高度不飽和脂肪酸對水生動(dòng)物生長、發(fā)育和繁殖的影響與機(jī)理

許友卿，韓進(jìn)華，陳亨德，鐘藝文，丁兆坤

(廣西大學(xué) 水產(chǎn)科學(xué)研究所，廣西 南寧 535004)

高度不飽和脂肪酸是多不飽和脂肪酸中具有二十個(gè)碳原子以上，含3個(gè)或3個(gè)以上雙鍵的脂肪酸[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]。

高度不飽和脂肪酸影響機(jī)體脂類代謝[13]、基因表達(dá)[14-15]、細(xì)胞膜功能[16]、機(jī)體免疫[17-18]及血液生理生化特性[19-20]等，對水生動(dòng)物的生長、發(fā)育和繁殖發(fā)揮重要的作用。然而，對高度不飽和脂肪酸的這種影響及機(jī)理研究較少，一些有關(guān)問題亟待探討[21]。

筆者綜述了高度不飽和脂肪酸對水生動(dòng)物生長、發(fā)育和繁殖的影響及其作用機(jī)理，旨在掌握高度不飽和脂肪酸影響水生動(dòng)物生長、發(fā)育和繁殖的規(guī)律和調(diào)控機(jī)理，有效地利用高度不飽和脂肪酸調(diào)控、促進(jìn)水生動(dòng)物的生長、發(fā)育和繁殖，提高生產(chǎn)效率，增加水產(chǎn)養(yǎng)殖業(yè)的經(jīng)濟(jì)效益和社會(huì)效益。

1 高度不飽和脂肪酸對水生動(dòng)物繁殖的影響

高度不飽和脂肪酸能顯著促進(jìn)水生動(dòng)物的生殖性能。高度不飽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動(dòng)物合成卵黃和胚胎發(fā)育所必需，在水生動(dòng)物性腺成熟過程中不僅作為一種能量來源，更能為性腺的連續(xù)發(fā)育和胚胎的形態(tài)發(fā)展提供必需營養(yǎng)素，包括必需脂肪酸、磷脂和某些激素。高度不飽和脂肪酸可維持水生動(dòng)物親體正常性成熟，促進(jìn)卵黃正常發(fā)生和胚胎發(fā)育，提高卵的孵化率[22-23]。Xu等[24]發(fā)現(xiàn)，日糧中添加適當(dāng)比例的n-6/n-3多不飽和脂肪酸促進(jìn)鯉魚(Cyprinuscarpio)性腺發(fā)育成熟、繁殖性能和雌鯉魚產(chǎn)后恢復(fù)。魚日糧中脂肪酸的組成對成功繁殖和后代存活至關(guān)重要[25-29]。多不飽和脂肪酸是影響親魚繁殖性能的關(guān)鍵營養(yǎng)素[30-31]。淡水、洄游魚類通常需要C18多不飽和脂肪酸，而海洋魚類則需要花生四烯酸、二十碳五烯酸和二十二碳六烯酸等高度不飽和脂肪酸[29,32]。這些重要的高度不飽和脂肪酸——花生四烯酸、二十碳五烯酸和二十二碳六烯酸，被極性脂質(zhì)優(yōu)先結(jié)合，在配子和胚胎發(fā)育中發(fā)揮特殊作用[33]。

