罗君,付伟杰,杨二军,黄建盛,谢瑞涛,陈刚

• 1:广东海洋大学水产学院
• 2:南方海洋科学与工程广东省实验室(湛江)
• 3:广东恒兴饲料实业股份有限公司

摘要(Abstract)：

【目的】探明饲料中添加槲皮素（quercetin,QE）对杂交石斑鱼（Epinephelus fuscoguttatus♀×Epinephelus polyphekadion♂）生长性能、肝肠抗氧化能力、肠道核因子-kappa B(Nuclear factor kappa B,NF-κB)通路相关基因表达及肠道菌群的影响。【方法】QE分别按照0(FM组)、0.80(QEL组)、1.60(QEM组)、3.20(QEH组)mmol/kg加入基础饲料，投喂体质量为（10.10±0.02）g的杂交石斑鱼幼鱼56 d，测定石斑鱼生长性能、肝肠抗氧化酶活性，用实时定量PCR分析肠道NF-κB通路相关基因表达量，用高通量测序技术分析肠道菌群结构。【结果】QEH组杂交石斑鱼增重率（WGR）、特定增长率（SGR）和终末体质量（FW）显著提高，存活率（SR）显著降低（P<0.05）。与FM组相比，QEM组肝脏超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GPX）活性显著提高（P<0.05）,QEH组肝脏GPX和过氧化氢酶（CAT）活性显著提高（P<0.05）。QE激活肠道NF-κB信号通路，上调肿瘤坏死因子（TNF-α）、TNF受体相关因子2(TRAF-2)、TNF受体超家族成员1A(TNFRSF1A)、干扰素-γ(IFN-γ)和核因子κBp65亚单位（NF-κBp65）表达水平，下调NF-κB抑制剂-α(IκB-α)的表达。与FM组相比，QEM和QEH组的运算分类单位（OTU）数目显著下降（P<0.05）,ACE、Chao1、Simpson、Shannon指数无显著差异（P>0.05）。偏最小二乘判别分析显示，FM、QEL和QEH三组间β多样性指数存在显著差异（P<0.05）。QEM组Bifidobacterium显著增加（P<0.05）。【结论】饲料中添加0.8～3.2 mmol/kg QE可降低杂交石斑鱼的存活率，增加肝脏抗氧化能力；QE可激活肠道中NF-κB信号通路，降低肠道微生物多样性，QE有肠道促炎活性。

关键词(KeyWords)： 槲皮素;杂交石斑鱼;褐点石斑鱼;清水石斑鱼;生长性能;抗氧化;NF-κB;肠道菌群

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划项目(2020YFD0900200);; 广东省科技计划项目（2016B020201009）

