Regulation of adipocytes lipolysis by n-3 HUFA in grass carp (Ctenopharyngodon idellus) in vitro and in vivo

N-3 highly unsaturated fatty acids (n-3 HUFA) have been shown to inhibit body fat accumulation in animals. To clarify the mechanism of this fat-lowering effect of n-3 HUFA in grass carp (Ctenopharyngodon idellus), two experiments were conducted. In experiment 1, isolated grass carp mature adipocytes were incubated with docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) at different concentrations for 6 h. The release of glycerol to the medium was detected, and the expression of the lipolysis-related genes was analyzed. In experiment 2, a 95-day feeding trial was conducted with two diets formulated with either lard oil (as control) or fish oil (supplying n-3 HUFA as treatment) as the main lipid source. The glycerol and free fatty acid (FFA) released from the isolated adipocytes of both groups were detected after the feeding period. The expression of select lipolysis-related genes in adipose tissue was also analyzed. The results from experiment 1 showed that the release of glycerol was significantly increased by DHA and EPA (P < 0.05). Moreover, the expression of lipolysis-related genes, such as adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), tumor necrosis factor α (TNFα) and leptin, was also significantly elevated in the treatment group (P < 0.05). Experiment 2 demonstrated that glycerol and FFA release from the isolated adipocytes were significantly higher in the treatment group compared to the control group (P < 0.05). The expression level of ATGL, HSL, TNFα and leptin in the treatment group was significantly higher than in the control group (P < 0.05). The present results provide novel evidence that n-3 HUFAs could regulate grass carp adipocyte lipolysis in vitro or in vivo, and the effect might be in part associated with their influence on the expression of lipolysis-related genes and lipolysis-related adipokines genes.     

硫辛酸对草鱼脂肪细胞脂质含量及脂代谢相关基因表达的影响

为了探究硫辛酸对草鱼(Ctenopharyngodon idellus)脂肪细胞脂质代谢的影响,分离草鱼腹腔脂肪组织,消化得到脂肪细胞进行体外培养,分别用0、50和200μmol/L硫辛酸孵育细胞6 h,收取细胞和培养基样品后检测细胞甘油三酯(TG)含量、甘油和非酯化脂肪酸(NEFA)释放量及脂代谢相关基因表达。结果显示:50和200μmol/L硫辛酸处理显著降低草鱼脂肪细胞TG含量,200μmol/L硫辛酸显著增加细胞甘油和NEFA的释放量。200μmol/L硫辛酸组脂肪细胞脂肪分解相关基因(HSL、PPARα、CPT1、CD36)mRNA表达显著高于对照组,且ACC和PPARγ的mRNA表达水平显著上升,ATGL、MGL、SREBP-1c、FAS、DGAT等脂代谢基因表达状况无显著变化。综上所述,硫辛酸可能通过诱导草鱼脂肪细胞HSL和CPT1的基因表达促进脂质水解和β-氧化,从而降低脂肪细胞脂质含量。     

Short‐term starvation in silver pomfret (Pampus argenteus): molecular effects on lipid mobilization and utilization

To investigate the effects of short‐term starvation on lipid metabolism, juvenile silver pomfret (Pampus argenteus) with an initial weight of 18.1 ± 1.53 g were starved for 6 days. Fish were sampled on days 0, 2, 4 and 6 of starvation (S0, S2, S4 and S6, respectively) and then analysed. The results showed that short‐term starvation induced a significant decrease in visceral index, hepatosomatic index. The crude lipid content was also significantly decreased by 42.22% and 82.96% on S2 and S6 respectively. In addition, short‐term starvation significantly decreased the plasma triacylglycerol, non‐esterified fatty acid and low‐density lipoprotein cholesterol. Moreover, short‐term starvation significantly increased the expressions of hormone‐sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) at the mRNA and protein levels in the muscle and the ATGL expression at the protein level in the liver. The expressions of fatty acid translocase (CD36) and plasma membrane fatty acid‐binding protein (FABPpm) at the protein level in the muscle were significantly increased by short‐term starvation, while the expressions of fatty acid transport protein 1 (FATP1), FABPpm and lipoprotein lipase (LPL) at the protein level in the liver were significantly increased by short‐term starvation. Furthermore, the expressions of peroxisome proliferator activated receptor (PPAR) α and PPARγ at the mRNA and protein levels in the muscle were significantly elevated by short‐term starvation. These findings suggested that short‐term starvation increased lipid mobilization and utilization possibly through activation of lipolysis‐related genes (HSL and ATGL), lipid uptake‐related genes (LPL, CD36, FATP1 and FABPpm) and PPARs.     

