中草药淫羊藿和菟丝子对日本鳗鲡卵巢发育的影响
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摘要
在水温(21±1)℃下,给体重(592.5±52.5) g的日本鳗鲡(Anguilla japonica)雌亲鱼投喂每千克体重含0.034 g淫羊藿和0.034 g菟丝子浸膏的饲料90 d,对照组投喂人工配合饲料,研究淫羊藿和菟丝子对卵巢发育的影响。结果显示,饲料中添加淫羊藿和菟丝子的雌鱼卵巢大部分卵母细胞属第Ⅱ时相,性腺指数和肝体比均显著高于对照组(P<0.05);卵母细胞油滴明显增多,部分卵母细胞胞质已充满油滴,核仁变小增多;血清雌二醇(E2)和11-酮基睾酮(11-KT)水平显著升高(P<0.05),实验组11-KT含量约为对照组的4倍;肝脏卵黄蛋白原基因vtg表达量升高,实验组和对照组卵巢cyp19a1仅微量表达,肝脏未检测到erα和erβ的表达;实验组肌肉总脂肪酸、饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)和高度不饱和脂肪酸(HUFA)含量均显著高于对照组(P<0.05),最主要的高度不饱和脂肪酸花生四烯酸(AA)、二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)含量均显著高于对照组(P<0.05)。本研究表明,在亲鱼培育饲料中添加中草药淫羊藿和菟丝子促进了日本鳗鲡卵母细胞油滴和肝脏卵黄蛋白原增加,提高了肌肉高不饱和脂肪酸的积累,为卵黄生成和卵母细胞进一步发育做好更充分的准备。
In the present study, female Japanese eels(Anguilla japonica) with an average body weight of(592.5±52.5) g were fed with a commercial diet supplemented with two Chinese herbs Epimedium brevicornum Maxim. and Cuscuta chinensis Lam at a water temperature of(21±1)℃ for 90 d. Then, the histological structure of ovarian development; changes in serum 11-KT and E2 levels; mRNA expression levels of liver vitellogenin(vtg), estrogen receptors(erα, and erβ), and ovarian aromatase(cyp19 a1);, and the fatty acid composition were investigated by tissue section staining, immunoenzymatic assays, semi-quantitative PCR, and gas chromatography,respectively. The results showed that the gonadosomatic indices(GSIs) and the hepatosomatic indices(HISs) in the Japanese eels fed with diets containing E. brevicornum Maxim. and C. chinensis Lam. were significantly higher than those in the control group(P<0.05); the treated group also showed ovaries filled with advanced perinucleolar follicles and increased numbers of oil droplets, and significantly higher serum 11-KT and E2 levels(P<0.05). Expression of vtg mRNA was elevated in the liver of the treated group; however, cyp19 a1 mRNA levels were extremely low in the ovaries of both the control group and the treated group, and the expression of erα and erβ mRNA was not detected in the livers of the two groups. Total fatty acid levels, including saturated fatty acids(SFA), monounsaturated fatty acids(MUFA), and highly unsaturated fatty acids(HUFA), in muscle were significantly higher in the Japanese eels fed with diets containing E. brevicornum Maxim. and C. chinensis Lam than in the control group fish(P<0.05); particularly, the levels of arachidonic acid(AA), eicosapentaenoic acid(EPA),and docosahexaenoic acid(DHA) were higher in the treated group. These findings indicated that the two Chinese herbs, E. brevicornum Maxim. and C. chinensis Lam., induced early ovary development by increasing oil droplets of previtellogenic oocytes and promoted vitellogenin accumulation in the liver of parental female Japanese eels.Moreover, E. brevicornum Maxim. and C. chinensis Lam. accelerated absorption of fatty acids, particularly HUFA,from diets, and subsequently their accumulation in the muscle, to prepare for vitellogenesis and further development of oocytes.
