陈荫,许诚伟,童程杰,朱欣怡,谌露,李西会,牛春宇,谌燕

• 1:浙江海洋大学食品与药学学院
• 2:江西建设职业技术学院

摘要(Abstract)：

【目的】对比不同提取方法对冈村蕨藻（Caulerpa okamurae）多糖（COP）生物活性的影响，筛选高效制备工艺，揭示其抗氧化-降脂功能的协同作用机制。【方法】以冈村蕨藻为原料，采用热水浸提、超声辅助、酸提、碱提四种方法制备COP，以1,1-二苯基-2-三硝基苯肼（DPPH）和2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）自由基清除率及铁离子还原能力为指标筛选最佳提取工艺，通过响应面法优化关键参数。通过体外胆固醇吸附、胆酸盐结合实验评估降脂活性，并构建游离氨基酸（FFA）诱导的HepG2细胞非酒精性脂肪肝模型，检测细胞内过氧化氢酶（CAT）活性、超氧化物歧化酶（SOD）活性、谷胱甘肽（GSH）含量等抗氧化指标，以及总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白（LDL-C）含量等脂代谢指标。【结果与结论】4种提取方法中，热水浸提法提取的COP(COPH)质量分数（59.7%）和得率（5.2%）最高。COP-H在DPPH和ABTS自由基清除实验及铁离子还原能力实验中抗氧化效果较佳。单因素实验表明，在提取温度为80℃、提取时间为3 h、料液比[质量（g）体积（mL）比] 1∶40条件下，COP得率最高。响应面优化后的COP提取工艺为料液比为1∶39、提取温度88℃、提取时间3 h。0.125、0.250、0.500、1.000、2.000 mg/mL的COP对胆固醇及胆酸盐的结合能力均有显著的剂量依赖性（P <0.05），表明COP有良好的化学水平降脂能力。COP对HepG2细胞的安全质量浓度为25～800μg/mL。在NAFLD模型中，在安全浓度范围的COP均可正向调控细胞内CAT、SOD、GSH水平，下调细胞内ROS(P <0.05)，减轻细胞的氧化损伤；显著降低FFA处理的HepG2细胞中TC、TG以及LDL-C的含量（P <0.05），再次证明COP的抗氧化和降脂作用。研究结果可为开发海洋绿藻源功能性食品添加剂提供理论依据。

关键词(KeyWords)： 冈村蕨藻;多糖;热水提取;抗氧化;降脂

Abstract:

Keywords:

基金项目(Foundation): 浙江省自然科学基金（LTGS23D060001,LGN20C200003）;; 浙江海洋大学人才引进科研基金（JX6311130823）