Effect of waterborne zinc exposure on lipid deposition and metabolism in hepatopancreas and muscle of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

The aim of the present study was to explore the effect of waterborne zinc (control, 0.85, 2.20, 3.10 mg/l, respectively) exposure on lipid deposition and metabolism in the hepatopancreas and muscle of grass carp Ctenopharyngodon idella. The lipid content, Zn accumulation, and the activities and expression levels of several enzymes involved in lipid metabolism were determined in hepatopancreas and muscle. Waterborne Zn exposure reduced growth performance and increased Zn accumulation in both tested tissues. In hepatopancreas, Zn exposure increased lipid content, the activities of lipogenic enzymes, such as 6PGD, G6PD, ME, ICDH and FAS, as well as the mRNA expression level of G6PD, 6PGD, ICDH, FAS and SREBP-1. But the activity of CPT I and the mRNA expression of HSL, CPT Iα1a, CPT Iα2a and PPARα were down-regulated by Zn exposure. In contrast, in muscle, waterborne Zn exposure decreased lipid deposition, activities of 6GPD, ICDH and ME, as well as the mRNA expression level of G6PD, ICDH, ME, FAS and SREBP-1. However, the activity of CPT I as well as the mRNA expression level of PPARα, HSL, CPT Iα2a, CPT Iα1b and CPT Iβ were up-regulated by Zn exposure. Our results indicate that waterborne Zn increases lipid content by up-regulating lipogenesis and down-regulating lipolysis in hepatopancreas. But, in muscle, waterborne Zn reduces lipid accumulation by up-regulating lipolysis and down-regulating lipogenesis. Differential patterns of lipid deposition, enzymatic activities and genes’ expression indicate the tissue-specific regulatory mechanism in fish.     

Adrenergic Regulation of Lipid Mobilization in Fishes; a Possible Role in Hypoxia Survival

Lipids are for fish the major energy source. The nutritional conditions of most species vary throughout the year considerably. Thus storage and mobilization of lipids have to be tightly controlled, yet little is known about its control. Though several hormones are known to have a lipolytic effect, short term regulation of lipolysis is known for catecholamines only. Catecholamines are usually released under stress conditions, including hypoxia. In mammals these hormones have a strong lipolytic action, causing high plasma fatty acids levels during hypoxia. In contrast to mammals, several fish species show a decrease of plasma FFA-levels during hypoxia and anoxia. However, some studies gave contrasting results when catecholamines were administrated to different fish species. The reason for this may be due to opposing effects of catecholamines on lipolysis in different tissues. From catecholamine administration experiments in cannulated carp there is evidence that norepinephrine inhibits lipolysis via β₁- and β₃- adrenoceptors while β₂-adrenoceptors are involved in stimulation of lipolysis. Thus the opposite responses of different β-adrenoceptors may explain the conflicting in-vivo results obtained with fish. In vitro studies with adipocytes from different fish species confirm that activation of β-adrenoceptors suppresses lipolysis, while the opposite occurs in hepatocytes. Inhibiting β₁- and β₃- adrenoceptors in adipocytes were shown to be involved. Under hypoxia β-oxidation is inhibited, resulting in accumulation of fatty acids together with intermediates of the β-oxidation. This process may cause severe cellular damage in mammalian tissues, which ‘apparently’ does not occur in fishes. Fishes encounter (environmental) hypoxia on a regular basis, while for mammals hypoxia/anoxia is a pathological phenomenon. Hence the suppression of lipolysis in fishes under hypoxia (by β₁- and β₃- adrenoceptors) may be considered as a survival mechanism lost in higher vertebrates.     

