鱼类microRNA研究进展

microRNA是一种大小为18~25个核苷酸的非编码RNA,调控靶基因的转录后表达,广泛参与多种生理过程。本研究简要综述了microRNA生物学特性、转录后调控机制,详细讨论了第二代测序技术的相关应用、microRNA在鱼类中的表达模式、功能研究,包括渗透压调节、生殖、发育、生长、免疫、代谢等。现有的研究表明,miR-200和miR-30具有调控血浆离子浓度和渗透压平衡的功能;Dicer1是胚胎发育所必需的,在原肠胚时期,miR-20a调节头、眼、脊柱、体节的形态发生,miR-92调控内胚层的形成,miR-10影响体轴形成,miR-219影响头部和尾部细胞的凋亡;miR-122、miR-30、miR-145分别影响肝细胞成熟、肝胆管和肠道发育,miR-138和miR-143调控心脏发育,miR-126、miR-150、miR-451调控血小板、血细胞的发育和成熟,miR-10和miR-126影响血管生成,miR-200、miR-183分别影响嗅觉、听觉系统的发育,多个microRNA参与视网膜、肌肉、鱼鳍、肝脏等器官的再生;miR-133a/b、miR-206抑制肌肉细胞分裂,miR-214、miR-499、miR-199、miR-3906调控快肌纤维和慢肌纤维的生成;miR-430促进原始生殖细胞的生成;多个microRNA参与到禁食和再投喂调控的分解代谢和合成代谢平衡;此外,microRNA通过对免疫因子的调控参与到先天免疫、低氧适应、体色调控、应激反应、肌间刺形成等生理过程中。本研究通过对这些结果进行总结和讨论分析,将有助于对鱼类microRNA研究领域现状的了解和把握,促进鱼类非编码RNA研究的深入。     

Preliminary study to evaluate the effects of dietary bile acids on growth performance and lipid metabolism of juvenile genetically improved farmed tilapia (Oreochromis niloticus) fed plant ingredient‐based diets

A 9‐week feeding experiment was conducted to investigate the effects of dietary bile acids (BAs) on juvenile genetically improved farmed tilapia (GIFT) (Oreochromis niloticus) based on the evaluations of growth performance and parameters relevant to lipid metabolism. Each of five vegetable protein‐based diets containing BAs at a level of 0, 0.05, 0.15, 0.45 or 1.35 g/kg diet was fed to three replicates with 40 fish (8.2 g per fish). The results showed that weight gain (WG) increased significantly with the increase in BAs from 0 to 0.15 g/kg diet and then decreased significantly at a higher BA supplementation. Dietary BAs significantly reduced the crude lipid content in the whole body, muscle and liver tissue of GIFT. Fish fed diet with 1.35 g BAs/kg diet developed serious nuclear migration and vacuolization in hepatocytes. Gall bladder appeared to contain white solid and has fragile capsules. Dietary BA supplementation had significant effects on serum biochemical indices and activities of lipid metabolism enzymes in liver and intestine. In conclusion, dietary bile acid supplementation (0.15 g/kg) can facilitate the lipid metabolism and therefore promote the growth of tilapia. However, overdosed dietary BAs induced gallstone development, disrupted lipid metabolism and depressed the growth performances of GIFT.     

猪油对罗非鱼生长及餐后脂质代谢的影响

在实用饲料配方的基础上分别添加0%、5%、10%猪油组成试验饲料饲喂罗非鱼(Oreochromis niloticus)(初重43 g)8周,比较研究猪油不同添加水平对罗非鱼生长及餐后脂质代谢的影响。结果表明,罗非鱼的末重(FBW)、特定生长率(SGR)、增重率(WGR)和饲料效率(FE)随着饲料中猪油水平的增加而显著升高。而摄食量(FI)表现出相反的变化趋势,即高水平猪油组显著低于对照组。随猪油添加水平增加,罗非鱼肝胰脏超氧化物歧化酶(SOD)、谷丙转氨酶(ALT)和谷草转氨酶(AST)活力显著降低,而肝脏丙二醛(MDA)含量显著增加。血清中甘油三酯(TG)、胆固醇(CHO)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)和极低密度脂蛋白胆固醇(VLDL-C)含量均随饲料中猪油水平增加而显著升高。各试验组罗非鱼血清中CHO、TG、LDL-C和VLDL-C含量随时间推移均表现出先升高后降低的变化趋势,TG和VLDL-C的峰值出现在餐后6h,而CHO和LDL-C峰值在餐后9 h。但HDL-C表现出相反的变化趋势,即先降低后升高,在餐后6 h达到最低值。总之,饲料中添加不同水平猪油会促进罗非鱼生长,但会影响罗非鱼的肝功能以及餐后脂质代谢,罗非鱼抗氧化能力也相应受到影响。     

Effect of dietary protein/carbohydrate ratio on activities of hepatic enzymes involved in the amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.)