飼料中添加多不飽和脂肪酸對促進(jìn)魚體成熟作用顯著。真鯛(Pagrusmaior)的日糧如缺乏多不飽和脂肪酸，會(huì)顯著降低孵化率，增加卵和仔魚的畸形率[34-39]。事實(shí)證明，繁殖成功依賴于日糧中n-3和 n-6多不飽和脂肪酸的水平[37-41]，日糧中的n-3和 n-6多不飽和脂肪酸可以提高親魚的繁殖性能[37-38,42-44]。通常，受精卵含的n-3和 n-6多不飽和脂肪酸多為細(xì)胞膜磷脂，尤其是卵磷脂[45]。在魚受精卵中，二十二碳六烯酸主要存在于磷脂酰膽堿中。在幼魚發(fā)育過程中，二十二碳六烯酸被優(yōu)先用于神經(jīng)組織和視網(wǎng)膜的發(fā)育[46-47]。為親魚提供充足的n-3 和 n-6 多不飽和脂肪酸有利于胚胎和仔魚的發(fā)育、細(xì)胞的增殖和分化[48-49]。在自身不能合成n-3和 n-6 多不飽和脂肪酸魚類的日糧中必須添加n-3和 n-6多不飽和脂肪酸，如花生四烯酸、二十碳五烯酸和二十二碳六烯酸，或其前體，如亞油酸(LA,是花生四烯酸的前體物質(zhì))、亞麻酸(LNA,是二十碳五烯酸和二十二碳六烯酸的前體物質(zhì))[50-51]。投喂餌料魚的Percafluviatilis和Sanderlucioperca親魚的受精卵和幼魚質(zhì)量高于投喂基于適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試驗(yàn)飼料的歐亞鱸魚親魚[52]。但是，不同魚類對多不飽和脂肪酸的需求不同，例如鯉魚生長發(fā)育需要n-3和n-6多不飽和脂肪酸[53]，莫桑比克羅非魚(Oreochromismossambicus)需要n-6 多不飽和脂肪酸，而虹鱒(Oncorhynchusmykiss)需要n-3 多不飽和脂肪酸[25]。在歐洲鱸魚(Dicentrarchuslabrax)繁殖、生長和發(fā)育過程中，n-6高度不飽和脂肪酸發(fā)揮重要作用[54]。飼喂高水平花生四烯酸親魚的卵受精率和孵化率高于投喂低水平花生四烯酸的親魚卵[37]。在邁耶劍尾魚(Xiphophorusmeyer)卵母細(xì)胞和魚苗中沉積較多花生四烯酸，表明花生四烯酸在其生殖活動(dòng)中的重要性[55]。

高度不飽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顯著影響水生動(dòng)物的生殖性能及仔魚質(zhì)量。日糧中適量二十碳五烯酸和二十二碳六烯酸有利于水生動(dòng)物吸收和運(yùn)輸膽固醇，為合成孕酮和雌二醇提供原料，促進(jìn)孕酮和雌二醇的生成[56-57]。適宜的高度不飽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)會(huì)顯著提高水生動(dòng)物的生殖性能及幼體質(zhì)量[60-61]。投喂富含n-3系列脂肪酸的飼料可以顯著提高螯蝦(Astacusleptodactylus)親蝦的產(chǎn)卵量和第一階段幼蝦的質(zhì)量[62]。Buen-Ursua等[63]發(fā)現(xiàn)，二十二碳六烯酸/二十碳五烯酸比率提高海馬(Hippocampuscomes)繁殖能力的作用，比其各自的水平高低更重要。Luo等[64]發(fā)現(xiàn)，飼喂高比例二十二碳六烯酸/二十碳五烯酸 (1.9∶1.0)的西伯利亞鱘(Acipenserbaeri)雌親魚的產(chǎn)卵質(zhì)量、繁殖力、受精率分別比飼喂低比例二十二碳六烯酸/二十碳五烯酸 (1.0∶1.9)的雌親魚提高40.98%、22.3%、35.6%。Kohler[65]發(fā)現(xiàn)，白鱸魚(Moronechrysops)卵含n-3高度不飽和脂肪酸包括二十碳五烯酸、二十二碳六烯酸和鯡油酸較高時(shí)，孵化率也高。Yanes-Roca等[66]報(bào)道，巴西黃金鱸(Centropomusundecimalis)含較高二十二碳六烯酸的受精卵孵化率較高。