慢性盐度胁迫下金钱鱼卵巢转录组分析

为了阐明盐度胁迫对金钱鱼(Scatophagus argus)代谢和生殖的影响,采用RNA-seq技术对低盐组(5)、对照组(25)和高盐组(35)处理40 d后的2龄性成熟金钱鱼卵巢进行转录组分析。结果发现,金钱鱼卵巢转录组测序获得raw reads共398681318,clean reads共396910398。从低盐胁迫相对对照组(5 vs 25)和高盐胁迫相对对照组(35 vs 25)中分别筛选到373个和874个差异表达基因(DEGs)。与对照组相比,低盐胁迫后氨基酸代谢相关基因(sds、bhmt)和脂肪酸代谢相关基因(pnpla2)显著下调;高盐胁迫后pgr、cyp17a1和ers1等生殖相关基因显著下调。KEGG通路富集分析发现,低盐胁迫组相对对照组显著富集与代谢相关的通路为半胱氨酸和甲硫氨酸代谢,缬氨酸、亮氨酸和异亮氨酸生物合成代谢,脂肪酸生物合成,脂肪酸代谢,皮质醇的合成和分泌,醛固酮的合成和分泌,脂肪细胞脂解的调节等;高盐胁迫组相对对照组中显著富集的与生殖相关的通路为雌激素信号传导通路。这些结果表明,氨基酸和脂肪酸的代谢调控可能在金钱鱼卵巢的低渗透压调节中起重要作用...     

Comparison of production efficiency and quality of differently cultured pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis> L.) juveniles as a valuable product for ongrowing culture

Production efficiency, frequency of fin erosion, skeletal and gill covers deformities, proximal composition of fish body with special emphasis on fatty acid contents in liver and muscle, oxidative stress and antioxidant response in gill, liver, muscle and intestine were analyzed and compared in exclusively RAS- and POND-cultured juveniles and juveniles produced with the combination of POND and RAS aquaculture in pikeperch (Sander lucioperca L.) with final body weight between 8.5 and 9.5 g. Higher production efficiency, lower fin erosion, no morphological deformities, higher level of essential fatty acids and lower oxidative stress were found in POND–RAS- and POND-cultured juveniles compared to exclusively RAS-produced pikeperch. This study showed that intensive pikeperch juvenile production still needs next technological innovations. On the other hand, the combination of POND–RAS aquaculture has provided a good potential to be an alternative production system producing high-quality juveniles for intensive or pond ongrowing culture mainly in countries with large pond area.     

Molecular cloning,characterization and nutritional regulation of key enzymes required for the effective utilization of marine wax esters by Atlantic salmon (Salmo salar L.)

Previous studies had shown that wax ester‐rich lipid extracted from calanoid copepods could be a useful alternative to fish oil as a provider of long‐chain n‐3 polyunsaturated fatty acids in diets for use in salmon aquaculture. Effective utilization of wax ester requires digestion and metabolism in the intestine with the fatty alcohol component being oxidized to fatty acid in intestinal cells through the combined activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). We studied wax ester utilization in Atlantic salmon using a candidate gene approach, focusing on ADH and ALDH as sequence information was available for these genes, including fish sequences, facilitating isolation of the cDNAs. Here, we report on the isolation and cloning of full‐length cDNAs for ADH3 and ALDH3a2 genes from salmon intestinal tissue. Functional characterization by heterologous expression in the yeast, Saccharomyces cerevisiae, showed the products of these cDNAs had long‐chain ADH and ALDH enzyme activities. Thus, ADH3 was capable of oxidizing long‐chain fatty alcohol, and ALDH3a2 was capable of oxidizing long‐chain fatty aldehyde to the corresponding fatty acid. The genes were highly expressed in intestinal tissue, particularly pyloric caeca, but their expression was not increased in salmon fed dietary copepod oil in comparison to fish fed fish oil.     

