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1 43 2 Vol.43, No OCEANOLOGIA ET LIMNOLOGIA SINICA Mar., 2012 (Plecoglossus altivelis) * ( ),,,,, 0h 10μm,,, 108h 120μm, 5 6, SOD POD MDA, 60h 36h 24h,, : 24h 48h 72h 96h 94.37% 88.19% 50.20% 10.83%, 85.78% 68.14% 25.54% 0.53%,,,,, Q492 (Plecoglossus altivelis) (Salmoniformes) (Osmeroidei) (Plecoglossidae) (Plecoglossus),,,, ( ), (, 1982),, 72h (, 1986),, (, 1995, 2009),,, 2h; 2 3h (, 1980);, 30 60min (, 1964;, 1989); 鲀, 4h (, 2005;, 1962), 鲀, (22 25 ), 8 10h, (15 ), 1d, 1, (Aegerter et al, 2004),, :,,,, * 973, 2008CB ;, IRT0734 ;, 2009C12077,,, limingyun@nbu.edu.cn : , :

2 海 314 洋与湖沼 43 卷方法 2009 年 10 月从浙江宁海凫溪香鱼养殖场挑选待 SOD 活力在缓慢升高, 但间隔 12h 的样品之间增幅不排卵亲鱼 50 余尾, 雌雄分养于网箱内, 适量投喂饲力与 0h 的卵细胞内酶活力相比升高 124%, 差异性显料每天 24h 不间断观察雌性香鱼腹部变化, 当有香著(P<0.05), 随后其活力开始下降, 72h 的活力比 60h 1.2 大, 差异性不显著(P>0.05) 在 60h 的时候 SOD 酶活鱼腹部膨大, 并轻压鱼腹有卵自流, 作为已经排卵标志, 然后将已经排卵亲鱼捞出暂养于专用网箱中, 共有 5 尾排卵亲鱼进行试验, 按照设定的时间分批次挤卵取样每次所获得卵分为两份, 一份经过人工干法受精后将受精卵用羽毛均匀黏附在铺于水中的洁净棕片上, 密度为 10 粒/cm2, 孵化水温为(19±1) (李明云等, 2003, 2009), 观察胚胎发育情况, 统计受精率和孵化率; 另一份用于卵细胞内油球等变化的观察和酶活力的测定, 计算不同时限所得卵子油球的比例, 并对油球变化进行拍照, 通过试剂盒(南京建成生物技术研究所)测定超氧化物歧化酶(SOD) 过氧化氢酶(POD)的活力以及丙二醛(MDA)含量, 试验步骤和方法按说明书进行 2 结果 2.1 卵内油球的变化第一次获得的适当成熟的卵母细胞卵内油球直径 10μm (图 1a) 12h 24h 36h 的细胞形态和色泽等特征并无太大变化, 其中 12h 24h 油球虽逐渐扩大(图 1b c d), 但仍呈颗粒状且分布均匀, 36h 卵内出现少数直径达 10μm 的油球图 1e 图 1f 是第 48h 和 72h 的卵球 48h 卵内油球直径为(20±10)μm, 以后油球继续变大, 并聚集融合 60h 卵内油球直径为(40±10)μm 72h 大部分油球直径可达 60μm, 少数较大的油球直径在 70μm 以上, 并逐渐向卵细胞植物极聚集(图 1g), 但在大油球的周围仍有较多的小油球随着保存时间的延长, 卵内油球体积继续增大, 84h 油球直径为(80±10)μm, 此时卵球内直径小于 20μm 的小油球已经很少, 在光镜下观察, 整个卵内部松散, 细胞内透明空间增多(图 1h) 当香鱼卵在腹腔中保存 96h 后, 卵内油球大部分融合成个大油球, 直径大都在 100μm 以上, 并集中在卵细胞的植物极一端, 细胞质浑浊不清晰 (图 1i) 108h 后, 此时的卵中在油球中会出现一个较大的油球, 位于植物极, 其直径大于 120μm (图 1j) 2.2 卵内酶活力和丙二醛含量的变化图 2 显示在香鱼卵子过熟的过程中, 超氧化物歧化酶(SOD)的活力呈现一个先升后降的趋势前 60h, 图1 香鱼卵细胞不同时限卵内油球的变化 Fig.1 Lipid droplet changes in over-aged ayu oocyte a. 排卵后 0h; b. 排卵后 12h; c. 排卵后 24h; d. 排卵后 36h; e. 排卵后 48h; f. 排卵后 60h; g. 排卵后 72h; h. 排卵后 84h; i. 排卵后 96h; j. 排卵后 108h FM: 附着膜, LD: 油球