作者(Author): 罗君,付伟杰,杨二军,黄建盛,谢瑞涛,陈刚

参考文献(References)：

• [1] AMENYOGBE E, CHEN G, WANG Z L. Identification,characterization, and expressions profile analysis of growth hormone receptors(GHR1 and GHR2)in Hybrid grouper(Epinephelus fuscoguttatus♀×Epinephelus polyphekadion♂)[J]. Genomics, 2020, 112(1):1-9.
• [2]刘泓宇,李立贤, AYIKU Stephen,等.酵母培养物对珍珠龙胆石斑鱼生长性能、肠道形态、免疫功能和抗病力的影响[J].广东海洋大学学报, 2021, 41(3):1-11.
• [3]胡斌,宋理平,冒树泉,等.饲料中添加4种中草药制剂对大口黑鲈幼鱼生长和抗氧化能力的影响[J].广东海洋大学学报, 2019, 39(4):101-107.
• [4] LI M Y, ZHU X M, TIAN J X, et al. Dietary flavonoids from Allium mongolicum Regel promotes growth, improves immune, antioxidant status, immune-related signaling molecules and disease resistance in juvenile northern snakehead fish(Channa argus)[J]. Aquaculture, 2019, 501:473-481.
• [5] PORRAS D, NISTAL E, MARTíNEZ-FLóREZ S, et al.Protective effect of quercetin on high-fat diet-induced nonalcoholic fatty liver disease in mice is mediated by modulating intestinal microbiota imbalance and related gut-liver axis activation[J]. Free Radical Biology and Medicine, 2017,102:188-202.
• [6] DO?ATE C, BALASCH J C, CALLOL A, et al. The effects of immunostimulation through dietary manipulation in the rainbow trout; evaluation of mucosal immunity[J]. Marine Biotechnology, 2010, 12(1):88-99.
• [7] FAGGIO C, SUREDA A, MORABITO S, et al. Flavonoids and platelet aggregation:a brief review[J]. European Journal of Pharmacology, 2017, 807:91-101.
• [8] WANG W Y, SUN C X, MAO L K, et al. The biological activities, chemical stability, metabolism and delivery systems of quercetin:a review[J]. Trends in Food Science&Technology, 2016, 56:21-38.
• [9] AWAD E, AUSTIN D, LYNDON A R. Effect of black cumin seed oil(Nigella sativa)and nettle extract(Quercetin)on enhancement of immunity in rainbow trout, Oncorhynchus mykiss(Walbaum)[J]. Aquaculture, 2013, 388/389/390/391:193-197.
• [10] ZHANG J L, LIU M, CUI W, et al. Quercetin affects shoaling and anxiety behaviors in zebrafish:involvement of neuroinflammation and neuron apoptosis[J]. Fish&Shellfish Immunology, 2020, 105:359-368.
• [11] PêS T S, SACCOL E M H, LONDEROéP, et al. Protective effect of quercetin against oxidative stress induced by oxytetracycline in muscle of silver catfish[J]. Aquaculture,2018, 484:120-125.
• [12] AWAD E, AWAAD A S, ESTEBAN M A. Effects of dihydroquercetin obtained from deodar(Cedrus deodara)on immune status of gilthead seabream(Sparus aurata L.)[J]. Fish&Shellfish Immunology, 2015, 43(1):43-50.
• [13] SHIN H S, YOO J H, MIN T S, et al. Effect of quercetin on the activity and mRNA expression of antioxidant enzymes and physiological responses in olive flounder(Paralichthys olivaceus)exposed to cadmium[J]. AsianAustralasian Journal of Animal Sciences, 2010, 23(6):742-749.
• [14] XU N, FU J, WANG H, et al. Quercetin counteracts the pro-viral effect of heat shock response in grass carp cells with its therapeutic potential against Aquareovirus[J].Aquaculture Research, 2021, 52(7):3164-3173.
• [15] HARWOOD M, DANIELEWSKA-NIKIEL B, BORZELLECA J F, et al. A critical review of the data related to the safety of quercetin and lack of evidence of in vivo toxicity,including lack of genotoxic/carcinogenic properties[J].Food and Chemical Toxicology, 2007, 45(11):2179-2205.
• [16] YE G L, DONG X H, YANG Q H, et al. Low-gossypol cottonseed protein concentrate used as a replacement of fish meal for juvenile hybrid grouper(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂):effects on growth performance, immune responses and intestinal microbiota[J]. Aquaculture, 2020, 524:735309.