Habitat related variations in fatty acids of catadromous Galaxias maculatus

The fatty acid profile of the galaxiid fish Galaxias maculatus (Salmoniformes) collected from an estuarine habitat (a river mouth), a freshwater creek and two land-locked freshwater lakes in Victoria, Australia, were investigated with a view to evaluating habitat influences on the fatty acid profile. Fish from the freshwater creek were unpigmented, fresh-run juveniles (referred to as whitebait), the others adult fish. The fatty acids that predominated in adult G. maculatus muscle were 16:0 and 22:6(n-3), and 16:0 and 18:1(n-9), respectively. In whitebait 22:6(n-3), 20:5(n-3) and 16:0 were predominant. Hopkins estuary fish had the highest amount of 18:2(n-6) but the lowest 18:3(n-3). In all galaxiid populations, polyunsaturated fatty acids (PUFA) were the main group of fatty acids in total lipid in muscle, and accounted for more than 40 %, followed by saturates and monoenes. In landlocked populations, the amount of monoenes was lower than in other galaxiid populations. G. maculatus whitebait had significantly lower amount of saturates and the highest amount of n-3 fatty acids. The ratio of n-3 to n-6 PUFA ranged from 4.6 (Hopkins estuary) to 7.6 (whitebait), and the former differed significantly from all the other populations. The main fatty acids predominating in the liver of galaxiids from landlocked (Lake Bullen Merri) and estuarine populations were 16:0, 22:6(n-3) and 18:1 (n-9), and generally reflected the corresponding pattern in muscle. A principal component analysis of the fatty acid composition of muscle samples confirmed and highlighted the distinct fatty acid profile of the populations investigated, when three groupings could be discerned. The study suggests that in addition to the diet, other habitat related factors may influence the fatty acid profile in catadromous G. maculatus.     

Altered expression of CCAAT/enhancer binding protein and FABP11 genes during adipogenesis in vitro in Atlantic salmon (Salmo salar)

The study of CCAAT/enhancer binding proteins (C/EBPs) is important in the understanding of adipogenesis, but little is known about their regulation in fish. Here, we report three Atlantic salmon orthologs of c/ebp , and their expression in different tissues and in adipogenesis in vitro . During differentiation the expression of c/ebp α and fatp1 were upregulated in early differentiation stage with continuing high expression level in mature adipocytes, whereas c/ebp δ and fabp11 expression were elevated in mature adipocytes. Furthermore, the fatty acids eicosapentaenoic (EPA) and docosahexaenoic (DHA), suppressed the expression of the c/ebps, ppar β , and fatty acid transport protein ( fatp1 ) during terminal adipocyte differentiation. The study indicates that C/EBPs are induced upon the differentiation of primary-cultured adipocytes from Atlantic salmon and that marine n-3 highly unsaturated fatty acids (HUFAs) affect the c/ebp s expressions in mature adipocytes. Therefore, the established cell model described here appears to be valuable for studying modulation of fat content in farmed Atlantic salmon.     

