;TAN Yunyun;LI Senduo;LIN Zihan;LIU Zhanfeng;LI Lanzhou(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学食药用菌教育部工程研究中心,吉林长春130118

出　　处：《药物生物技术》2024年第6期636-642,共7页Pharmaceutical Biotechnology

摘　　要：通过对小牛胸腺肽提取工艺优化,从而提高提取率、提升品质,并在明确小牛胸腺肽物质基础上对其抗疲劳活性进行评价,以期为口服类胸腺肽药物质量标准提升及临床应用的选择提供研究基础。通过单因素实验法对胸腺肽提取工艺中的酶解时间、酶解温度等条件进行优化,并选用凝胶渗透色谱法和高效液相色谱法对其分子量范围和氨基酸组成含量进行分析。在Balb/c小鼠中,通过疲劳转棒实验、耐力跑实验等行为学实验评价胸腺肽抗疲劳活性,通过测定乳酸、乳酸脱氢酶等生化指标,对胸腺肽的抗疲劳活性机制进行探索。胸腺肽中很大部分是小分子组分,相对分子质量在800以下的组分占53.42%,600以下组分占30.82%,谷氨酸(8.40 g/kg)、赖氨酸(7.96 g/kg)和脯氨酸(13.24 g/kg)是含量最多的三种氨基酸残基。并证实胸腺肽具有显著的抗疲劳调节机体状态的作用,其机制与降低机体的乳酸和氧化应激相关。本研究优化了一种小牛胸腺肽的提取工艺,明确了其物质组成,并证实小牛胸腺肽具有良好的抗疲劳活性,可能有助于机体状态的恢复,提升患者治疗耐受能力。By optimizing the extraction process of calf thymic polypeptide,the extraction rate and quality were improved,and the anti-fatigue activity of calf thymic peptide was evaluated on the basis of identifying the substance of calf thymic peptide,in order to provide a research basis for the improvement of the quality standard of oral thymosin-like drugs and the selection of clinical application.The enzymolysis time and enzymolysis temperature in the extraction process of calf thymic polypeptide were optimized by single factor experiment.The molecular weight range and amino acid composition of calf thymic polypeptide were analyzed by gel permeation chromatography and high performance liquid chromatography.In Balb/c mice,the anti-fatigue activity of calf thymic polypeptide was evaluated by fatigue rotating test,endurance running test,as well as determination of the content of lactic acid(LD),lactate dehydrogenase(LDH),reactive oxygen species(ROS)and superoxide dismutase(SOD)to explore the mechanism of anti-fatigue activity of calf thymic polypeptide.Most thymosins were small molecular components,with the molecular weight below 800 dalton accounting for 53.42%,and the molecular weight below 600 dalton accounting for 30.82%.Clutamate(8.40 g/kg),lysine(7.96 g/kg)and proline(13.24 g/kg)were the most abundant amino acid residues of calf thymic polypeptide.It was confirmed in mice that calf thymus polypeptide has a good effect of anti-fatigue and regulating the body state,and its mechanism is related to the reduction of LD and oxidative stress.This study optimized the extraction process of calf thymus polypeptide,clarified the material composition of calf thymus polypeptide,and confirmed that calf thymus peptide has good anti-fatigue activity,which may contribute to the recovery of body state and improve the treatment tolerance of patients.

关 键 词：胸腺肽 工艺优化 物质组成 抗疲劳 乳酸 氧化应激 

分 类 号：R963[医药卫生—微生物与生化药学]