• [17] XIE R T, AMENYOGBE E, CHEN G, et al. Effects of feed fat level on growth performance, body composition and serum biochemical indices of hybrid grouper(Epinephelus fuscoguttatus×Epinephelus polyphekadion)[J]. Aquaculture, 2021, 530:735813.
• [18] XU Z, LI X Q, YANG H, et al. Dietary quercetin improved the growth, antioxidation, and flesh quality of grass carp(Ctenopharyngodon idella)[J]. Journal of the World Aquaculture Society, 2019, 50(6):1182-1195.
• [19] ZHAI S W, LIU S L. Effects of dietary quercetin on growth performance, serum lipids level and body composition of tilapia(Oreochromis niloticus)[J]. Italian Journal of Animal Science, 2013, 12(4):e85.
• [20] AWAD E, AWAAD A. Role of medicinal plants on growth performance and immune status in fish[J]. Fish&Shellfish Immunology, 2017, 67:40-54.
• [21] SHIN H S, YOO J H, MIN T S, et al. The effects of quercetin on physiological characteristics and oxidative stress resistance in olive flounder, Paralichthys olivaceus[J].Asian-Australasian Journal of Animal Sciences, 2010, 23(5):588-597.
• [22] SPENCER J P E, ABD EL MOHSEN M M, MINIHANE A M, et al. Biomarkers of the intake of dietary polyphenols:strengths, limitations and application in nutrition research[J]. The British Journal of Nutrition, 2008, 99(1):12-22.
• [23] PA R K K H, O C A G A R M, B O N E L L O P, e t a l.Determination of quercetin concentrations in fish tissues after feeding quercetin-containing diets[J]. Aquaculture International, 2008, 17(6):537-544.
• [24] ADER P, WESSMANN A, WOLFFRAM S. Bioavailability and metabolism of the flavonol quercetin in the pig[J].Free Radical Biology and Medicine, 2000, 28(7):1056-1067.
• [25] PRINCE P S M, SATHYA B. Pretreatment with quercetin ameliorates lipids, lipoproteins and marker enzymes of lipid metabolism in isoproterenol treated cardiotoxic male Wistar rats[J]. European Journal of Pharmacology, 2010,635(1/2/3):142-148.
• [26] DIPLOCK A T, CHARLEUX J L, CROZIER-WILLI G,et al. Functional food science and defence against reactive oxidative species[J]. The British Journal of Nutrition,1998, 80(Suppl 1):S77-S112.
• [27] ALUANI D, TZANKOVA V, KONDEVA-BURDINA M,et al.Еvaluation of biocompatibility and antioxidant efficiency of chitosan-alginate nanoparticles loaded with quercetin[J]. International Journal of Biological Macromolecules, 2017, 103:771-782.
• [28] WANG J H, ZHANG C N, ZHANG J L, et al. The effects of quercetin on immunity, antioxidant indices, and disease resistance in zebrafish(Danio rerio)[J]. Fish Physiology and Biochemistry, 2020, 46(2):759-770.
• [29] PêS T S, SACCOL E M H, OURIQUE G M, et al.Quercetin in the diet of silver catfish:effects on antioxidant status, blood parameters and pituitary hormone expression[J]. Aquaculture, 2016, 458:100-106.
• [30] KAINDL U, EYBERG I, ROHR-UDILOVA N, et al. The dietary antioxidants resveratrol and quercetin protect cells from exogenous pro-oxidative damage[J]. Food and Chemical Toxicology, 2008, 46(4):1320-1326.
• [31] RIETJENS I M C M, BOERSMA M G, VAN DER WOUDE H, et al. Flavonoids and alkenylbenzenes:mechanisms of mutagenic action and carcinogenic risk[J].M u ta tio n R e s e a rc h/F u n d a m e n ta l a n d M o le c u la r Mechanisms of Mutagenesis, 2005, 574(1/2):124-138.
• [32] CLAUS S P, GUILLOU H, ELLERO-SIMATOS S. The gut microbiota:a major player in the toxicity of environmental pollutants?[J]. Npj Biofilms and Microbiomes, 2016, 2:16003.
• [33] ZHANG J M, RHO Y, KIM M Y, et al. TAK1 in the AP-1pathway is a critical target of Saururus chinensis(Lour.)Baill in its anti-inflammatory action[J]. Journal of Ethnopharmacology, 2021, 279:114400.
• [34] G A B R I E L E M, P U C C I L,áRVAY J, e t a l. A n t iinflammatory and antioxidant effect of fermented whole wheat on TNFα-stimulated HT-29 and NF-κB signaling pathway activation[J]. Journal of Functional Foods, 2018,45:392-400.
