高度不饱和脂肪酸对水生动物生长、发育和繁殖的影响与机理

许友卿，韩进华，陈亨德，钟艺文，丁兆坤

(广西大学 水产科学研究所，广西 南宁 535004)

高度不饱和脂肪酸是多不饱和脂肪酸中具有二十个碳原子以上，含3个或3个以上双键的脂肪酸[1-4]，主要有花生四烯酸(C20:4n-6)、二十二碳六烯酸(C22：6n-3)和二十碳五烯酸(C20:5n-3)等[5-12]。

高度不饱和脂肪酸影响机体脂类代谢[13]、基因表达[14-15]、细胞膜功能[16]、机体免疫[17-18]及血液生理生化特性[19-20]等，对水生动物的生长、发育和繁殖发挥重要的作用。然而，对高度不饱和脂肪酸的这种影响及机理研究较少，一些有关问题亟待探讨[21]。

笔者综述了高度不饱和脂肪酸对水生动物生长、发育和繁殖的影响及其作用机理，旨在掌握高度不饱和脂肪酸影响水生动物生长、发育和繁殖的规律和调控机理，有效地利用高度不饱和脂肪酸调控、促进水生动物的生长、发育和繁殖，提高生产效率，增加水产养殖业的经济效益和社会效益。

1 高度不饱和脂肪酸对水生动物繁殖的影响

高度不饱和脂肪酸能显著促进水生动物的生殖性能。高度不饱和脂肪酸如花生四烯酸、二十二碳六烯酸和二十碳五烯酸是水生动物合成卵黄和胚胎发育所必需，在水生动物性腺成熟过程中不仅作为一种能量来源，更能为性腺的连续发育和胚胎的形态发展提供必需营养素，包括必需脂肪酸、磷脂和某些激素。高度不饱和脂肪酸可维持水生动物亲体正常性成熟，促进卵黄正常发生和胚胎发育，提高卵的孵化率[22-23]。Xu等[24]发现，日粮中添加适当比例的n-6/n-3多不饱和脂肪酸促进鲤鱼(Cyprinuscarpio)性腺发育成熟、繁殖性能和雌鲤鱼产后恢复。鱼日粮中脂肪酸的组成对成功繁殖和后代存活至关重要[25-29]。多不饱和脂肪酸是影响亲鱼繁殖性能的关键营养素[30-31]。淡水、洄游鱼类通常需要C18多不饱和脂肪酸，而海洋鱼类则需要花生四烯酸、二十碳五烯酸和二十二碳六烯酸等高度不饱和脂肪酸[29,32]。这些重要的高度不饱和脂肪酸——花生四烯酸、二十碳五烯酸和二十二碳六烯酸，被极性脂质优先结合，在配子和胚胎发育中发挥特殊作用[33]。

饲料中添加多不饱和脂肪酸对促进鱼体成熟作用显著。真鲷(Pagrusmaior)的日粮如缺乏多不饱和脂肪酸，会显著降低孵化率，增加卵和仔鱼的畸形率[34-39]。事实证明，繁殖成功依赖于日粮中n-3和 n-6多不饱和脂肪酸的水平[37-41]，日粮中的n-3和 n-6多不饱和脂肪酸可以提高亲鱼的繁殖性能[37-38,42-44]。通常，受精卵含的n-3和 n-6多不饱和脂肪酸多为细胞膜磷脂，尤其是卵磷脂[45]。在鱼受精卵中，二十二碳六烯酸主要存在于磷脂酰胆碱中。在幼鱼发育过程中，二十二碳六烯酸被优先用于神经组织和视网膜的发育[46-47]。为亲鱼提供充足的n-3 和 n-6 多不饱和脂肪酸有利于胚胎和仔鱼的发育、细胞的增殖和分化[48-49]。在自身不能合成n-3和 n-6 多不饱和脂肪酸鱼类的日粮中必须添加n-3和 n-6多不饱和脂肪酸，如花生四烯酸、二十碳五烯酸和二十二碳六烯酸，或其前体，如亚油酸(LA,是花生四烯酸的前体物质)、亚麻酸(LNA,是二十碳五烯酸和二十二碳六烯酸的前体物质)[50-51]。投喂饵料鱼的Percafluviatilis和Sanderlucioperca亲鱼的受精卵和幼鱼质量高于投喂基于适宜磷脂和花生四烯酸、二十碳五烯酸和二十二碳六烯酸比例试验饲料的欧亚鲈鱼亲鱼[52]。但是，不同鱼类对多不饱和脂肪酸的需求不同，例如鲤鱼生长发育需要n-3和n-6多不饱和脂肪酸[53]，莫桑比克罗非鱼(Oreochromismossambicus)需要n-6 多不饱和脂肪酸，而虹鳟(Oncorhynchusmykiss)需要n-3 多不饱和脂肪酸[25]。在欧洲鲈鱼(Dicentrarchuslabrax)繁殖、生长和发育过程中，n-6高度不饱和脂肪酸发挥重要作用[54]。饲喂高水平花生四烯酸亲鱼的卵受精率和孵化率高于投喂低水平花生四烯酸的亲鱼卵[37]。在迈耶剑尾鱼(Xiphophorusmeyer)卵母细胞和鱼苗中沉积较多花生四烯酸，表明花生四烯酸在其生殖活动中的重要性[55]。

