bassianolide and Fatty-Liver

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.

Topics: Animal Feed; Animals; Apoptosis; Bass; Caspase 9; Cell Survival; Cells, Cultured; Dietary Supplements; Fatty Liver; Female; Fish Proteins; Hepatocytes; Hybridization, Genetic; Lipid Metabolism; Lipolysis; Liver; Male; Plant Extracts; Plant Roots; RNA, Messenger

A 10-week growth trial was conducted to investigate the effects of replacing dietary fishmeal with plant proteins on nutrition metabolism, immunity, inflammation and apoptosis responses in liver tissues of Japanese seabass, Lateolabrax japonicas (initial body weight = 10.42 ± 0.01 g). Two isonitrogenous and isoenergetic diets were formulated. A basal diet containing 54% fishmeal (FM), whereas another diet was prepared by totally replacing FM with a plant protein blend (PP) composed with soybean protein concentrate and cottonseed protein concentrate. Although essential amino acids, fatty acids, and available phosphorus had been balanced according to the FM diet profile, the significantly lower growth performance, metabolic disorder, and fatty liver symptom were observed in the PP group. Compared with the FM group, fish in the PP group showed significantly lower plasma free EAA level and PPV. Glucose metabolism disorder was expressed as the uncontrollable fasting glycolysis and pyruvate aerobic oxidation at postprandial 24 h with significantly up-regulated GK, PK and PDH genes expression, which potentially over-produced acetyl-CoA as the substrate for protein and lipid synthesis. Significantly reduced plasma GLU, but increased GC level, along with very significantly reduced liver GLY storage could be observed in the PP group. Plasma TG and hepatic NEFA contents were significantly decreased, but the hepatic TC content was very significantly increased in the PP group, in addition, hepatocyte vacuolation appeared. The significantly up-regulated cholesterol synthesis gene (HMGCR) expression but down-regulated bile acid synthesis gene (CYP7A1) expression could be the main reason for the fatty liver induced by cholesterol accumulation. The reduced plasma IgM content accompanied by the up-regulated mRNA levels of pro-inflammatory cytokines (TNFα and IL1β) and activated apoptosis signals of liver tissues were found in the PP group. The hyperthyroidism (higher plasma T3 and T4) and the accelerated energy metabolism rate decreased the growth performance in the PP group. The activated p65NF-kB may promote the hepatocytes apoptosis via the extrinsic pathway (caspase8/caspase3). Simultaneously, a "self-saving" response could be observed that activated cAMP promoted the lipolysis/β-oxidation process and up-regulated gene expression of anti-inflammatory cytokine IL10 via promoting CREB expression, further inhibited the over-phosphorylation of JNK protein, which might impe

Topics: Animal Feed; Animals; Basal Metabolism; Bass; Diet; Fatty Liver; Fish Diseases; Liver; Metabolic Diseases; Plant Proteins, Dietary; Signal Transduction

Excessive lipid accumulation and chemical abuse can induce fatty liver diseases in fish, but the underlying mechanism and therapies are unknown. The present study aims to evaluate the effects of Xiaochaihu Decoction (XCHD) on the growth performance, lipid metabolism and antioxidant function of hybrid grouper in vitro and in vivo, and provide evidence as to whether it can be potentially used as a medicine for liver diseases in aquaculture. In vitro, steatosis model of hybrid grouper primary hepatocytes were incubated for 48 h in control or lipid emulsion (LE)-containing medium with or without 24 h post-treatment with XCHD. XCHD treatment reversed the LE-induced intracellular lipid accumulation, cell viability and hepatocytes morphological structure. In vivo, a total of 300 hybrid grouper with an average initial weight of 25.43 ± 0.18 g were fed diets containing five graded levels of XCHD at 150-1200 mg/kg diet for 8 weeks. After that, a challenge trial was conducted by injection of D-GalN/LPS to induce liver injury. As a result, dietary supplementation with 150-300 mg/kg XCHD diets can significant improve growth performance and feed utilization (P < 0.05). Dietary XCHD down-regulated the expression of lipogenic-related genes (G6PD, DGAT2 and ME1) and up-regulated lipolysis-related genes (ATGL, PPARα and LPL) expression in the liver of hybrid grouper. Livers challenged with D-GalN/LPS exhibited extensive areas of vacuolization with the disappearance of nuclei and the loss of hepatic architecture. These pathological alterations were ameliorated by XCHD treatment. XCHD significantly down-regulated the D-GalN/LPS induced apoptosis-related genes caspase-3, caspase-9 and p53 mRNA expression and up-regulated the antioxidant-related genes CAT and MnSOD mRNA expression in dose dependent manner, respectively. XCHD potently reduced hepatic lipid accumulation and enhanced antioxidant capability in hybrid grouper and may be a potential fish-feed additive to prevent fatty liver diseases onset and progression.

Topics: Animal Feed; Animals; Antioxidants; Apoptosis; Bass; Diet; Dietary Supplements; Drugs, Chinese Herbal; Fatty Liver; Female; Fish Diseases; Galactosamine; Lipid Metabolism; Lipopolysaccharides; Liver; Male