The digestion and selective absorption of dietary fatty acids in Arctic charr, Salvelinus alpinus

To compare the rates of digestion and absorption of individual fatty acids, Arctic charr, Salvelinus alpinus (L.), were fed isoenergetic diets containing 40 g kg^?1 coconut oil, and various combinations of 10 g kg^?1 of polyunsaturated fatty acids (PUFA) (18:2n-6 or 18:3n-3) and monounsaturated fatty acids (MONO) (20:1n-9 or 22:1n-9) in the form of free fatty acids (FFA) or triacylglycerol (TAG). The average lipid digestibility for all diets measured by use of the chromic oxide method in the pyloric caeca area, midgut and hindgut were 72%, 83% and 88%, respectively, showing that lipid digestion and absorption occur mainly in the pyloric caeca area, but also extend throughout the intestinal tract. Analyses of digesta present in the intestinal segments suggest the predominance of non-specific lipolytic activity producing primarily FFA and glycerol from dietary TAG. Comparisons of the fatty acid composition of the lipid classes in the digesta showed that the utilization of dietary TAG was dependent both on the rate of release of the individual fatty acids from TAG, and their subsequent rate of absorption. When supplied as either FFA or TAG, the levels of PUFA (18:2n-6 or 18:3n-3) in the digesta were very low, indicating almost complete utilization. Both MONO used (20:1n-9 or 22:1n-9) were absorbed less efficiently than PUFA, but the rate of release from TAG seemed to be rate limiting only for 22:1n-9, which accumulated in the digesta. The rates of absorption of 20:1n-9 and 22:1n-9 when fed as FFA were the same. Comparisons of the levels of fatty acids in the dietary coconut oil TAG with those of the digesta lipids showed that 12:0 was a good substrate for intestinal lipase and was quickly absorbed. The lipolysis of 14:0 and 16:0 was intermediate while the longer-chain 18:0 appeared very resistant to digestion and was a major component of TAG, diacylglycerols and monoacylglycerols present in particularly the hindgut digesta. The absorption of 18:0 also appeared to be very low. The results suggest that PUFA are released very rapidly from dietary TAG by intestinal lipases in Arctic charr, and are specifically absorbed compared with long-chain saturated and monounsaturated fatty acids. The rate of lipolysis may be the rate-limiting step in the digestion of very long chain monounsaturated fatty acids such as 22:1n-9, while both the rate of lipolysis and absorption may be rate limiting for long-chain saturated fatty acids such as 18:0.     

A baseline study on lipid and fatty acid composition in the pearl oyster, <Emphasis Type="Italic">Pinctada fucata</Emphasis>

The pearl oyster Pinctada fucata is cultured for pearl production in China, but its germplasm is degenerated. Thus, lipid and fatty acids were investigated in P. fucata based on different tissues, body sizes and condition indexes using GC–MS to provide a lipidomic baseline for evaluation of breeding variety in terms of physiology. The major fatty acids over all samples included saturated (C16:0 and C18:0), monounsaturated [C16:1(n-7), C18:1(n-7) and C18:1(n-9)], and polyunsaturated fatty acids [arachidonic acid: C20:4(n-6), EPA: C20:5(n-3) and DHA: C22:6(n-3)]. Relative percentages of lipid contents of the mantle and visceral mass were, respectively, 40.75 and 42.03 %, significantly higher than adductor muscle (14.48 %) and gill (17.15 %). Fatty acid analysis showed that there were 15 types of fatty acids in adductor muscle, 17 types in mantle and gill, respectively, and 19 types in visceral mass. Meanwhile, total fatty acids, SFA, MUFA and PUFA were the highest in visceral mass (P < 0.05). Yet the lipid contents and fatty acid compositions showed no significant differences between body sizes or condition indexes based on visceral mass. The observations above suggested that visceral mass and mantle are the major tissues for P. fucata to store lipids and thus are important candidate tissues for lipidomic study.     