高度不飽和脂肪酸不但可提高水生動(dòng)物卵子的質(zhì)量，還能增加精子質(zhì)量和受精率。人們通常主要關(guān)心卵子質(zhì)量，卻未足夠關(guān)注精子質(zhì)量。實(shí)際上，精子質(zhì)量同樣影響親本的繁育性能[67]。草魚(Ctenopharynodonidellus)精子中的n-6多不飽和脂肪酸主要是花生四烯酸(含量13.95%)，顯著高于卵子花生四烯酸含量(5.86%)[68]，表明花生四烯酸影響精子的活力[69]。高度不飽和脂肪酸對雄性硬骨魚的性成熟和精子產(chǎn)生均發(fā)揮重要作用[70]。Asturiano等[71]用濕雜魚和2種富含多不飽和脂肪酸的北半球魚油、金槍魚軌道油制備的顆粒商業(yè)飼料，分別投喂Dicentrarchuslabrax的親魚作比較，發(fā)現(xiàn)飼喂北半球魚油和金槍魚軌道油的雄親魚排精歷時(shí)比飼喂?jié)耠s魚雄性親魚更長，排出的精液體積和精子密度都顯著高于后者，但各組精子的質(zhì)量和運(yùn)動(dòng)能力沒有差異。盡管受精后3 h和24 h，它們的受精率相似，均在88%～90%，但是卵受精后48 h和72 h，投喂金槍魚軌道油的胚胎和幼體成活率顯著增高。于受精后48 h的胚胎和幼體成活率分別為：投喂北半球魚油者13.9%，金槍魚軌道油者20.9%，濕雜魚者1.0%；于受精后72 h的胚胎和幼體成活率分別為，投喂北半球魚油者15.5%，金槍魚軌道油者20.6%，濕雜魚者1.2%。

然而，Berenjestanaki等[72]報(bào)道，給三斑毛腹魚(Trichopodustrichopterus)親魚投喂二十碳五烯酸、 二十二碳六烯酸等含量高的魚油，對產(chǎn)卵的質(zhì)量參數(shù)產(chǎn)生負(fù)面影響。

2 高度不飽和脂肪酸對水生動(dòng)物發(fā)育的影響

高度不飽和脂肪酸對早期胚胎發(fā)育的影響很大。Araújo等[58]發(fā)現(xiàn),攝食添加玉米油飼料的雌斑馬魚(Brachydaniorerio)親魚，其卵巢中花生四烯酸的比例(1.48 0.44%)(P= 0.015)高于飼喂其他飼料的雌魚(P= 0.0069)，受精后8～9 h時(shí)受精卵發(fā)育較快，說明花生四烯酸可能是胚胎早期發(fā)育的調(diào)節(jié)劑，但是花生四烯酸的效應(yīng)機(jī)制尚需研究。

高度不飽和脂肪酸對水生動(dòng)物幼體的生長發(fā)育，尤其對骨骼發(fā)育及相關(guān)基因的表達(dá)影響顯著。研究發(fā)現(xiàn)，投喂缺乏二十二碳六烯酸飼料的金頭鯛(Sparusaurata)幼魚體型較小，膀胱結(jié)石、脊椎前彎和后彎癥的發(fā)病率高，而脊椎礦化的數(shù)量最少。增加飼料中二十二碳六烯酸的含量能增強(qiáng)幼魚的生長，并顯著提高類胰島素生長因子-Ⅰ基因的表達(dá)[73]。然而，二十二碳六烯酸水平增至5%時(shí)，幼魚組織脂質(zhì)氧化程度增加，顱軟骨內(nèi)成骨、中軸骨骼的血液和椎弓畸形率增加，提高飼料中二十二碳六烯酸的水平，顯著增加了氧化的風(fēng)險(xiǎn)，隨之自由基和有毒的氧化混合物(脂肪酸過氧化物、脂肪酸羥基和醛)增加[74]，自由基和氧化反應(yīng)產(chǎn)物可以引起哺乳動(dòng)物骨細(xì)胞凋亡。更奇的是，提高飼料中二十二碳六烯酸水平時(shí)，也增加了幼魚的氧化狀態(tài)與幼魚的骨骼畸形[73]。

然而，Hernández-Cruz等[75]報(bào)道，提高金頭鯛飼料中二十二碳六烯酸的水平，既不影響骨骼畸形，也不影響骨標(biāo)志(如與運(yùn)行相關(guān)的轉(zhuǎn)錄因子2或堿性磷酸酶)基因的表達(dá)。