In the present study, female Japanese eels(Anguilla japonica) with an average body weight of(592.5±52.5) g were fed with a commercial diet supplemented with two Chinese herbs Epimedium brevicornum Maxim. and Cuscuta chinensis Lam at a water temperature of(21±1)℃ for 90 d. Then, the histological structure of ovarian development; changes in serum 11-KT and E2 levels; mRNA expression levels of liver vitellogenin(vtg), estrogen receptors(erα, and erβ), and ovarian aromatase(cyp19 a1);, and the fatty acid composition were investigated by tissue section staining, immunoenzymatic assays, semi-quantitative PCR, and gas chromatography,respectively. The results showed that the gonadosomatic indices(GSIs) and the hepatosomatic indices(HISs) in the Japanese eels fed with diets containing E. brevicornum Maxim. and C. chinensis Lam. were significantly higher than those in the control group(P<0.05); the treated group also showed ovaries filled with advanced perinucleolar follicles and increased numbers of oil droplets, and significantly higher serum 11-KT and E2 levels(P<0.05). Expression of vtg mRNA was elevated in the liver of the treated group; however, cyp19 a1 mRNA levels were extremely low in the ovaries of both the control group and the treated group, and the expression of erα and erβ mRNA was not detected in the livers of the two groups. Total fatty acid levels, including saturated fatty acids(SFA), monounsaturated fatty acids(MUFA), and highly unsaturated fatty acids(HUFA), in muscle were significantly higher in the Japanese eels fed with diets containing E. brevicornum Maxim. and C. chinensis Lam than in the control group fish(P<0.05); particularly, the levels of arachidonic acid(AA), eicosapentaenoic acid(EPA),and docosahexaenoic acid(DHA) were higher in the treated group. These findings indicated that the two Chinese herbs, E. brevicornum Maxim. and C. chinensis Lam., induced early ovary development by increasing oil droplets of previtellogenic oocytes and promoted vitellogenin accumulation in the liver of parental female Japanese eels.Moreover, E. brevicornum Maxim. and C. chinensis Lam. accelerated absorption of fatty acids, particularly HUFA,from diets, and subsequently their accumulation in the muscle, to prepare for vitellogenesis and further development of oocytes.
引文
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[2]Okamura A,Horie N,Mikawa N,et al.Recent advances in artificial production of glass eels for conservation of anguillid eel populations[J].Ecology of Freshwater Fish,2013,23(1):5-110.
[3]Yamamoto J,Yamauchi K.Sexual maturation of Japanese eel and production of eel larvae in the aquarium[J].Nature,1974,251(5472):220-222.
[4]Motonobu T,Yamashita K,Oka H P.A memoir on the hatched larva of the Japanese eel,Anguilla japonica,from the adults matured artificially[J].Bulletin of the Shizuoka Prefectural Fisheries Experiment Station,1976,10:87-90.
[5]Satoh H,Yamamori K,Hibiya T.Induced spawning of the Japanese eel[J].Nippon Suisan Gakkaishi,1992,58(5):825-832.
[6]Ijiri S,Kayaba T,Takeda N,et al.Pretreatment reproductive stage and oocyte development induced by salmon pituitary homogenate in the Japanese eel Anguilla japonica[J].Fisheries Science,1998,64(4):531-537.
[7]Sato N,Kawazoe I,Suzuki Y,et al.Adequate interval of s GTH administration to induce vitellogenesis:and induction of ovulation by simultaneous administration of s GTHand17a-hydroxyprogesterone in the Japanese eel[J].Fisheries Science,2000,66(4):644-654.
[8]da Silva F F G,St?ttrupa J G,Kj?rsvik E,et al.Interactive effects of dietary composition and hormonal treatment on reproductive development of cultured female European eel,Anguilla anguilla[J].Animal Reproduction Science,2016,171:17-26.
[9]Izquierdo M S,Fernández-Palacios H,Tacon A G J.Effect of broodstock nutrition on reproductive performance of fish[J].Aquaculture,2001,197(1-4):25-42.
[10]Qiu X C,Zhou H Q,Liu X G,et al.The effect of dietary Chinese herb additives on growth and digestibility of allogynogenetic crucian carp[J].Journal of Fisheries of China,2002,26(6):551-555.[邱小琮,周洪琪,刘小刚,等.中草药添加剂对异育银鲫生长和蛋白质消化吸收的影响[J].水产学报,2002,26(6):551-555.]
[11]Wang J Q,Sun Y X,Zhang J C.Effects of Chinese herb additives on growth,digestive activity and non-specific immunity in flounder Paralichthys olivaceus[J].Journal of Fisheries of China,2006,30(1):90-96.[王吉桥,孙永新,张剑诚.金银花等复方草药对牙鲆生长、消化和免疫能力的影响[J].水产学报,2006,30(1):90-96.]
[12]Li Z Q,Guan R Z,Wang L H,et al.Antibacterial activity analysis for six kinds of Chinese herbs and their compounds against Pathogenic aeromonas from cultured eels in vitro[J].Acta Hydrobiologica Sinica,2012,36(1):85-92.[李忠琴,关瑞章,汪黎虹,等.六种中药及其复方对鳗鲡致病性气单胞菌的体外抑制作用[J].水生生物学报,2012,36(1):85-92.]