日本沼虾DEAD-box家族基因vasa和PL10的克隆及其在卵巢发育过程中的表达

vasa和PL10同属于DEAD-box基因家族,具有依赖ATP的RNA解螺旋酶活性。本研究中,克隆了日本沼虾DEAD-box家族的两个成员,分别为vasa(Mnv-asa)和PL10(MnP-L10)。其中Mn-vasa全长2 408bp,包含1 803 bp的开放阅读框,编码601个氨基酸;Mn-PL10全长2 607 bp,包含2 127 bp的开放阅读框,编码709个氨基酸。成体组织的组织检测表明,Mnv-asa仅在性腺中表达,而Mn-PL10在性腺和其他器官中均有表达。Mn-vasa和MnP-L10在卵巢的原位杂交结果显示:Mn-PL10和Mnv-asa都在卵黄发生前期,卵黄发生期的早期和中期表达,而在卵黄积累末期不表达,但Mn-PL10在卵黄发生中期定位于卵母细胞的两端,而Mn-vasa则是均匀的分布在卵母细胞细胞质中。     

饲料磷水平对吉富罗非鱼肠道磷和脂肪酸转运及甘油三酯沉积的影响

摘要:为了探究饲料中不同磷水平对罗非鱼肠道磷和脂肪酸转运及甘油三脂沉积的影响，我们配制了3组不同磷水平的饲料，饲料中磷含量分别为1.21 %（LPD）、1.75 %（MPD）和2.66 %（HPD）），用配制的饲料投喂罗非鱼10周。结果显示：与LPD组相比，HPD组显著增加罗非鱼肠道钠磷协同转运体SLC34A2基因（95.7%）和蛋白（200.1%）的表达，增加肠道组织总磷含量（79.2%）；HPD组上调脂肪酸吸收基因cd36、fabp1、fabp2和fatp1及甘油三酯合成基因dgat1的表达（11.2%~54.0%），下调脂肪酸转运基因apob-100的表达（37.7%），增加游离脂肪酸（NEFA）的含量（241.1%）。与LPD组相比，HPD组显著增加罗非鱼肠道组织甘油三酯（TG）的含量（42.9%）。 综上，我们的研究表明，饲料中高磷水平增加肠道组织TG的含量，上调脂肪酸吸收途径和抑制脂肪酸排出途径。我们的研究为深入揭示磷影响鱼类磷代谢及脂类代谢的机理奠定基础。     

缺刻缘绿藻FAD基因的序列特征及其相对转录量对氮饥饿的响应

为探讨缺刻缘绿藻(Myrmecia incisa)花生四烯酸(20:4 6,AA)合成过程中一系列脂肪酸去饱和酶(FAD)的作用,本研究克隆了12、6及5 FAD基因的cDNA全长序列(GenBank登录号分别为JN205757、JN205755和JN205756)及其相应的DNA序列。它们的cDNA全长分别为1 806 bp、2 674 bp及2 318 bp,其中ORF长为1 137bp、1 443 bp和1 446 bp,分别编码由378、480和481个氨基酸组成的蛋白;通过其cDNA与DNA序列的比对,发现12、6及5 FAD基因分别具有4、5和7个内含子,其剪切位点均符合"GT-AG"规则。Δ12、Δ6和Δ5 FAD编码蛋白均富含疏水性氨基酸,约占各自氨基酸组成的52.8%、46.6%及50.9%。密码子的偏好性与缺刻缘绿藻其他基因保持一致。推测这3个FAD基因编码蛋白都存在4个跨膜区,而12和5 FAD还各有一个明显不跨膜的疏水区;都具有FAD家族各自相对保守的3个组氨酸簇基序。基于缺刻缘绿藻和其他物种相应的FAD基因编码蛋白序列所构建的Neighbor-joining(NJ)系统进化树,表明Δ12、Δ6、Δ5及ω3 FAD分别隶属于各自的分支,其中,Δ12与ω3 FAD的亲缘关系较近,而Δ6与Δ5 FAD具有较近的亲缘关系。利用实时荧光定量PCR(quantitative real-time PCR,Q-RT-PCR)分析这3个FAD基因在缺刻缘绿藻缺氮培养96 h后又恢复氮培养72 h过程中的相对转录量,结果表明这3个基因的相对转录量都受到氮信号的调控,它们随着氮饥饿培养时间延长明显上升,而氮恢复培养后迅速并显著地下降到氮饥饿胁迫前的水平。结合已知脂肪酸成分在此过程中的变化,推测这3个基因对缺刻缘绿藻AA的合成和积累都起着重要的作用,而Δ6 FAD的作用可能更关键。     

