| 李世超1， 刘诗超2， 赵大庆1， 王思明1， 睢博文1.火麻仁蛋白酶解工艺及其活性肽对骨骼肌细胞
损伤保护作用机制研究[J].中国油脂,2026,52(3):.[LI Shichao1, LIU Shichao2 , ZHAO Daqing1, WANG Siming1, SUI Bowen1.Enzymatic hydrolysis process of hemp seed protein and the protective mechanism of its active peptide against skeletal muscle cell injury[J].China Oils and Fats,2026,52(3):.] |
| 火麻仁蛋白酶解工艺及其活性肽对骨骼肌细胞
损伤保护作用机制研究 |
| Enzymatic hydrolysis process of hemp seed protein and the protective mechanism of its active peptide against skeletal muscle cell injury |
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投稿时间：2024-11-29 修订日期：2025-09-04 录用日期：2025-02-24
出版日期：2026-03-20
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| DOI：10.19902/j.cnki.zgyz.1003-7969.240670 |
| 中文关键词: 火麻仁活性肽 酶解工艺 氧化应激 骨骼肌细胞 |
| 英文关键词:hemp seed active peptide enzymatic process oxidative stress skeletal muscle cell |
| 基金项目:吉林省重大科技专项（20210304002YY） |
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| Author Name | Affiliation | | LI Shichao1, LIU Shichao2 , ZHAO Daqing1, WANG Siming1, SUI Bowen1 | 1.Northeast Asia Institute of Traditional Chinese Medicine, Changchun University of Traditional
Chinese Medicine, Changchun 130117, China； 2.School of Pharmacy, Changchun
University of Traditional Chinese Medicine, Changchun 130117, China |
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| 中文摘要: |
| 为了有效提高火麻仁饼粕的利用率，以火麻仁蛋白（HPI）为原料，采用酶解工艺制备火麻仁活性肽（HP），并探讨其对骨骼肌细胞损伤的保护作用及其机制。采用H2O2诱导的体外氧化应激损伤模型，以含200 μg/mL HP的培养基培养小鼠成肌细胞（C2C12细胞），以细胞存活率为评价指标，筛选蛋白酶，并利用单因素实验结合正交实验优化酶解工艺。对HP进行蛋白电泳分析、氨基酸分析，并考察HP对细胞氧化应激指标和炎症因子水平、细胞分化及Ca2+摄取能力的影响。结果表明：HPI的最优酶解工艺为分步酶解，即在pH 7.0、酶解温度50 ℃、中性蛋白酶添加量1.5%的条件下酶解8 h，再在pH 6.5、酶解温度50 ℃、风味酶添加量2%的条件下酶解6 h，在此条件下细胞存活率为90.56%；蛋白电泳结果显示，HP的分子质量较HPI显著降低；氨基酸分析结果显示，HP中17种氨基酸含量均高于HPI；经HP处理后，白细胞介素6（IL-6）、白细胞介素1β(IL-1β)和肿瘤坏死因子α（TNF-α）水平显著降低(p<0.01)，MDA含量、LDH活力均显著降低(p<0.05，p<0.01)，SOD、CAT活力均显著升高(p<0.01)，细胞分化和Ca2+摄取能力显著增强。综上，以中性蛋白酶和风味酶分步酶解可以得到对骨骼肌细胞损伤具有保护作用的HP，其可通过增强氧化损伤的骨骼肌细胞Ca2+的摄取能力来发挥保护作用。 |
| 英文摘要: |
| In order to effectively improve the utilization rate of hemp seed cake and meal, hemp seed protein (HPI) was used as raw material to prepare hemp seed active peptide (HP) by enzymatic hydrolysis process, and its protective effect on skeletal muscle cell injury and its mechanism were discussed. The oxidative stress injury model induced by H2O2 in vitro was used, and mouse myoblasts (C2C12 cells) was cultured with medium containing 200 μg/mL HP. The cell viability was used as the evaluation index to screen proteases, and the enzymatic hydrolysis process was optimized by single factor experiment combined with orthogonal experiment. Protein electrophoresis analysis and amino acid analysis were performed on HP, and the effects of HP on cell oxidative stress indicators and inflammatory factor levels, cell differentiation and Ca2+ uptake capacity were investigated. The results showed that the optimum enzymatic hydrolysis process of HPI was stepwise enzymatic hydrolysis. Under the conditions of pH 7.0, enzymatic hydrolysis temperature 50 ℃, neutral protease dosage 1.5%, enzymatic hydrolysis time 8 h, and pH 6.5, enzymatic hydrolysis temperature 50 ℃, flavor enzyme dosage 2%, enzymatic hydrolysis time 6 h, the cell viability was 90.56%. The results of protein electrophoresis showed that the molecular weight of HP was significantly lower than that of HPI. The results of amino acid composition analysis showed that the contents of 17 amino acids in HP were higher than those in HPI.After HP treatment, the levels of IL-6, IL-1β and TNF-α significantly decreased (p<0.01), the content of MDA and the activity of LDH significantly decreased (p<0.05, p<0.01), the activities of SOD and CAT significantly increased (p<0.01), and the cell differentiation and Ca2+ uptake capacity were significantly enhanced. In summary, HP with protective effect on skeletal muscle cell injury can be obtained by stepwise enzymatic hydrolysis with neutral protease and flavor enzyme, which can play a protective role by enhancing the uptake of Ca2+ in oxidatively damaged skeletal muscle cells. |
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