[13]Yan P X,Du B J,Luo R.Research progress in the similar effect of traditional Chinese medicine as hormones[J].China Journal of Traditional Chinese Medicine and Pharmacy,2014,29(2):531-534.[闫朋宣,杜宝俊,罗然.中药类激素样作用研究进展[J].中华中医药杂志,2014,29(2):531-534.]
[14]Li Y B,Meng F H,Lu X M,et al.Chemical constituents from herb of Epimedium brevicornum[J].China Journal of Chinese Materia Medica,2005,30(8):586-588.[李遇伯,孟繁浩,鹿秀梅,等.淫羊蕾化学成分的研究[J].中国中药杂志,2005,30(8):586-588.]
[15]Donnapee S,Li J,Yang X,et al.Cuscuta chinensis Lam.:Asystematic review on ethnopharmacology,phytochemistry and pharmacology of an important traditional herbal medicine[J].Journal of Ethnopharmacology,2014,157:292-308.
[16]Cornwell T,Cohick W,Raskin I.Dietary phytoestrogens and health[J].Phytochemistry,2004,65(8):995-1016.
[17]Williamson G,Barron D,Shimoi K.In vitro biological properties of flavonoid conjugates found in vivo[J].Free Radical Research,2005,39(5):457-469.
[18]Li F F,Li E,LüZ J,et al.Effects of icarrin on secretive function of cultured granulosa and adrenal cortical cells of rats[J].China Journal of Chinese Materia Medica,1997,22(8):499-500.[李芳芳,李恩,吕占军,等.淫羊藿甙对大鼠卵泡颗粒细胞和肾上腺皮质细胞分泌功能的影响[J].中国中药杂志,1997,22(8):499-500.]
[19]Yang L J,Lu D F,Guo J J,et al.Icariin from Epimedium brevicornum Maxim promotes the biosynthesis of estrogen by aromatase(CYP19)[J].Journal of Ethnopharmacology,2013,145(3):715-721.
[20]Zhang Z B,Yang Q T.The testosterone mimetic properties of icariin[J].Asian Journal of Andrology,2006,8(5):601-605.
[21]Yang J X,Wang Y L,Bao Y,et al.The total flavones from Semen cuscutae reverse the reduction of testosterone level and the expression of androgen receptor gene in kidney-yang deficient mice[J].Journal of Ethnopharmacology,2008,119(1):166-171.
[22]Yuan H,Cao S P,Chen S Y,et al.Research progress on chemical constituents and quality control of Epimedii folium[J].Chinese Traditional and Herbal Drugs,2014,45(24):3630-3640.[袁航,曹树萍,陈抒云,等.淫羊藿的化学成分及质量控制研究进展[J].中草药,2014,45(24):3630-3640.]
[23]Zheng H Z,Dong Z H,She J.Modern Study of Traditional Chinese Medicine(volume five)[M].Beijing:Xueyuan Press,1998:4110-4120.[郑虎占,董泽宏,佘靖.中药现代研究与应用(第五卷)[M].北京:学苑出版社,1998:4110-4120.]
[24]Yang B T,An F L,Liao J F,et al.Effects of herbal feed additives on the reproductive performance of ewes[J].Chinese Journal of Veterinary Drug,2010,44(10):51-54.[杨保田,安芳兰,廖吉方,等.中草药饲料添加剂对母羊繁殖性能的影响[J].中国兽药杂志,2010,44(10):51-54.]
[25]Wang J Q,Liu G Z,Tian Z X,et al.Effects of Chinese herb medicine supplementation on reproduction of yellow catfish Pelteobagrus fulvidraco[J].Journal of Dalian Fisheries University,2008,23(4):258-263.[王吉桥,柳圭泽,田忠信,等.中草药添加剂对黄颡鱼繁殖性能及效果的影响[J].大连水产学院学报,2008,23(4):258-263.]
[26]Zhao Q K,Ding W D,Bing X W,et al.The influence of Chinese herbal medicine feed additive on eel(Monopterus albus)sex hormone levels[J].Chinese Agricultural Science Bulletin,2014,30(32):38-42.[赵庆凯,丁炜东,邴旭文,等.中草药饲料添加剂对黄鳝性激素含量的影响[J].中国农学通报,2014,30(32):38-42.
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