The effect of ploidy on the fatty acid profile during the reproductive cycle of female rainbow trout (Oncorhynchus mykiss)

Rainbow trout Oncorhynchus mykiss triploids are regularly produced in fish farms to improve growth because the triploid females do not develop ovaries during the reproductive cycle. In this study, the tissue fatty acid allocations in triploid and diploid females were compared during the reproductive cycle to determine whether the ploidy influences the fatty acid profile of fish produced in aquaculture. The ovaries, liver, and white muscle fatty acid contents of diploid and triploid females were analyzed during the reproductive cycle. Diploid females tend to accumulate more polyunsaturated fatty acids than triploids during some phases of the reproductive cycle, and this profile was compensated by an increase in saturated and monounsaturated fatty acids in triploid females. Arachidonic acid (C20:4n6) was the main n6 polyunsaturated fatty acid in the ovaries of diploid females during the most advanced phases of the reproductive cycle, and docosahexaenoic acid (C22:6n3) was the main n3 polyunsaturated fatty acid. In triploid females, the percentage of both of these polyunsaturated fatty acids was lower than in diploid females during the most advanced phases of the reproductive cycle. In general, the lack of ovary development altered the hepatic synthesis of some fatty acids, mainly monounsaturated and polyunsaturated fatty acids, decreasing the content of the main fatty acids in the white muscle and, consequently, the mobilization of these specific fatty acids to the ovaries.     

斑马鱼营养性脂肪肝模型构建

近年来我国集约化水产养殖业发展势头强劲,而水产动物营养性疾病之鱼类脂肪肝的发病也逐渐迅猛起来,逐渐成为制约水产养殖业发展的因素之一,实验拟通过高脂饲料诱导建立斑马鱼营养性脂肪肝模型,为鱼类营养性脂肪肝相关研究工作的开展奠定基础。该实验分别以基础饲料和16%高脂饲料投喂1月龄斑马鱼,在投喂后第14、30、90天3个时间点取样,采用肝脏组织进行H.E染色、透射电镜、masson染色组织切片观察斑马鱼肝脏脂肪变性及纤维化程度;通过荧光定量PCR技术(q RT-PCR)检测斑马鱼肝脏脂合成及脂解相关基因:二酰甘油酰基转移酶基因(DGAT)、过氧化物酶体增殖物激活受体基因(PPAR)、胆固醇调节元件1基因(SREBP1)、脂肪酸合成酶基因(FAS)、诱导细胞死亡脱氧核糖核酸裂解因子样效应因子基因(CIDE)、激素敏感脂肪酶基因(HSL)、甘油三酯水解酶基因(ATGL)的表达水平;采用甲醇–氯仿法提取斑马鱼肝脏脂肪并检测肝脏中甘油三酯的含量。结果显示,在高脂饲料诱导第30天,光镜下可见发生弥漫性肝细胞脂肪变性,脂合成基因显著上调,甘油三脂的含量显著增加;诱导第90天,经透射电镜及masson染色组织切片观察结果证实,斑马鱼肝脏组织由于纤维化病变而使得肝组织功能受损,肝脏的脂合成基因及甘油三脂含量显著降低。研究表明,实验设计的高脂饲料对肝脏的损伤作用显著,30 d的投喂即可诱导发生脂肪肝,90 d投喂斑马鱼肝脏出现肝纤维化,此过程中肝脏脂合成基因表达量及甘油三酯含量先升高后降低,可知斑马鱼肝脏的损伤是由高脂饲料诱导的脂肪大量堆积而引起的,该营养性模型的成功构建在一定程度上弥补了以往构建斑马鱼高脂模型的缺陷,为研究鱼类营养性脂肪肝的发生机制提供了支撑。