This study utilized large yellow croaker bones as raw materials to prepare chondroitin sulfate through processes including dilute alkali enzymatic hydrolysis, ethanol precipitation, and dialysis for impurity removal.The structural characteristics of large yellow croaker chondroitin sulfate (LCS) were further investigated.Subsequently, a hyperlipidemic zebrafish model was established to explore its hypolipidemic activity.Through the determination of physical and chemical properties, ultraviolet spectrophotometer, Fourier transform infrared spectrometer, high performance gel permeation chromatography, and 1H nuclear magnetic resonance spectroscopy, it was found that the extraction rate and content of CS in large yellow croaker bone reached 0.46% and 92.59%, respectively, and the content of glucuronic acid and protein in LCS was 35.64% and 1.27%, respectively.The functional group composition was similar to that of CS standard.The weight-average molecular weight of LCS was 33 874 Da, and the degree of sulfation occurred at the point of C-6 and C-4 was similar.The results of oil red O staining showed that the high-fat diet successfully induced hyperlipidemia in zebrafish.After intervention with different mass concentrations of LCS (40, 80, and 200 μg/mL), the lipid level significantly decreased (P<0.05).Hematoxylin-eosin staining indicated that with the increase in LCS mass concentration, the arrangement of hepatocytes in livers gradually changed from loose to tight, and the fat vacuoles gradually decreased.LCS could significantly reduce the levels of low-density lipoprotein cholesterol, total cholesterol, and triglycerides in hyperlipidemic zebrafish (P<0.05), and increase the level of high-density lipoprotein cholesterol (P<0.05).The expression levels of genes related to lipid metabolism in zebrafish were determined using quantitative real-time polymerase chain reaction.It was found that LCS could alleviate lipid metabolism disorder in the hyperlipidemic zebrafish model by down-regulating the expression of lipid synthesis-related genes (Srebf1, Acaca, Fasn, Hmgcrb) and up-regulating the expression of lipid decomposition genes (Pparg, Rora, Ldlr, Cyp7a1a).
[1] 农业农村部渔业渔政管理局. 2024 中国渔业统计年鉴[M].北京:中国农业出版社, 2024.
Fishery Bureau of Ministry of Agriculture of the People’s Republic of China.China Fishery Statistical Yearbook[M].Beijing:China Agriculture Press, 2024.
[2] 刘梦遥. 清蒸大黄鱼加工工艺及其贮藏品质的研究[D].福州:福建农林大学, 2023.
LIU M Y.Study on the steaming processing of Larimichthys crocea products and its quality changes during storage[D].Fuzhou:Fujian Agriculture and Forestry University, 2023.
[3] 窦鑫. 大黄鱼鱼肝油的酶法制取与脱腥研究[D].上海:上海海洋大学, 2022.
DOU X.Enzymatic preparation and deodorization of large yellow croaker liver oil[D].Shanghai:Shanghai Ocean University, 2022.
[4] SHANG Q S, SHI J J, SONG G R, et al.Structural modulation of gut microbiota by chondroitin sulfate and its oligosaccharide[J].International Journal of Biological Macromolecules, 2016, 89:489-498.
[5] YUAN Q X, SHI X, MA H Q, et al.Recent progress in marine chondroitin sulfate, dermatan sulfate, and chondroitin sulfate/dermatan sulfate hybrid chains as potential functional foods and therapeutic agents[J].International Journal of Biological Macromolecules, 2024, 262:129969.
[6] MISHRA S, GANGULI M.Functions of, and replenishment strategies for, chondroitin sulfate in the human body[J].Drug Discovery Today, 2021, 26(5):1185-1199.
[7] 宋琳, 宋晓, 薛长湖, 等.海地瓜岩藻聚糖与硫酸软骨素双水相分离方法的研究[J].食品与发酵工业, 2023, 49(19):183-189.
SONG L, SONG X, XUE C H, et al.Study on the PEG-salt aqueous two-phase method for the separation of sulfated fucan and chondroitin sulfate in Acaudina molpadioides[J].Food and Fermentation Industries, 2023, 49(19):183-189.
[8] SHEN Q S, GUO Y J, WANG K Y, et al.A review of chondroitin sulfate’s preparation, properties, functions, and applications[J].Molecules, 2023, 28(20):7093.
[9] 刘良禹, 汪静汶, 朱思洁, 等.超声辅助-碱-酶法提取鲟鱼硫酸软骨素工艺及其结构分析[J].食品科技, 2023, 48(10):108-116.
LIU L Y, WANG J W, ZHU S J, et al.Ultrasonic assisted-alkaline-enzymatic extraction and structural analysis of chondroitin sulfate from sturgeon[J].Food Science and Technology, 2023, 48(10):108-116.
[10] YANG J, SHEN M Y, WU T, et al.Physicochemical, structural characterization, and antioxidant activities of chondroitin sulfate from Oreochromis niloticus bones[J].Food Science and Human Wellness, 2023, 12(4):1102-1108.
[11] 郑昕. 鸡胸软骨酶解物中硫酸软骨素含量测定及结构功能探究[D].广州:华南理工大学, 2021.
ZHENG X.Determination of chondroitin sulfate in enzymatic hydrolysate of chicken breast cartilage and research on its structure and function[D].Guangzhou:South China University of Technology, 2021.
[12] 周畅. 鳐硫酸软骨素及其降解产物的结构表征和抗氧化活性研究[D].保定:河北农业大学, 2020.
ZHOU C.Structural characterization and antioxidant activity of skate chondroitin sulfate and its degradation products[D].Baoding:Hebei Agricultural University, 2020.