高度不饱和脂肪酸的含量及二十二碳六烯酸/二十碳五烯酸和花生四烯酸/二十二碳六烯酸比例显著影响水生动物的生殖性能及仔鱼质量。日粮中适量二十碳五烯酸和二十二碳六烯酸有利于水生动物吸收和运输胆固醇，为合成孕酮和雌二醇提供原料，促进孕酮和雌二醇的生成[56-57]。适宜的高度不饱和脂肪酸含量及其比例(如二十二碳六烯酸/二十碳五烯酸和花生四烯酸/二十二碳六烯酸等)会显著提高水生动物的生殖性能及幼体质量[60-61]。投喂富含n-3系列脂肪酸的饲料可以显著提高螯虾(Astacusleptodactylus)亲虾的产卵量和第一阶段幼虾的质量[62]。Buen-Ursua等[63]发现，二十二碳六烯酸/二十碳五烯酸比率提高海马(Hippocampuscomes)繁殖能力的作用，比其各自的水平高低更重要。Luo等[64]发现，饲喂高比例二十二碳六烯酸/二十碳五烯酸 (1.9∶1.0)的西伯利亚鲟(Acipenserbaeri)雌亲鱼的产卵质量、繁殖力、受精率分别比饲喂低比例二十二碳六烯酸/二十碳五烯酸 (1.0∶1.9)的雌亲鱼提高40.98%、22.3%、35.6%。Kohler[65]发现，白鲈鱼(Moronechrysops)卵含n-3高度不饱和脂肪酸包括二十碳五烯酸、二十二碳六烯酸和鲱油酸较高时，孵化率也高。Yanes-Roca等[66]报道，巴西黄金鲈(Centropomusundecimalis)含较高二十二碳六烯酸的受精卵孵化率较高。

高度不饱和脂肪酸不但可提高水生动物卵子的质量，还能增加精子质量和受精率。人们通常主要关心卵子质量，却未足够关注精子质量。实际上，精子质量同样影响亲本的繁育性能[67]。草鱼(Ctenopharynodonidellus)精子中的n-6多不饱和脂肪酸主要是花生四烯酸(含量13.95%)，显著高于卵子花生四烯酸含量(5.86%)[68]，表明花生四烯酸影响精子的活力[69]。高度不饱和脂肪酸对雄性硬骨鱼的性成熟和精子产生均发挥重要作用[70]。Asturiano等[71]用湿杂鱼和2种富含多不饱和脂肪酸的北半球鱼油、金枪鱼轨道油制备的颗粒商业饲料，分别投喂Dicentrarchuslabrax的亲鱼作比较，发现饲喂北半球鱼油和金枪鱼轨道油的雄亲鱼排精历时比饲喂湿杂鱼雄性亲鱼更长，排出的精液体积和精子密度都显著高于后者，但各组精子的质量和运动能力没有差异。尽管受精后3 h和24 h，它们的受精率相似，均在88%～90%，但是卵受精后48 h和72 h，投喂金枪鱼轨道油的胚胎和幼体成活率显著增高。于受精后48 h的胚胎和幼体成活率分别为：投喂北半球鱼油者13.9%，金枪鱼轨道油者20.9%，湿杂鱼者1.0%；于受精后72 h的胚胎和幼体成活率分别为，投喂北半球鱼油者15.5%，金枪鱼轨道油者20.6%，湿杂鱼者1.2%。

然而，Berenjestanaki等[72]报道，给三斑毛腹鱼(Trichopodustrichopterus)亲鱼投喂二十碳五烯酸、 二十二碳六烯酸等含量高的鱼油，对产卵的质量参数产生负面影响。

2 高度不饱和脂肪酸对水生动物发育的影响

高度不饱和脂肪酸对早期胚胎发育的影响很大。Araújo等[58]发现,摄食添加玉米油饲料的雌斑马鱼(Brachydaniorerio)亲鱼，其卵巢中花生四烯酸的比例(1.48 0.44%)(P= 0.015)高于饲喂其他饲料的雌鱼(P= 0.0069)，受精后8～9 h时受精卵发育较快，说明花生四烯酸可能是胚胎早期发育的调节剂，但是花生四烯酸的效应机制尚需研究。

