高脂饲料添加柚皮苷对大口黑鲈肝脏脂质代谢和内质网应激的影响

Effects of dietary naringin supplementation in high fat on hepatic lipid metabolism and endoplasmic reticulum stress in largemouth bass (Micropterus salmoides)

  • 摘要:
    目的 探究高脂和柚皮苷(Nar)对大口黑鲈脂质代谢和内质网应激的影响,为植物提取物在水产饲料中的应用提供科学依据。
    方法 以大口黑鲈幼鱼为研究对象,设置对照组(10.5%粗脂肪)、高脂组(18.5%粗脂肪)、对照组+0.125%Nar、高脂组+0.125%Nar、对照组+0.25%Nar和高脂组+0.25%Nar 6个饲料组,每组3个重复,实验持续10周。通过苏木精-伊红和油红O染色观察肝脏组织学和脂肪滴的变化;甘油三酯(TG)分析试剂盒检测肝脏中的TG含量;荧光定量PCR和蛋白质免疫印迹测定相关基因和蛋白表达水平。
    结果 在高脂组中,肝脏组织受到损伤产生空泡化现象,通过油红O染色,结果显示,肝脏中有大量脂滴,肝脏TG含量和脂质合成酶相关基因(scd1、fasacca)和转录因子SREBP1、PPARγ的mRNA表达水平升高。脂质分解酶相关基因(cpt1、cpt2、atglhsl)和转录因子PPARα的mRNA表达水平降低。添加柚皮苷可降低肝脏中TG含量、脂质合成相关基因mRNA的表达水平,增加脂质分解相关基因mRNA的表达水平。与对照组相比,高脂组中肝脏透射电镜显示,内质网发生明显肿胀,内质网应激激活的相关通路基因(ire1、xbp1、elf2αatf4、atf6和grp78)的mRNA水平和内质网应激标志蛋白GRP78表达量升高。添加Nar可缓解肝脏内质网肿胀,同时内质网应激激活的相关通路基因的mRNA水平和内质网应激标志蛋白GRP78表达量下降。
    结论 添加柚皮苷可以缓解高脂导致的大口黑鲈的肝脏脂质沉积和内质网应激。本研究为水产养殖过程中高脂饲料饲喂提供了一定的理论基础,并为植物提取物在水产饲料中的添加提供参考。

     

    Abstract: This study investigated the effects of high-fat diet (HFD) and naringin(Nar) supplementation on hepatic lipid metabolism and endoplasmic reticulum stress in largemouth bass (Micropterus salmoides). The aim was to provide a scientific basis for the application of plant extracts as aquaculture feed additives. The experiment used juvenile M. salmoides as the research subjects and set up six groups of feed, namely the control group (10.5% crude lipid), the high-fat group (18.5% crude lipid), the control group + 0.125% Nar, the high-fat group + 0.125% Nar, the control group + 0.25% Nar, and the high-fat group + 0.25% Nar. Each group had three replicates, and the experiment lasted for ten weeks. The results showed that in the HFD group, liver tissue was damaged and vacuolated, and oil O red staining revealed a large number of lipid droplets in the liver. The mRNA expression levels of lipid synthesis-related genes (scd1, fas, acca) and transcription factors SREBP1 and PPARγ were increased, while the mRNA expression levels of lipid decomposition-related genes (cpt1, cpt2, atgl, hsl) and transcription factor PPARα were decreased. The addition of naringin reduced the TG content in the liver and the mRNA expression levels of lipid synthesis-related genes, while increasing the mRNA expression levels of lipid decomposition-related genes. Additionally, compared with the control group, the transmission electron microscopy of the liver in the HFD group showed obvious swelling of the endoplasmic reticulum, and the mRNA levels of endoplasmic reticulum stress-related pathway genes (ire1, xbp1, elf2α, atf4, atf6, grp78) and the expression level of the endoplasmic reticulum stress marker protein GRP78 were increased. The addition of naringin alleviated the swelling of the endoplasmic reticulum in the liver and decreased the mRNA levels of endoplasmic reticulum stress-related pathway genes and the expression level of the endoplasmic reticulum stress marker protein GRP78. In conclusion, dietary naringin supplementation can alleviate HFD-induced hepatic lipid deposition and endoplasmic reticulum stress in M. salmoides. This provides a theoretical basis for the utilization of high-fat feed in aquaculture and offers reference into the addition of plant extracts in aquaculture feed.

     

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