This study investigated the influence of different dietary protein/carbohydrate ratios on activities of enzymes involved in the amino acid metabolism as well as on growth performance and body composition of Nile tilapia (Oreo- chromis niloticus). Fish were fed semi-synthetic diets differing in their protein/carbohydrate ratio. The diets were formulated to be isolipidic and isoenergetic (on the basis of metabolizable energy). Fish were reared individually. After a prefeeding phase, tilapia were randomly assigned to three groups and were fed the experimental diets at a relatively low level. The study lasted 9 weeks. Proximate composition, gross energy content, and activities of aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) in the liver of fish were determined. While the mean growth rate and feed conversion efficiency did not differ significantly among the feeding groups, the body composition showed significant differences. The dietary protein/carbohydrate ratio had no significant effect on the protein and energy gain, but the lipid gain was significantly higher in tilapia fed the low-protein diet. The activities of ASAT and ALAT per gram of liver and per gram of soluble protein were significantly increased in fish fed the high-protein diet. There was a significant positive correlation between the activities of ASAT and ALAT. The activities of both enzymes in the liver were significantly higher in fish with a higher protein gain. Measuring the activities of enzymes involved in the amino acid metabolism can provide more information about the metabolic utilized dietary protein.     

The effect of hepatic passage on postprandial plasma lipid profile of rainbow trout (Oncorhynchus mykiss) after a single meal

For the first time, pre‐ and post‐hepatic plasma lipid profiles were monitored following a single meal in a free‐swimming, non‐anaesthetized fish. Rainbow trout (Oncorhynchus mykiss; 700–1500 g; 10 °C) were equipped with cannulae in the dorsal aorta (DA) and hepatic portal vein (HPV). Simultaneous blood samples, taken from both cannulae at 0, 3, 6, 12, 24 and 48 h postprandial, revealed the time course of the plasma lipid profiles following a single meal (1% of body mass). Primarily monounsaturated fatty acids with the exception of 18:1n ? 9, increased significantly from baseline by 12 h postprandial without greatly affecting total plasma lipid concentrations. Total plasma lipids then showed a small peak at 24 h postprandial, coinciding with a peak in triacylglycerols. We conclude that assimilation of lipids from the digest into the plasma is slower than reported for proteins and carbohydrates in the same species. Furthermore, as there were no significant differences between the HPV and DA, no measurable effect of hepatic passage on plasma lipid levels was resolved. Therefore, we also conclude that, in contrast to that in higher vertebrates, hepatic passage does not seem to have a major role in rainbow trout for modulating the postprandial plasma profile of lipids.     

Routine metabolism of larval green sturgeon (Acipenser medirostris Ayres)

Routine metabolic rates of green sturgeon (Acipenser medirostris) from hatching to 31 days post hatch (dph) were determined under normoxic conditions. During the endogenous feeding stage that comprised period from hatching to 15 dph, the oxygen consumption rate (MO₂, μg O₂ larva⁻¹ h⁻¹) increased 5-fold before yolk reserves became exhausted and MO₂ rates steady. The allometric relationships between MO₂ and body mass had mass exponents greater than 1.0 (b=1.64±0.21) and equal to 1.0 (b=1.04±0.07) during the endogenous and exogenous feeding phases, respectively. The magnitude and changes of MO₂ rates in green sturgeon larvae reflected their early ontogeny, especially during the endogenous feeding phase when the increase in metabolic rates was associated with organogenesis, acquisition of organ functions, and the conversion of yolk sac into new tissues. This revised version was published online in August 2006 with corrections to the Cover Date.     

Investigations on the digestibility and metabolizability of the major nutrients and energy of maize leaves and barnyard grass in grass carp (Ctenopharyngodon idella)

In the uplands of northern Vietnam, culture of grass carp contributes significantly to income and household food security of Black Thai farmers. Maize is one of the most important upland crops and barnyard grass is the most important weed in the paddy rice fields. Thus, these are frequently used by small‐scale fish farmers as fish feed. An 8‐week feeding trial was conducted simultaneously in a recirculation and in a respirometric system to determine the digestibility and metabolizability of the nutrients of maize leaves and barnyard grass, to assess their crude protein, lipid and energy conversion and to estimate the energy allocation in grass carp. The following diets were used: diet ‘A’ (reference diet) containing 39% crude protein with 19.8 MJ kg^?1 gross energy; diets ‘B’, ‘C’ and ‘D’, which contained the same amount of the reference diet as the control diet, supplemented with a known amount of dried barnyard grass, dried maize leaves or fresh maize leaves, respectively. Five fish were assigned to each treatment in each experimental unit. Reference and test diets were fed to fish and faecal samples were collected and the oxygen consumption was measured in order to set up an energy budget of the fish over the whole experimental period. The weekly development of the body weight was recorded. The body weight gain of the fish fed diet D was significantly higher than that of the group fed diet A, which also in turn was significantly higher than that of fish fed diet B or C. The apparent digestibility coefficients (ADC) of nutrients and gross energy for the different experimental diets in fish kept in the aquaria were, for diets A, B, C and D respectively, 94.1%, 60.9%, 70.5% and 84.7% for protein, 91.3%, 60.7%, 76.8% and 71.8% for lipid; 95.9%, 44.5%, 60.6% and 69.1% for gross energy. The partial ADC of plant leaf ingredients were determined and barnyard grass and dried maize leaves were found to be not only poorly digestible but having negative impact on the digestibility of the reference diet, while fresh maize leaves were well digested. The results of the present study indicated that fresh maize leaves have a good potential to be used as supplement in diets for grass carp.     