3 2 : (Plecoglossus altivelis) %, (P<0.05) 36h, 0h 567%, 24 36h, (P<0.05);, 72h 2 (SOD) (POD) Fig.2 The activity of Superoxide dismutase (SOD) and Peroxidase (POD) during aging of post-ovulatory ayu oocyte 3 (MDA),, 24h, 0h MDA 186%, 12h, 36h, MDA 45%, MDA, MDA 24h, 36h MDA, (P<0.05) 3 (MDA) Fig.3 Content of Malondialdehyde (MDA) during aging of post-ovulatory ayu oocyte 2.3 1, 0h 95.01%,, 48h, 89% 60h, 75.61%,, 20%, 96h 10%;, 0h 90%, 12h 24h, 36h, 84.77%,, 60h, 50%, 72h, 25%, 72h, 10%, 96h ,,,,,,, (, 2009),,,,,, 95%, 90%,,,,,, 10 30μm,,, 90% 85%, 70%; 60h, 75.61%,, 20%,,,,,,,,,, 20% 5%; 96h,,,, 表 1 香鱼卵细胞的平均受精率和孵化率 Tab.1 Fertilization and hatching rate during aging of post-ovulatory ayu oocyte (h) (%) (%)

4 316 43,,, 10%, 0 Mansour (2007),,,, ; ;, 1 2 ; 1 2,, 0,,,, Lahnsteiner(2000, 2007),,, (Mansour et al, 2007) (Lahnsteiner, 2000),,,,, (, 1978),, 1/3 1/4,, 3.2 SOD, (2008) SOD, SOD,,, SOD POD, H 2 O 2, H 2 O 2, (, 2000) (2010) POD, POD,,, POD MDA, (polyunsaturated fatty acid, PUFA),,, (, 2010;, 2011),, SOD POD, MDA, MDA SOD POD,, SOD POD, 24h MDA, SOD POD MDA,, MDA,,,,,, 0h POD, SOD,, MDA, POD, SOD ; SOD POD, MDA, MDA, POD, SOD SOD,, POD, MDA POD (, 2010)

5 2 : (Plecoglossus altivelis) 317,,, AST ALT ALP NO SOD MDA., 21(1): 9 11, , 8: 1 33,, , 10: 1 7, (Mugil so-ouy Basilewsky). ( ), (3): 75 84,, , 33(4): 30 32,,, , 13: 309,,, , 16(6): 13 16,,, , 17(1): 18 20, 鲀., 35(2): 21 22, , 2: 19 21,,, , 22(5): ,, , 19(5): ,,, Hg 2+ (Eriocheir sinensis)., 42(3): ,, ( ), 24(3): 91 94,, , 6(2): ,,, , 3: Aegerter S, Jalabert B, Effects of post-ovulatory oocyte aging and temperature on egg quality and on the occurrence of triploid fry in rainbow trout, Oncorhynchus mykiss. Aquaculture, 231: Lahnsteiner F, First results on a relation between ovarian fluid and egg proteins of Salmo trutta and egg quality. Aquaculture Research, 38(2): Lahnsteiner F, Morphological, physiological and biochemical parameters characterizing the over-ripening of rainbow trout eggs. Fish Physiology and Biochemistry, 23(2): Mansour N, Lahnsteiner F, Patzer R A, Distribution of lipid droplets is an indicator of egg quality in brown trout, Salmo trutta fario. Aquaculture, 273: VARIATIONS OF PLECOGLOSSUS ALTIVELIS OOCYTE DURING POST-OVULATORY AGING ON THE LIPID DROPLET, ENZYME ACTIVITY, MDA, FERTILIZATION AND HATCHING RATES LI Ming-Yun, MIAO Liang, AN Qin, WANG Cheng-Yi, TANG Xian-Nian, CHEN Jiong, SHI Yu-Hong (Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo, ) Abstract Ayu Plecoglossus altivelis is a migrational fish without mesovarium. In the abdominal cavity, the oocyte is protected by the abdominal cavity fluid which can store it for a long time with a high fertilization potential. During this period the quality of the oocyte becomes low, choice of the maturation of adult ayu was based on the acquisition of the oocyte and sperm such that for the same female fish the acquisition of oocyte repeated per 12 hours. A part of each oocyte batch which was used for fertilization remained for observation. Through observation it was expected that there would be some differences between different egg batches including oocyte morphology, color and morphology of the lipid droplet. As the time that eggs stored in the abdominal cavity increased, the oocyte quality was lowered and this was reflected by the diameter of the lipid droplet becoming 10μm to 120μm from 0h to 96h and the amount of the lipid droplet only being five or six at 108h. The activity of SOD, POD and the content of MDA increased at the beginning then descended, and they exhibited the highest level at 60, 36 and 24 hours. With the variations of the lipid droplet and enzyme activity, egg quality declined gradually. Thus, while 94.37% fertilization rate and 85.78% hatching rate were observed, in average, at 24h, fertilization and hatching rate dropped to 88.19% and 68.14% at 48h, to 50.20% and 25.54% at 72h, and to 10.83% and 0.53% at 96h. Key words Ayu Plecoglossus altivelis, Oocyte, Lipid droplet, Enzyme activity, Fertilization rate, Hatching rate

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗 1338 中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月九味肝泰胶囊对高脂饮食诱导大鼠非酒精性脂肪肝的治疗作用陈菲 1 2,, 艾国 3, 盛柳青 1, 麻佳蕾 1, 黄正明 1. 金华职业技术学院医学院, 浙江金华