高度不饱和脂肪酸对水生动物幼体的生长发育，尤其对骨骼发育及相关基因的表达影响显著。研究发现，投喂缺乏二十二碳六烯酸饲料的金头鲷(Sparusaurata)幼鱼体型较小，膀胱结石、脊椎前弯和后弯症的发病率高，而脊椎矿化的数量最少。增加饲料中二十二碳六烯酸的含量能增强幼鱼的生长，并显著提高类胰岛素生长因子-Ⅰ基因的表达[73]。然而，二十二碳六烯酸水平增至5%时，幼鱼组织脂质氧化程度增加，颅软骨内成骨、中轴骨骼的血液和椎弓畸形率增加，提高饲料中二十二碳六烯酸的水平，显著增加了氧化的风险，随之自由基和有毒的氧化混合物(脂肪酸过氧化物、脂肪酸羟基和醛)增加[74]，自由基和氧化反应产物可以引起哺乳动物骨细胞凋亡。更奇的是，提高饲料中二十二碳六烯酸水平时，也增加了幼鱼的氧化状态与幼鱼的骨骼畸形[73]。

然而，Hernández-Cruz等[75]报道，提高金头鲷饲料中二十二碳六烯酸的水平，既不影响骨骼畸形，也不影响骨标志(如与运行相关的转录因子2或碱性磷酸酶)基因的表达。

3 高度不饱和脂肪酸对水生动物生长的影响

n-3高度不饱和脂肪酸，尤其是二十二碳六烯酸和二十碳五烯酸，对于水生动物的生长和生理功能非常重要[76-80]。然而，有些水生动物不能合成或合成n-3 高度不饱和脂肪酸的能力有限，摄食是其获取n-3高度不饱和脂肪酸最有效和最主要的途径[81]。Hu等[82]研究发现，饲料中n-3高度不饱和脂肪酸水平显著影响三疣梭子蟹(Portunustrituberculatus)的质量增加量。但是，不同种类和比例的n-3高度不饱和脂肪酸影响相异，如二十二碳六烯酸对于促进仔鱼生长和发育效果比二十碳五烯酸效果更好[83-84]。用高比例二十二碳六烯酸/二十碳五烯酸 (1.9∶1.0)饲料投喂西伯利亚鲟雌鱼所孵化出35日龄稚鱼的体长较长，体质量较大，质量增加率及成活率较高，这些指标均比投喂低比例二十二碳六烯酸/二十碳五烯酸 (1.0∶1.9)的雌鱼所孵化的稚鱼更好[64]。

不同动物对n-3高度不饱和脂肪酸比例的响应相异。Xu 等[85]用6种不同二十二碳六烯酸/二十碳五烯酸(0.55、1.04、1.53、2.08、2.44)的饲料投喂日本尖吻鲈(Lateolabraxjaponicus)，发现饲料中二十二碳六烯酸/二十碳五烯酸比率在0.55～2.0时，鱼的终体质量和特定生长率随着二十二碳六烯酸/二十碳五烯酸比率的增加而显著增加，但比率高于2.0后则开始下降。比较用4种二十二碳六烯酸/二十碳五烯酸(0.70、0.84、1.06和 1.25)比例的饲料投喂三疣梭子蟹的效果，发现投喂二十二碳六烯酸/二十碳五烯酸为0.84的三疣梭子蟹终体质量和质量增加量最高，比投喂二十二碳六烯酸/二十碳五烯酸比例为1.06、1.25时显著提高，但与投喂二十二碳六烯酸/二十碳五烯酸为0.7的蟹比较，无显著差异[86]。Xu 等[87]发现，投喂不同二十二碳六烯酸/二十碳五烯酸比例的饲料时军曹鱼(Rachycentroncanadum)的生长无显著影响。

4 高度不饱和脂肪酸对水生动物生长、发育和繁殖的影响的机理

4.1 高度不饱和脂肪酸通过基因和受体影响水生动物生长、发育和繁殖

高度不饱和脂肪酸通过受体和基因表达而发挥作用。过氧化物酶体增殖物激活受体(PPARs)在介导高度不饱和脂肪酸影响代谢、生长、发育和繁殖中发挥重要作用。n-3 高度不饱和脂肪酸是过氧化物酶体增殖物激活受体的配体，过氧化物酶体增殖物激活受体可以调控大量与脂质代谢相关基因的表达。Kjr等[88]发现，二十二碳六烯酸可增强大西洋鲑(Salmosalar)过氧化物酶体增殖物激活受体α基因的表达。肝 X 受体是一种转录调控因子[89]。高度不饱和脂肪酸可调控肝 X 受体的表达[90]。

4.2 高度不饱和脂肪酸通过影响酶的表达、活性和激素来调控水生动物代谢、生长、发育和繁殖

高度不饱和脂肪酸可影响酶的表达及活性来调节水生动物代谢、生长、发育和繁殖。高度不饱和脂肪酸影响脂蛋白脂肪酶的表达及酶活性，进而影响机体脂肪代谢[91]及水生动物代谢、生长、发育和繁殖。研究表明，二十碳五烯酸能调节卵巢中雌二醇的生成[45]。饲料中含适量二十碳五烯酸和二十二碳六烯酸可促进孕酮和雌二醇的生成[56-57]。但过量的二十碳五烯酸和二十二碳六烯酸能够抑制金鱼类固醇类激素的合成，抑制卵黄合成[46,59]。

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