作者(Author): 陈荫,许诚伟,童程杰,朱欣怡,谌露,李西会,牛春宇,谌燕

参考文献(References)：

• [1] ZHU Y L, WU S T, GUO F, et al. Structural characteristics of sulfated xylogalactomannan isolated from Caulerpa okamurae and its anticoagulant activity[J]. International Journal of Biological Macromolecules, 2024, 275:133743.
• [2]陈荫,高畅畅,董喆,等.长茎葡萄蕨藻多糖的理化性质及其对免疫抑制小鼠的调节作用[J].广东海洋大学学报, 2023, 43(4):1-9.
• [3] MANANDHAR B, KIM H J, RHYU D Y. Caulerpa okamurae extract attenuates inflammatory interaction,regulates glucose metabolism and increases insulin sensitivity in 3T3-L1 adipocytes and RAW 264.7 macrophages[J]. Journal of Integrative Medicine, 2020, 18(3):253-264.
• [4] SHARMA B R, KIM H J, KIM M S, et al. Caulerpa okamurae extract inhibits adipogenesis in 3T3-L1 adipocytes and prevents high-fat diet?induced obesity in C57BL/6 mice[J]. Nutrition Research, 2017, 47:44-52.
• [5]董喆.海葡萄(Caulerpa lentillifera)多糖的结构和生物活性的研究[D].舟山:浙江海洋大学, 2020.
• [6] HE L, YAN X T, LIANG J, et al. Comparison of different extraction methods for polysaccharides from Dendrobium officinale stem[J]. Carbohydrate Polymers, 2018,198:101-108.
• [7] XUE H K, WANG W L, BIAN J Y, et al. Recent advances in medicinal and edible homologous polysaccharides:Extraction, purification, structure, modification,and biological activities[J]. International Journal of Biological Macromolecules, 2022, 222:1110-1126.
• [8] WANG B Q, WANG Y, ZHANG J, et al. ROS-induced lipid peroxidation modulates cell death outcome:mechanisms behind apoptosis, autophagy, and ferroptosis[J].Archives of Toxicology, 2023, 97(6):1439-1451.
• [9] MUNIZ-SANTOS R, LUCIERI-COSTA G, DE ALMEIDA M A P, et al. Lipid oxidation dysregulation:an emerging player in the pathophysiology of sepsis[J].Frontiers in Immunology, 2023, 14:1224335.
• [10] LI Y X, SHENG Y, LU X C, et al. Isolation and purification of acidic polysaccharides from Agaricus blazei Murill and evaluation of their lipid-lowering mechanism[J]. International Journal of Biological Macromolecules,2020, 157:276-287.
• [11]?ZASLAN M S, SA?LAMTA?R, DEMIR Y, et al.Isolation of some phenolic compounds from Plantago subulata L. and determination of their antidiabetic, anticholinesterase, antiepileptic and antioxidant activity[J].Chemistry&Biodiversity, 2022, 19(8):e202200280.
• [12] MASEK A, CHRZESCIJANSKA E, LATOS M, et al.Influence of hydroxyl substitution on flavanone antioxidants properties[J]. Food Chemistry, 2017, 215:501-507.
• [13] CHENG H, HUANG G L. The antioxidant activities of carboxymethylated garlic polysaccharide and its derivatives[J]. International Journal of Biological Macromolecules, 2019, 140:1054-1063.
• [14]陈怡.‘红实2号’猕猴桃多糖提取优化及后熟过程中结构特征、体外生物活性变化规律的研究[D].雅安:四川农业大学, 2023.
• [15]王文帅.灵芝菌丝体多糖的结构表征及其改善血脂紊乱的相关机制探究[D].泰安:山东农业大学,2023.
• [16] LI Y C, XU B, REN X Y, et al. Inhibition of CISD2promotes ferroptosis through ferritinophagy-mediated ferritin turnover and regulation of p62–Keap1–NRF2pathway[J]. Cellular&Molecular Biology Letters, 2022,27(1):81.
• [17] HEFFERNAN S, HARNEDY-ROTHWELL P A, GITE S, et al. Blue whiting protein hydrolysates exhibit antioxidant and immunomodulatory activities in stimulated murine RAW264.7 cells[J]. Applied Sciences, 2021, 11(20):9762.
• [18] KIDGELL J T, MAGNUSSON M, DE NYS R, et al.Ulvan:a systematic review of extraction, composition and function[J]. Algal Research, 2019, 39:101422.
• [19] YANG Y Q, ZHAO M M, LIN L Z. Effects of extraction methods on structural characteristics and bile acidbinding capacities of Moringa oleifera leaf polysaccharide fractions[J]. International Journal of Food Science&Technology, 2020, 55(4):1539-1546.
• [20]廖敏,谌素华,王维民,等.张氏马尾藻多糖的体外降胆固醇活性[J].广东海洋大学学报, 2017, 37(3):80-85.
• [21] SHEEN A, AGARWAL Y, CHEAH K M, et al. Tumorlocalized catalases can fail to alter tumor growth and transcriptional profiles in subcutaneous syngeneic mouse tumor models[J]. Redox Biology, 2023, 64:102766.
• [22] HWANG I, UDDIN M J, PAK E S, et al. The impaired redox balance in peroxisomes of catalase knockout mice accelerates nonalcoholic fatty liver disease through endoplasmic reticulum stress[J]. Free Radical Biology and Medicine, 2020, 148:22-32.
• [23] TERZIEV D, TERZIEVA D, ANDONOV V, et al. T200Evaluation of superoxide dismutase in serum in patients with liver cirrhosis[J]. Clinica Chimica Acta, 2022, 530:S154-S155.
• [24] FRAMPTON G, REDDY P, JEFFERSON B, et al. Inhibition of thrombospondin-1 reduces glutathione activity and worsens acute liver injury during acetaminophen hepatotoxicity in mice[J]. Toxicology and Applied Pharmacology, 2020, 409:115323.
• [25] CAO X, LIANG Y J, CHEN H N, et al. The Chinese herbal kangxianyiai formula exerts anti-hepatocellular carcinoma effect via regulating UGDH, AKR1B10, and SLC7A11[J]. Hepatology, 2022, 76:S1329-S1330.
• [26]金燕琳.刺梨多糖的抗肿瘤活性及其羧甲基化衍生物的抗炎活性的研究[D].广州:华南理工大学,2023.
• [27]卢晓凤,骆隽言,韦明涵,等.罗汉果甜苷与黄芪多糖联用对小鼠骨骼肌抗氧化能力的影响[J].南方农业学报, 2024, 55(7):1925-1934.
• [28] LIN W L, HSIAO Y W, LIU S H, et al. Effects of lowdensity lipoprotein cholesterol on sleep apnea:Insights from a rat model of cardiovascular autonomic dysregulation[J]. Sleep Medicine, 2024, 115:76-82.