[13] 任珍芸, 陈晓航, 王玺, 等.硫酸-咔唑微孔板法检测肺炎链球菌荚膜多糖中糖醛酸含量[J].微生物学免疫学进展, 2017, 45(2):36-41.
REN Z Y, CHEN X H, WANG X, et al.Determination of uronic acid content in pneumococcal polysaccharide by sulfate-carbazole method in microplate[J].Progress in Microbiology and Immunology, 2017, 45(2):36-41.
[14] 左格格. 罗非鱼加工副产物不同部位硫酸软骨素的制备、结构表征及抗衰老活性[D].湛江:广东海洋大学, 2022.
ZUO G G.Preparation, structural characterization and antiaging activity of chondroitin sulfate from different parts of tilapia by-product[D].Zhanjiang:Guangdong Ocean University, 2022.
[15] 叶琳弘. 鱿鱼软骨中硫酸软骨素的提取、纯化及降血脂活性的研究[D].福州:福建农林大学, 2015.
YE L H.Studied on extraction, purification and hypolipidemic activity of chondroitin sulfate from cartilage of squid[D].Fuzhou:Fujian Agriculture and Forestry University, 2015.
[16] ZHOU C, MI S, LI J, et al.Purification, characterisation and antioxidant activities of chondroitin sulphate extracted from Raja porosa cartilage[J].Carbohydrate Polymers, 2020, 241:116306.
[17] MUCCI A, SCHENETTI L, VOLPI N.1H and13C nuclear magnetic resonance identification and characterization of components of chondroitin sulfates of various origin[J].Carbohydrate Polymers, 2000, 41(1):37-45.
[18] ABDALLAH M M, FERNÁNDEZ N, MATIAS A A, et al.Hyaluronic acid and chondroitin sulfate from marine and terrestrial sources:Extraction and purification methods[J].Carbohydrate Polymers, 2020, 243:116441.
[19] SUDUN, LIU S L, XIAO C, et al.Probiotic strains improve high-fat diet-induced hypercholesterolemia through modulating gut microbiota in ways different from atorvastatin[J].Food & Function, 2019, 10(9):6098-6109.
[20] 杨雯. 不同剂量阿托伐他汀在动脉粥样硬化性血管内皮功能保护中的应用效果分析[J].吉林医学, 2021, 42(2):411-413.
YANG W.Effect analysis of different doses of atorvastatin in the protection of atherosclerotic vascular endothelial function[J].Jilin Medical Journal, 2021, 42(2):411-413.
[21] YU H, YI X Z, GAO X, et al.Tilapia-head chondroitin sulfate protects against nonalcoholic fatty liver disease via modulating the gut-liver axis in high-fat-diet-fed C57BL/6 mice[J].Foods, 2022, 11(7):922.
[22] 杜蕊, 李娜, 赵彩杰, 等.HMG-CoA还原酶抑制剂联合阿托伐他汀对冠心病大鼠的干预及降血脂作用[J].中国老年学杂志, 2023, 43(1):128-132.
DU R, LI N, ZHAO C J, et al.Intervention of HMG-CoA reductase inhibitor combined with atorvastatin on coronary heart disease rats and its hypolipidemic effect[J].Chinese Journal of Gerontology, 2023, 43(1):128-132.
[23] XIONG S L, LI A L, JIN Z Y, et al.Effects of oral chondroitin sulfate on lipid and antioxidant metabolisms in rats fed a high-fat diet[J].Journal of Food Biochemistry, 2007, 31(3):356-369.
[24] AREGGER M, LAWSON K A, BILLMANN M, et al.Systematic mapping of genetic interactions for de novo fatty acid synthesis identifies C12orf49 as a regulator of lipid metabolism[J].Nature Metabolism, 2020, 2(6):499-513.
[25] YANG X F, SHANG D J.The role of peroxisome proliferator-activated receptor γ in lipid metabolism and inflammation in atherosclerosis[J].Cell Biology International, 2023, 47(9):1469-1487.
[26] LEE J M, KIM H, BAEK S H.Unraveling the physiological roles of retinoic acid receptor-related orphan receptor α[J].Experimental & Molecular Medicine, 2021, 53(9):1278-1286.
[27] SONG Y O, KIM M, WOO M, et al.Chondroitin sulfate-rich extract of skate cartilage attenuates lipopolysaccharide-induced liver damage in mice[J].Marine Drugs, 2017, 15(6):178.
[28] AKSOY M O, BILINSKA A, STACHOWIAK M, et al.Deciphering the role of the SREBF1 gene in the transcriptional regulation of porcine adipogenesis using CRISPR/Cas9 editing[J].International Journal of Molecular Sciences, 2024, 25(23):12677.
[29] RUIZ R, JIDEONWO V, AHN M, et al.Sterol regulatory element-binding protein-1 (SREBP-1) is required to regulate glycogen synthesis and gluconeogenic gene expression in mouse liver[J].Journal of Biological Chemistry, 2014, 289(9):5510-5517.
[30] ZHAO Y, PENG L, YANG L C, et al.Wedelolactone regulates lipid metabolism and improves hepatic steatosis partly by AMPK activation and up-regulation of expression of PPARα/LPL and LDLR[J].PLoS One, 2015, 10(7):e0132720.
[31] SONG J P, LU X Y, LIU D H, et al.Fucogalactan sulfate (FS) from Laminaria japonica regulates lipid metabolism in diet-induced humanized dyslipidemia mice via an intestinal FXR-FGF19-CYP7A1/CYP8B1 pathway[J].Journal of Agricultural and Food Chemistry, 2023, 71(38):14027-14037.
