海带活性物质对秀丽隐杆线虫的降脂作用

谢幸媛; 李娜; 汪鹏翼; 刘斌; 曾峰

doi:10.13386/j.issn1002-0306.2024120196

海带活性物质对秀丽隐杆线虫的降脂作用

Hypolipidemic Effects of Active Substances from Laminaria japonica on Caenorhabditis elegans

摘要

摘要: 为制备海带（Laminaria japonica，L. japonica）活性物质并分析其物质组成和降脂活性。利用不同极性的溶剂结合超声辅助醇提取、水提取、复合蛋白酶酶解以及碳酸钠提取分别制备海带醇溶性物质（alcohol extract of Laminaria japonica，LJA）、海带水溶性物质（water extract of Laminaria japonica，LJW）、海带酶解物质（enzymes hydrolyzed of Laminaria japonica，LJE）和海带可溶性膳食纤维（dietary fiber of Laminaria japonica，LJDF），并对其成分进行分析，将以上四种活性物质组合得到海带活性物质组合物（mixture of active substances of Laminaria japonica，LJM），然后以秀丽隐杆线虫（Caenorhabditis elegans，C. elegans）为模型，探究海带活性物质对其脂质代谢相关生化指标及mRNA转录水平的影响。结果发现，LJA的成分主要包括胆碱、磷脂、嘧啶、萜类、类花生酸、类固醇、多酚和二糖；LJW主要包括分子量为368.0 kDa和1.0 kDa的两个多糖组分，其单糖组成为葡萄糖、半乳糖、岩藻糖、甘露糖和阿拉伯糖；LJE的主要成分是蛋白活性肽，包含6种蛋白（YP_006639117.1、AIW62928.1、WDS74817.1、WDS74887.1、QBF51285.1和ABB80121.1）和43种肽段；LJDF的主要成分是分子量为717.073 kDa和11.502 kDa的两个多糖组分，单糖组成为甘露糖醛酸、古罗糖醛酸、半乳糖和岩藻糖。秀丽隐杆线虫实验结果表明，海带活性物质能够显著降低线虫体内甘油三酯和丙二醛水平（P<0.01），提高总超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶水平（P<0.01）；在mRNA转录水平上，显著上调NHR-49、FAT-5、FAT-6、FAT-7、DAF-2和DAF-16基因（P<0.01），显著下调MOD-1、ACS-2和AGE-1基因（P<0.01）。海带活性物质可能是通过调节脂肪酸β氧化（NHR-49、MOD-1和ACS-2）、脂肪酸合成（FAT-5、FAT-6和FAT-7）及胰岛素（DAF-2、AGE-1和DAF-16）信号通路改善线虫脂质过氧化及降低线虫体内脂质累积。

Abstract: This study aimed to extract the active substance from Laminaria japonica (L. japonica) and evaluate their composition and hypolipidemic activity. The alcohol extract of L. japonica (LJA), water extract of L. japonica (LJW), enzymic hydrolysates of L. japonica (LJE) and dietary fiber extract of L. japonica (LJDF) were prepared by using different polar solvents combined with ultrasound-assisted alcohol extraction, water extraction, complex proteolytic enzymes and sodium carbonate acid, and their composition was analyzed. The above four active substances were mixed to obtain the total active substances of L. japonica (LJM). This study adopted C. elegans as a model organism to investigate the effects of bioactive compounds derived from L. japonica on lipid metabolism-related biochemical parameters and Messenger RNA (mRNA) transcription levels. The result revealed that the main components of LJA included choline, phospholipids, pyrimidines, terpenoids, arachidonic acid, steroids, polyphenols and disaccharides; LJW mainly consisted of two polysaccharides with molecular weights of 368.0 kDa and 1.0 kDa, with monosaccharide composition of glucose, galactose, fucose, mannose and arabinose; LJE mainly consisted of protein active peptides, containing six proteins (YP_006639117.1, AIW62928.1, WDS74817.1, WDS74887.1, QBF51285.1and ABB80121.1), 43 peptides; LJDF mainly consisted of two polysaccharides with molecular weights of 717.073 kDa and 11.502 kDa, with monosaccharide composition of mannose aldehyde, gulonose aldehyde, galactose and fucose. The result of C. elegans experiments demonstrated that the L. japonica active substances significantly reduced triglyceride (TG) and malondialdehyde (MDA) levels (P<0.01). Meanwhile, these substances also increased total superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) levels (P<0.01). At the mRNA transcription level, the L. japonica active substances significantly upregulated genes NHR-49, FAT-5, FAT-6, FAT-7, DAF-2, and DAF-16 (P<0.01). Conversely, the substances significantly downregulated MOD-1, ACS-2, and AGE-1 (P<0.01). This suggests that the L. japonica active substances improved C. elegans lipid peroxidation and reducing lipid accumulation in C. elegans through regulating fatty acid β-oxidation (NHR-49, MOD-1 and ACS-2), fatty acid synthesis (FAT-5, FAT-6 and FAT-7) and insulin (DAF-2, AGE-1, DAF-16) signaling pathways.

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