Short‐term physiological role of thyroxine administration on plasma metabolites and growth performance of Sterlet sturgeon Acipenser ruthenus

Thyroid hormones (THs) play important roles in regulating growth, development and physiological functions in vertebrates. To study the role of short‐term effects of thyroxine (T4) on metabolism and growth in female Sterlet sturgeon, thirty fish with a weight of 707.97 ± 37.15 g were divided into six tanks. Fish were injected intraperitoneal with (a) coconut oil alone (control), (b) 1 mg T4 kg per BW in coconut oil (T₁) and (c) 10 mg T4 kg per BW in coconut oil (T₁₀). Blood samples were collected at 0, 4, 7, 14, 28 and 60 days for plasma biochemical analysis, and body length and mass were determined at day 60. Based on results, higher growth was indicated in T₁₀ group. The highest T4 levels were observed on day 4 in the T₁ and T₁₀ treatments and gradually declined during the trial. Plasma T3 levels were highest on days 4 and 7 in the T₁ and T₁₀ groups, respectively. Plasma triglyceride levels were significantly elevated on day 28 in the T₁ group and on day 60 in the T₁₀ group. Plasma glucose concentrations were significantly elevated on days 7 and 14 in T₁₀ group. Overall, the present results suggest that a single injection of T4 can improve growth via changes in the metabolic profile of Sterlet sturgeon.     

不同水流速度对中华倒刺鲃幼鱼自由基代谢的影响

在(25±0.5) ℃下, 设置4个水流速度实验组, 分别为静水对照组0 bl/s(体长/秒)、1.0 bl/s组、2.0 bl/s组和4.0 bl/s组, 研究不同水流速度对中华倒刺鲃(Spinibarbus sinensis)幼鱼肌肉自由基代谢的影响, 训练周期为8周。结果显示, 肌肉超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-P_X)的活性随着水流速度的增加而升高, 训练组过氧化氢酶(CAT)活性出现不同程度的下降, 而抗氧化物谷胱甘肽(GSH)的含量未发生明显变化(P>0.05); 训练组肌肉总抗氧化能力(T-AOC)均高于对照组, 2.0 bl/s组最高, 4.0 bl/s组次之, 1.0 bl/s组最低, 且2.0 bl/s和4.0 bl/s组T-AOC与对照组有显著性差异(P<0.05)。8周训练后, 2.0 bl/s组丙二醛(MDA)含量显著低于其他3组(P<0.05), 而H₂O₂含量随着水流速度增加呈上升趋势。8周运动训练对骨骼肌B细胞淋巴瘤因子-2 (Bcl-2)、Bcl-2相关X蛋白(Bax)的含量及血清肌酸激酶(CK)活性无显著影响(P>0.05), 但2.0 bl/s组Bax和Bax/ Bcl-2显著低于4.0 bl/s组(P<0.05)。结果表明, 2.0 bl/s的水流速度可提高中华倒刺鲃肌肉的抗氧化能力, 降低脂质过氧化水平, 并在一定程度上抑制肌细胞凋亡。

     

Dietary n‐3 highly unsaturated fatty acids affect the biological and serum biochemical parameters,tissue fatty acid profile,antioxidation status and expression of lipid‐metabolism‐related genes in grass carp,Ctenopharyngodon idellus

A 95‐day feeding trial was conducted to determine the effects of n‐3 highly unsaturated fatty acids (n‐3 HUFA) on the growth, antioxidation and lipid metabolism in grass carp (Ctenopharyngodon idellus). Two isonitrogenous and isolipidic diets were formulated with either lard oil (LO) or fish oil (FO) as the main lipid source. The results showed that the intraperitoneal fat (IPF) ratio was significantly lower (P < 0.05) in FO group. The concentration of n‐3 HUFA in muscle, hepatopancreas and IPF was significantly higher in FO group (P < 0.05). The serum low‐density lipoprotein (LDL) content was significantly lower (P < 0.05), and glucose (GLU) content was significantly higher (P < 0.05) in FO group. The serum total superoxide dismutase (T‐SOD) activity was significantly higher (P < 0.05) in FO group, consistent with the serum malondialdehyde (MDA) content. The gene expression of IPF fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), sterol regulatory element‐binding protein (SREBP‐1) and peroxisome proliferator‐activated receptor γ (PPARγ) was significantly lower (P < 0.05) and that of peroxisome proliferator‐activated receptor α (PPARα) was significantly higher (P < 0.05) in FO group compared with LO group. Similar trends were found in the hepatopancreas, except for PPARγ. It is suggested that n‐3 HUFA could inhibit lipid accumulation in grass carp by affecting the expression of lipid‐metabolism‐related genes.