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柴胡提取物可改善杂交石斑鱼(棕点石斑鱼♂×青石斑鱼♀)的肝脂积累和免疫反应。

Dietary Radix Bupleuri extracts improves hepatic lipid accumulation and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

机构信息

Institute of Modern Aquaculture Science and Engineering, Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, Guangzhou, 510631, People's Republic of China.

Marine Fisheries Development Centre of Guangdong Province, Huizhou, 510610, People's Republic of China.

出版信息

Fish Shellfish Immunol. 2019 May;88:496-507. doi: 10.1016/j.fsi.2019.02.052. Epub 2019 Feb 28.

Abstract

In this study, two experiments were performed to explore the effect of Radix Bupleuri extracts (RBE) on growth, lipid deposition and metabolism and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) using in vitro and in vivo models. In vitro, we used 2 ml/L 20% lipid emulsion (LE)-induced steatosis in hybrid grouper primary hepatocytes, then RBE (200, 400 and 800 μg/ml) was added to the hepatocytes after (post-treatment) the incubation with 20% LE (2 ml/L) in the culture medium. We found that RBE markedly increased cell viability, which were consistent with hepatocytes morphological structure examination and lipid metabolism and immune related genes study. The above result suggested that RBE has a protective effect on this model of hepatocytes damage. In vivo, five graded levels of RBE at 0, 200, 400, 800 and 1600 mg/kg diet were supplemented to a basal diet with 15% lipid levels (high lipid), and fed to a total of 300 hybrid grouper with an average initial weight of 25.58 ± 0.05 g for 8 weeks. Growth performance, liver histology, plasma biochemical parameters, and expression of genes involved in lipid metabolism and immune-related were measured. The study indicated that dietary RBE significantly improved growth performance and feed utilization and reduced hepatosomatic index. Dietary supplementation with 200-800 mg/kg RBE diets effectively decreased serum ALP, ALT, AST and LDH contents in fish. Furthermore, adipogenesis relative mRNA levels of DGAT2, G6PD, ME1 and DGKα in fish fed 200-400 mg/kg RBE diets were lower (P < 0.05) than in those fed RBE0 diets, while dietary supplementation with 200-800 mg/kg RBE diets up-regulated lipolysis-related genes (CPT1, LPL and PPARα) expression in the liver of hybrid grouper. Moreover, dietary RBE down-regulated the expression of apoptosis-related genes (caspase-9), up-regulated the expression of antioxidant genes (CAT) and immune-related genes (MHC2, IKKα and TGF-β1). Thus, our data suggest that RBE suppressed lipid accumulation and enhanced immune capability in hybrid grouper both in vitro and in vivo. These results offer new insight into RBE as a hepatoprotective in fish.

摘要

在这项研究中,我们使用体外和体内模型来探索柴胡提取物(RBE)对杂交石斑鱼(Epinephelus lanceolatus♂×Epinephelus fuscoguttatus♀)生长、脂肪沉积和代谢以及免疫反应的影响。在体外,我们使用 2ml/L20%脂肪乳剂(LE)诱导杂交石斑鱼原代肝细胞脂肪变性,然后在培养基中用 20%LE(2ml/L)孵育后(后处理)向肝细胞中加入 RBE(200、400 和 800μg/ml)。我们发现 RBE 显著增加了细胞活力,这与肝细胞形态结构检查以及脂质代谢和免疫相关基因研究一致。上述结果表明,RBE 对这种肝细胞损伤模型具有保护作用。在体内,将 0、200、400、800 和 1600mg/kg 饮食水平的 RBE 添加到基础饮食(含 15%脂肪)中,并用含有 15%脂肪水平(高脂肪)的饮食喂养总共 300 条平均初始体重为 25.58±0.05g 的杂交石斑鱼 8 周。测量生长性能、肝脏组织学、血浆生化参数以及参与脂质代谢和免疫相关的基因表达。研究表明,饮食 RBE 可显著提高生长性能和饲料利用率,并降低肝体比。饮食补充 200-800mg/kg RBE 饮食可有效降低鱼类血清碱性磷酸酶、丙氨酸转氨酶、天冬氨酸转氨酶和乳酸脱氢酶含量。此外,在 200-400mg/kg RBE 饮食喂养的鱼中,脂肪生成相关基因(DGAT2、G6PD、ME1 和 DGKα)的相对 mRNA 水平较低(P<0.05),而饮食补充 200-800mg/kg RBE 饮食可上调肝脏中脂解相关基因(CPT1、LPL 和 PPARα)的表达。此外,饮食 RBE 下调凋亡相关基因(caspase-9)的表达,上调抗氧化基因(CAT)和免疫相关基因(MHC2、IKKα 和 TGF-β1)的表达。因此,我们的数据表明,RBE 在体外和体内均抑制了杂交石斑鱼的脂肪积累并增强了其免疫能力。这些结果为 RBE 作为鱼类的保肝剂提供了新的见解。

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