• [35] MEYER M, SCHRECK R, BAEUERLE P A. H2O2and antioxidants have opposite effects on activation of NF-kappa B and AP-1 in intact cells:AP-1 as secondary antioxidant-responsive factor[J]. The EMBO Journal, 1993,12(5):2005-2015.
• [36] YANG J X, MARIA T C, ZHOU B, et al. Quercetin improves immune function in Arbor Acre broilers through activation of NF-κB signaling pathway[J]. Poultry Science, 2020, 99(2):906-913.
• [37] GIRI S S, SUKUMARAN V, PARK S C. Effects of bioactive substance from turmeric on growth, skin mucosal immunity and antioxidant factors in common carp, Cyprinus carpio[J]. Fish&Shellfish Immunology, 2019, 92:612-620.
• [38] NOOTASH S, SHEIKHZADEH N, BARADARAN B, et al.Green tea(Camellia sinensis)administration induces expression of immune relevant genes and biochemical parameters in rainbow trout(Oncorhynchus mykiss)[J].Fish&Shellfish Immunology, 2013, 35(6):1916-1923.
• [39] DELZENNE N M, CANI P D. Interaction between obesity and the gut microbiota:relevance in nutrition[J]. Annual Review of Nutrition, 2011, 31:15-31.
• [40] BRAUNE A, BLAUT M. Bacterial species involved in the conversion of dietary flavonoids in the human gut[J]. Gut Microbes, 2016, 7(3):216-234.
• [41] HE Y F, GUO X W, TAN B P, et al. Replacing fish meal with fermented rice protein in diets for hybrid groupers(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂):effects on growth, digestive and absorption capacities,inflammatory-related gene expression, and intestinal microbiota[J]. Aquaculture Reports, 2021, 19:100603.
• [42] LIU Y, CHEN Z C, DAI J H, et al. The protective role of glutamine on enteropathy induced by high dose of soybean meal in turbot, Scophthalmus maximus L[J]. Aquaculture,2018, 497:510-519.
• [43] RIMOLDI S, TEROVA G, ASCIONE C, et al. Next generation sequencing for gut microbiome characterization in rainbow trout(Oncorhynchus mykiss)fed animal by-product meals as an alternative to fishmeal protein sources[J]. PLoS One, 2018, 13(3):e0193652.
• [44] JONES S E, LENNON J T. Dormancy contributes to the maintenance of microbial diversity[J]. PNAS, 2010, 107(13):5881-5886.
• [45] XIONG J B, WANG K, WU J F, et al. Changes in intestinal bacterial communities are closely associated with shrimp disease severity[J]. Applied Microbiology and Biotechnology,2015, 99(16):6911-6919.
• [46] TILG H, ADOLPH T E, GERNER R R, et al. The intestinal microbiota in colorectal cancer[J]. Cancer Cell, 2018, 33(6):954-964.
• [47] JIANG F, LIU M H, WANG H D, et al. Wu Mei Wan attenuates CAC by regulating gut microbiota and the NF-κB/IL6-STAT3 signaling pathway[J]. Biomedicine&Pharmacotherapy, 2020, 125:109982.
• [48]郑有坤,刘凯,熊子君,等.药用植物内生放线菌多样性及天然活性物质研究进展[J].中草药, 2014, 45(14):2089-2099.
• [49]谭蓉,陈郑珊,杨栋,等.肠道菌群介导高蛋白饮食引起小鼠肠道炎症研究[J].解放军预防医学杂志, 2020, 38(10):81-83.
• [50] SHIN N R, WHON T W, BAE J W. Proteobacteria:microbial signature of dysbiosis in gut microbiota[J].Trends in Biotechnology, 2015, 33(9):496-503.
• [51] YANG B, CHEN H Q, GAO H, et al. Bifidobacterium breve CCFM683 could ameliorate DSS-induced colitis in mice primarily via conjugated linoleic acid production and gut microbiota modulation[J]. Journal of Functional Foods,2018, 49:61-72.
• [52]孙杰,王丽红,官娜,等.灭活的双歧杆菌对小鼠肠液中分泌型IgA水平的影响[J].大连大学学报, 2012, 33(3):53-55.
• [53]张亮,王世达,谢方,等.肠易激综合征大鼠肠道菌群和氨基酸代谢的变化[J].营养学报, 2018, 40(3):240-244.
• [54] COLLADO M C, ISOLAURI E, LAITINEN K, et al. Effect of mother's weight on infant's microbiota acquisition,composition, and activity during early infancy:a prospective follow-up study initiated in early pregnancy[J]. The American Journal of Clinical Nutrition, 2010, 92(5):1023-1030.