Soybean glycinin caused NADPH-oxidase-regulated ROS overproduction and decreased ROS elimination capacity in the mid and distal intestine of juvenile grass carp (Ctenopharyngodon idella)
Soybean glycinin caused NADPH-oxidase-regulated ROS overproduction and decreased ROS elimination capacity in the mid and distal intestine of juvenile grass carp (Ctenopharyngodon idella)
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大豆甘氨酸导致幼鱼草鱼（Ctenopharyngodon idella）中段和远端肠中 NADPH 氧化酶调节的 ROS 过量产生并降低 ROS 消除能力
DOI：
10.1016/j.aquaculture.2019.734651
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发表时间：
2020-02
期刊：
影响因子：
4.5
通讯作者：
Xiao-Qiu Zhou
中科院分区：
文献类型：
--
作者：
Ya-lin Zhang;Wei-Dan Jiang;Xu-dong Duan;Lin Feng;Pei Wu;Yang Liu;Jun Jiang;Sheng-Yao Kuang;Ling Tang;Xiao-Qiu Zhou
大豆球蛋白可引起建鲤肠道活性氧（ROS）含量增加，造成肠道损伤。然而，相关的机制在很大程度上是未知的。为验证大豆球蛋白通过增加活性氧产生和降低活性氧清除能力来增加肠道活性氧含量的假说，将草鱼幼鱼分为3个处理，分别饲喂对照组、8%大豆球蛋白组和8%大豆球蛋白+1.2%谷氨酰胺组7周，测定肠道氧化损伤、活性氧含量、肠道活性氧产生和清除参数。与对照组相比，大豆球蛋白组中肠（MI）和远端肠（DI）的活性氧（ROS）、蛋白质羰基（PC）和丙二醛（MDA）含量显著升高（P< 0.05），而近端肠（PI）的含量显著降低（P≥ 0.05）。对于ROS生成参数，与对照组相比，大豆球蛋白提高了NADPH氧化酶（NOX）活性、NOX 1和NOX 2蛋白丰度以及NOX 1、NOX 2、NOX组织者（NOXO）1b、NOX激活剂（NOXA）1、p47 phox和p67 phox mRNA丰度心肌梗死后，心肌组织中NOX活性、NOX 1蛋白、NOXO 1b、NOXA 1和p22 phox mRNA丰度增加（P< 0.05）PI组除NOX 4、NOXO 1a和p47 phox基因表达量（P≥ 0.05）外，其余各基因表达量均显著低于DI组（P <0.05）。对于ROS清除参数，与对照组相比，大豆球蛋白降低了MI的总和核Nrf 2蛋白丰度、总抗氧化能力（T-AOC）、抗超氧阴离子（阿萨）、抗羟自由基（AHR）、CuZn-SOD、过氧化氢酶、谷胱甘肽（GSH）过氧化物酶（GPx）、谷胱甘肽硫转移酶（GST）和谷胱甘肽还原酶（GR）活性DI的总Nrf 2和核Nrf 2蛋白丰度、T-AOC、阿萨、AHR、CuZn-SOD、Mn-SOD、CAT、GPx和GST活性（P< 0.05）; PI的T-AOC、阿萨、CuZn-SOD、Mn-SOD和过氧化氢酶活性（P< 0.05）。与大豆球蛋白组相比，大豆球蛋白+谷氨酰胺组能显著降低草鱼心肌细胞的ROS、PC和MDA含量（P< 0.05），但对PI和DI无显著影响（P≥ 0.05）。本研究结果表明：（1）大豆球蛋白增加了MI和DI中与NOX信号通路相关的ROS产生，降低了与Nrf 2易位相关的ROS清除能力，导致ROS含量增加;（2）大豆球蛋白降低了PI中ROS产生和ROS清除能力，导致ROS含量不变;（3）谷氨酰胺对大豆球蛋白诱导的氧化应激具有保护作用，主要作用于心肌梗死。
Soybean glycinin could increase reactive oxygen species (ROS) content and cause damage in the intestine of Jian carp. However, related mechanisms are largely unknown. To test the hypothesis that glycinin increases intestinal ROS content by increasing ROS generation and decreasing ROS elimination capacity, juvenile grass carp (Ctenopharyngodon idella) were distributed into 3 treatments, fed with control, 8% glycinin or 8% glycinin+1.2% glutamine diets for 7 weeks, and intestinal oxidative damage, ROS content, intestinal ROS generation and elimination parameters were measured. Compared with control treatment, fish fed with glycinin diet had higher ROS, protein carbonyl (PC) and malondialdehyde (MDA) in the mid intestine (MI) and distal intestine (DI) (P< 0.05) rather than proximal intestine (PI) (P≥ 0.05). For ROS generation parameters, compared with control treatment, dietary glycinin increased NADPH oxidase (NOX) activities, NOX1 and NOX2 protein abundance, and NOX1, NOX2, NOX organizer (NOXO) 1b, NOX activator (NOXA) 1, p47phox and p67phox mRNA abundance (P< 0.05) of MI, increased NOX activities, NOX1 protein, NOXO1b, NOXA1 and p22phox mRNA abundance (P< 0.05) of DI, and decreased NOX activities and all the mRNA abundance of NOX family genes measured (P< 0.05) except NOX4, NOXO1a and p47phox in the PI (P≥ 0.05). For ROS elimination parameters, compared with control treatment, dietary glycinin decreased total and nuclear Nrf2 protein abundance, total antioxidant capacity (T-AOC), anti-superoxide anion (ASA), anti-hydroxy radical (AHR), CuZn-SOD, catalase, glutathione (GSH) peroxidase (GPx), GSH-S-transferase (GST) and GSH reductase (GR) activities of MI (P< 0.05), total and nuclear Nrf2 protein abundance, T-AOC, ASA, AHR, CuZn-SOD, Mn-SOD, CAT, GPx and GST activities (P< 0.05) of DI and T-AOC, ASA, CuZn-SOD, Mn-SOD and catalase activities (P< 0.05) of PI. Compared with glycinin treatment, glycinin + glutamine treatment decreased ROS, PC and MDA of MI (P< 0.05) but neither in PI nor DI of grass carp (P≥ 0.05). Concluded, this study showed that (1) glycinin increased ROS generation related to NOX signaling pathway and decreased ROS elimination capacity related to the Nrf2 translocation in the MI and DI, which lead to the increased ROS content; (2) glycinin decreased both ROS generation and ROS elimination capacity in the PI of fish, which lead to the unchanged ROS content; (3) Glutamine could protect glycinin-induced oxidative stress mainly in the MI.
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