枳实提取物对越冬后草鱼肝肠代谢、抗氧化及免疫的影响

CITRUS AURANTIUM EXTRACT ON THE HEPATO-INTESTINAL METABOLISM, ANTIOXIDANT CAPACITY, AND IMMUNE RESPONSE OF GRASS CARP (CTENOPHARYNGODON IDELLA) AFTER OVERWINTERING

  • 摘要: 为探究高棉菜粕饲料中添加枳实提取物对越冬及越冬后复投喂草鱼的形体指标、消化酶活性、肝肠组织形态、免疫及抗氧化性能的影响, 本研究分别在越冬阶段(实验一)与越冬后恢复阶段(实验二)开展连续养殖试验。实验一设置越冬前对照组(CON)、越冬后对照组(HCON)、越冬后高棉粕饲料组(HCA0)及越冬后0.15%枳实提取物组(HCA1); 实验二设置高棉粕饲料组(FCA0)、0.15% (FCA1)及0.5% (FCA2)枳实提取物组, 恢复投喂期为2周。结果表明, 越冬胁迫显著诱导了草鱼肠道生理功能障碍, 核心表现为淀粉酶、脂肪酶及胰蛋白酶活性的显著下调, 且紧密连接基因的表达紊乱(zo-1cl-b/c表达下调, cl-12/15mlck表达上调), 并伴随肝肠组织结构的严重破坏(肠道绒毛高度下降、杯状细胞减少; 肝脏出现核偏移、肝窦扩张及空泡化)及血清生化指标的紊乱(AST、ALT升高; TG、TC降低)。同时还引发肝肠Nrf2/Keap1通路失衡(catCuZnsodgpx1grnrf2表达水平下降, keap1表达水平上升), 进而激活NF-κB信号通路调控炎症因子差异表达(tnf-αil-8上升; il-15tgf-β1下降)。在不同营养组别中, 高棉菜粕组(HCA0)机体损伤最严重, 而添加0.15%枳实提取物(HCA1)组其各项生理指标与对照组水平相当。进入复投喂期后, 机体生理功能的恢复呈现显著的枳实提取物剂量依赖性, 尤其是高剂量组(FCA2)在消化酶活性的重塑上表现出最强效力, 并同步带动了物理屏障修复、抗氧化防御提升及免疫紊乱的逆转。综上, 饲料中添加枳实提取物可通过保护并激活消化酶系统, 协同调节氧化应激与免疫反应, 从而缓解低温应激造成的肝肠损伤, 维持高棉菜粕摄食水平下越冬草鱼的生理稳态, 并能够在短期复投喂中表现出明显的保护效应。

     

    Abstract: To evaluate the effects of supplementation in high cottonseed and rapeseed meal diets on overwintering stress and post-winter refeeding in grass carp (Ctenopharyngodon idella), two consecutive feeding trials were conducted during the overwintering period (Experiment 1) and the recovery phase (Experiment 2). Experiment 1 included a pre-overwintering control (CON), a post-overwintering control (HCON), a high cottonseed and rapeseed meal diet group (HCA0), and a diet supplemented with 0.15% Citrus aurantium extract (HCA1). Experiment 2 involved refeeding for 2 weeks with a cottonseed meal diet (FCA0) or diets supplemented with 0.15% (FCA1) or 0.5% (FCA2) extract. The results demonstrated that overwintering stress significantly triggered intestinal physiological dysfunction in grass carp, primarily characterized by suppressed digestive enzyme activities (amylase, lipase, and trypsin), tight junction dysregulation (zo-1 and cl-b/c downregulation; cl-12/15 and mlck upregulation), alongside severe hepato-intestinal architectural disruption and altered serum biochemical profiles (elevated AST/ALT; diminished TG/TC). Concurrently, overwintering stress suppressed the Nrf2/Keap1 antioxidant signaling pathway (nrf2, cat, CuZnsod, gpx1, and gr downregulation; keap1 upregulation), while activating NF-κB–mediated inflammatory responses (tnf-α and il-8 upregulation; il-15 and tgf-β1 downregulation). The HCA0 group showed the most severe impairment, whereas the HCA1 group maintained physiological indices comparable to the CON group. Post-refeeding, CAE exhibited a dose-dependent recovery, with the 0.5% dose (FCA2) most effectively restoring enzymatic activity, physical barrier integrity, and antioxidant/immune status. Collectively, dietary CAE alleviates cold-induced hepato-intestinal injury by preserving digestive enzyme function and coordinately regulating oxidative stress and immune responses, thereby maintaining physiological homeostasis in overwintering grass carp fed high cottonseed and rapeseed meal diets and conferring pronounced protective effects during short-term refeeding.

     

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