3 高度不飽和脂肪酸對水生動(dòng)物生長的影響

n-3高度不飽和脂肪酸，尤其是二十二碳六烯酸和二十碳五烯酸，對于水生動(dòng)物的生長和生理功能非常重要[76-80]。然而，有些水生動(dòng)物不能合成或合成n-3 高度不飽和脂肪酸的能力有限，攝食是其獲取n-3高度不飽和脂肪酸最有效和最主要的途徑[81]。Hu等[82]研究發(fā)現(xiàn)，飼料中n-3高度不飽和脂肪酸水平顯著影響三疣梭子蟹(Portunustrituberculatus)的質(zhì)量增加量。但是，不同種類和比例的n-3高度不飽和脂肪酸影響相異，如二十二碳六烯酸對于促進(jìn)仔魚生長和發(fā)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)飼料投喂西伯利亞鱘雌魚所孵化出35日齡稚魚的體長較長，體質(zhì)量較大，質(zhì)量增加率及成活率較高，這些指標(biāo)均比投喂低比例二十二碳六烯酸/二十碳五烯酸 (1.0∶1.9)的雌魚所孵化的稚魚更好[64]。

不同動(dòng)物對n-3高度不飽和脂肪酸比例的響應(yīng)相異。Xu 等[85]用6種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)的飼料投喂日本尖吻鱸(Lateolabraxjaponicus)，發(fā)現(xiàn)飼料中二十二碳六烯酸/二十碳五烯酸比率在0.55～2.0時(shí)，魚的終體質(zhì)量和特定生長率隨著二十二碳六烯酸/二十碳五烯酸比率的增加而顯著增加，但比率高于2.0后則開始下降。比較用4種二十二碳六烯酸/二十碳五烯酸(0.70、0.84、1.06和 1.25)比例的飼料投喂三疣梭子蟹的效果，發(fā)現(xiàn)投喂二十二碳六烯酸/二十碳五烯酸為0.84的三疣梭子蟹終體質(zhì)量和質(zhì)量增加量最高，比投喂二十二碳六烯酸/二十碳五烯酸比例為1.06、1.25時(shí)顯著提高，但與投喂二十二碳六烯酸/二十碳五烯酸為0.7的蟹比較，無顯著差異[86]。Xu 等[87]發(fā)現(xiàn)，投喂不同二十二碳六烯酸/二十碳五烯酸比例的飼料時(shí)軍曹魚(Rachycentroncanadum)的生長無顯著影響。

4 高度不飽和脂肪酸對水生動(dòng)物生長、發(fā)育和繁殖的影響的機(jī)理

4.1 高度不飽和脂肪酸通過基因和受體影響水生動(dòng)物生長、發(fā)育和繁殖

高度不飽和脂肪酸通過受體和基因表達(dá)而發(fā)揮作用。過氧化物酶體增殖物激活受體(PPARs)在介導(dǎo)高度不飽和脂肪酸影響代謝、生長、發(fā)育和繁殖中發(fā)揮重要作用。n-3 高度不飽和脂肪酸是過氧化物酶體增殖物激活受體的配體，過氧化物酶體增殖物激活受體可以調(diào)控大量與脂質(zhì)代謝相關(guān)基因的表達(dá)。Kjr等[88]發(fā)現(xiàn)，二十二碳六烯酸可增強(qiáng)大西洋鮭(Salmosalar)過氧化物酶體增殖物激活受體α基因的表達(dá)。肝 X 受體是一種轉(zhuǎn)錄調(diào)控因子[89]。高度不飽和脂肪酸可調(diào)控肝 X 受體的表達(dá)[90]。

4.2 高度不飽和脂肪酸通過影響酶的表達(dá)、活性和激素來調(diào)控水生動(dòng)物代謝、生長、發(fā)育和繁殖

高度不飽和脂肪酸可影響酶的表達(dá)及活性來調(diào)節(jié)水生動(dòng)物代謝、生長、發(fā)育和繁殖。高度不飽和脂肪酸影響脂蛋白脂肪酶的表達(dá)及酶活性，進(jìn)而影響機(jī)體脂肪代謝[91]及水生動(dòng)物代謝、生長、發(fā)育和繁殖。研究表明，二十碳五烯酸能調(diào)節(jié)卵巢中雌二醇的生成[45]。飼料中含適量二十碳五烯酸和二十二碳六烯酸可促進(jìn)孕酮和雌二醇的生成[56-57]。但過量的二十碳五烯酸和二十二碳六烯酸能夠抑制金魚類固醇類激素的合成，抑制卵黃合成[46,59]。

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