Wu Chenglong, Wang Jia, Xu Wei, Zhang Wenbing, Mai Kangsen
The Key Laboratory of Mariculture (Education Ministry of China), Ocean University of China, 5 Yushan Road, Qingdao, Shandong 266003, PR China; Zhejiang Provincial Key Laboratory of Aquatic Resources Conservation and Development, Huzhou University, 1 Xueshi Road, Huzhou 313000, PR China.
The Key Laboratory of Mariculture (Education Ministry of China), Ocean University of China, 5 Yushan Road, Qingdao, Shandong 266003, PR China.
Fish Shellfish Immunol. 2014 Dec;41(2):120-5. doi: 10.1016/j.fsi.2014.08.026. Epub 2014 Sep 1.
This study was conducted to investigate the effects of dietary ascorbic acid (AA) on transcriptional expression patterns of antioxidant proteins, heat shock proteins (HSP) and nuclear factor kappa B (NF-κB) in the hepatopancreas of Pacific abalone Haliotis discus hannai Ino (initial average length: 84.36 ± 0.24 mm) using real-time quantitative PCR assays. L-ascorbyl-2-molyphosphate (LAMP) was added to the basal diet to formulate four experimental diets containing 0.0, 70.3, 829.8 and 4967.5 mg AA equivalent kg(-1) diets, respectively. Each diet was fed to triplicate groups of adult abalone in acrylic tanks (200 L) in a flow-through seawater system. Each tank was stocked with 15 abalone. Animals were fed once daily (17:00) to apparent satiation for 24 weeks. The results showed that the dietary AA (70.3 mg kg(-1)) could significantly up-regulate the expression levels of Cu/Zn superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), feritin (FT) and heat shock protein 26 (HSP26) in the hepatopancreas of abalone in this treatment compared to the controls. However, the expression levels of Mn-SOD, glutathione peroxidase (GPX), thioredoxin peroxidase (TPx), selenium-binding protein (SEBP), HSP70 and HSP90 were significantly down-regulated. Compared with those in the group with 70.3 mg kg(-1) dietary AA, the expression levels of CAT, GST and HSP26 were decreased in abalone fed with very high dietary AA (4967.5 mg kg(-1)). In addition, significant up-regulations of expression levels of Mn-SOD, GPX, TPx, SEBP, FT, HSP70, HSP90 and NF-κB were observed in abalone fed with apparently excessive dietary AA (829.8 and 4967.5 mg kg(-1)) as compared to those fed 70.3 mg kg(-1) dietary AA. These findings showed that dietary AA influenced the expression levels of antioxidant proteins, heat shock proteins and NF-κB in the hepatopancreas of abalone at transcriptional level. Levels of dietary AA that appeared adequate (70.3 mg kg(-1)) reduced the oxidative stress by influencing gene expression of antioxidant proteins, but excessive dietary AA (829.8 and 4967.5 mg kg(-1)) induced oxidative stress in Pacific abalone H. discus hannai.
本研究旨在通过实时定量PCR分析,研究日粮抗坏血酸(AA)对皱纹盘鲍(初始平均体长:84.36±0.24毫米)肝胰腺中抗氧化蛋白、热休克蛋白(HSP)和核因子κB(NF-κB)转录表达模式的影响。在基础日粮中添加L-抗坏血酸-2-磷酸酯(LAMP),分别配制四种实验日粮,其AA当量分别为0.0、70.3、829.8和4967.5毫克/千克日粮。每种日粮分别投喂丙烯酸养殖槽(200升)中三组成年鲍,养殖系统为流水海水系统。每个养殖槽放养15只鲍。每天一次(17:00)投喂动物至明显饱足,持续24周。结果表明,与对照组相比,该处理组日粮AA(70.3毫克/千克)可显著上调皱纹盘鲍肝胰腺中铜/锌超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽-S-转移酶(GST)、铁蛋白(FT)和热休克蛋白26(HSP26)的表达水平。然而,锰超氧化物歧化酶、谷胱甘肽过氧化物酶(GPX)、硫氧还蛋白过氧化物酶(TPx)、硒结合蛋白(SEBP)、HSP70和HSP90的表达水平显著下调。与日粮AA为70.3毫克/千克组相比,日粮AA含量极高(4967.5毫克/千克)的皱纹盘鲍中CAT、GST和HSP26的表达水平降低。此外,与日粮AA为70.3毫克/千克组相比,日粮AA明显过量(829.8和4967.5毫克/千克)的皱纹盘鲍中锰超氧化物歧化酶、GPX、TPx、SEBP、FT、HSP70、HSP90和NF-κB的表达水平显著上调。这些结果表明,日粮AA在转录水平上影响皱纹盘鲍肝胰腺中抗氧化蛋白、热休克蛋白和NF-κB的表达水平。日粮AA含量适宜(70.3毫克/千克)通过影响抗氧化蛋白的基因表达降低了氧化应激,但日粮AA过量(829.8和4967.5毫克/千克)会诱导皱纹盘鲍产生氧化应激。