Li Shuang-An, Jiang Wei-Dan, Feng Lin, Liu Yang, Wu Pei, Jiang Jun, Kuang Sheng-Yao, Tang Ling, Tang Wu-Neng, Zhang Yong-An, Tang Xu, Shi He-Qun, Zhou Xiao-Qiu
Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China.
Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China; Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China; Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu 611130, China.
Fish Shellfish Immunol. 2017 Aug;67:475-492. doi: 10.1016/j.fsi.2017.06.032. Epub 2017 Jun 10.
In this study, we investigated the effects of dietary myo-inositol on the growth and intestinal physical barrier functions of young grass carp (Ctenopharyngodon idella). A total of 540 young grass carp (221.83 ± 0.84 g) were fed six diets containing graded levels of myo-inositol (27.0, 137.9, 286.8, 438.6, 587.7 and 737.3 mg/kg) for 10 weeks. After the growth trial, fish were challenged with Aeromonas hydrophila for 14 days. The results indicated that compared with optimal myo-inositol levels, myo-inositol deficiency (27.0 mg/kg diet): (1) decreased glutathione (GSH) contents and antioxidant enzymes activities, and down-regulated the mRNA levels of antioxidant enzymes [not glutathione-S-transferase (gst) p1 and gstp2] and NF-E2-related factor 2 (nrf2), whereas up-regulated the reactive oxygen species (ROS), malondialdehyde (MDA) and protein carbonyl (PC) contents, and the mRNA levels of Kelch-like-ECH-associated protein 1 (keap1) in three intestinal segments of young grass carp (P < 0.05). (2) Up-regulated cysteinyl aspartic acid-protease (caspase)-2, -3, -7, -8, -9, apoptotic protease activating factor-1 (apaf-1), Bcl2-associated X protein (bax), fas ligand (fasl), gen-activated protein kinase (p38mapk) and c-Jun N-terminal protein kinase (jnk) mRNA levels, whereas down-regulated B-cell lymphoma-2 (bcl-2), inhibitor of apoptosis proteins (iap) and myeloid cell leukemia-1 (mcl-1) mRNA levels in three intestinal segments of young grass carp (P < 0.05). (3) Down-regulated mRNA levels of cell cycle proteins cyclin b, cyclin d, cyclin e and E2F transcription factor 4 (e2f4) in three intestinal segments of young grass carp (P < 0.05). (4) Down-regulated the mRNA levels of zonula occludens (zo) 1, zo-2, occludin, claudin-b, -c, -f, -3c, -7a, -7b as well as -11, and up-regulated the mRNA levels of claudin-12, -15a (not -15b) and myosin light chain kinase (mlck) in three intestinal segments of young grass carp (P < 0.05). All above data indicated that dietary myo-inositol deficiency could damage physical barrier function in three intestinal segments of fish. Finally, the myo-inositol requirements based on the percent weight gain (PWG), reactive oxygen species (ROS) contents in the proximal intestine (PI), relative mRNA levels of caspase-2 (PI), cyclin b (MI) as well as claudin-b (PI) were estimated to be 276.7, 304.1, 327.9, 416.7 and 313.2 mg/kg diet, respectively.
在本研究中,我们调查了饲料中肌醇对草鱼幼鱼(Ctenopharyngodon idella)生长及肠道物理屏障功能的影响。选取540尾草鱼幼鱼(221.83 ± 0.84克),投喂含不同梯度肌醇水平(27.0、137.9、286.8、438.6、587.7和737.3毫克/千克)的六种饲料,为期10周。生长试验结束后,用嗜水气单胞菌对鱼进行攻毒14天。结果表明,与适宜肌醇水平相比,肌醇缺乏(27.0毫克/千克饲料):(1)降低了谷胱甘肽(GSH)含量和抗氧化酶活性,下调了抗氧化酶[不包括谷胱甘肽-S-转移酶(gst)p1和gstp2]及核因子E2相关因子2(nrf2)的mRNA水平,而上调了草鱼幼鱼三个肠道段的活性氧(ROS)、丙二醛(MDA)和蛋白质羰基(PC)含量,以及类Kelch样-ECH相关蛋白1(keap1)的mRNA水平(P < 0.05)。(2)上调了半胱天冬酶(caspase)-2、-3、-7、-8、-9、凋亡蛋白酶激活因子-1(apaf-1)、Bcl2相关X蛋白(bax)、Fas配体(fasl)、促分裂原活化蛋白激酶(p38mapk)和c-Jun氨基末端蛋白激酶(jnk)的mRNA水平,而下调了草鱼幼鱼三个肠道段的B细胞淋巴瘤-2(bcl-2)、凋亡抑制蛋白(iap)和髓样细胞白血病-1(mcl-1)的mRNA水平(P < 0.05)。(3)下调了草鱼幼鱼三个肠道段细胞周期蛋白cyclin b、cyclin d、cyclin e和E2F转录因子4(e2f4)的mRNA水平(P < 0.05)。(4)下调了紧密连接蛋白(zo)1、zo-2、闭合蛋白、claudin-b、-c、-f、-3c、-7a、-7b以及-11的mRNA水平,而上调了草鱼幼鱼三个肠道段claudin-12、-15a(不包括-15b)和肌球蛋白轻链激酶(mlck)的mRNA水平(P < 0.05)。上述所有数据表明,饲料中肌醇缺乏会损害鱼三个肠道段的物理屏障功能。最后,基于增重率(PWG)、近端肠道(PI)中活性氧(ROS)含量、caspase-2(PI)、cyclin b(MI)以及claudin-b(PI)的相对mRNA水平,估计草鱼幼鱼对肌醇的需求量分别为276.7、304.1、327.9、416.7和313.2毫克/千克饲料。