Wuxi Fisheries College, , Wuxi 214081, China.
Wuxi Fisheries College, , Wuxi 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Fish Shellfish Immunol. 2018 Jul;78:69-78. doi: 10.1016/j.fsi.2018.04.028. Epub 2018 Apr 18.
The present study assessed the effects of dietary arginine on antioxidant status and immunity involved in AMPK-NO signaling pathway in juvenile blunt snout bream. Fish were fed six practical diets with graded arginine levels ranging from 0.87% to 2.70% for 8 weeks. The results showed that compared with the control group (0.87% dietary arginine level), significantly higher mRNA levels of adenosine monophosphate activated protein kinase (AMPK) and nitric oxide synthetase (NOS), activities of total nitric oxide synthetase (T-NOS) and nitric oxide synthetase (iNOS), and plasma nitric oxide (NO) contents were observed in fish fed with 1.62%-2.70% dietary arginine levels. Significantly higher levels of NOS and iNOS were observed in fish fed with 1.62%-2.70% dietary arginine levels in enzyme-linked immune sorbent assay. At dietary arginine levels of 1.22%-2.70%, the mRNA levels of iNOS were significantly improved. Dietary arginine also significantly influenced plasma interleukin 8 (IL-8) and tumour necrosis factor-α (TNF-α) contents. Furthermore, dietary arginine significantly affected the activity and mRNA level of glutathione peroxidase (GPx), the mRNA levels of pro-inflammatory factor including IL-8 and TNF-α and plasma malondialdehyde (MDA) content. However, total superoxide dismutase (T-SOD) activity, plasma complement component 3 (C3) content, plasma immunoglobulin M (IgM) content, plasma interleukin 1β (IL-1β) content and the mRNA levels of copperzinc superoxide dismutase (Cu/Zn-SOD), manganese superoxide dismutase (Mn-SOD) and IL-1β were not significantly affected by dietary arginine. After Aeromonas hydrophila challenge, the death rate was significantly lowered in fish fed with 1.62%-1.96% dietary arginine levels. Furthermore, the mRNA levels of AMPK, NOS and iNOS, plasma NO content and the activities of T-NOS and iNOS showed an upward trend with increasing dietary arginine levels. Significantly higher levels of NOS and iNOS were observed in fish fed with 1.62%-2.70% dietary arginine levels in enzyme-linked immune sorbent assay. At dietary arginine levels of 1.96%-2.31%, T-SOD activities were significantly improved. Significantly higher GPx activities were observed in fish fed with 1.22%-2.70% dietary arginine levels. At dietary arginine levels of 1.22%-2.31%, the plasma TNF-α and IL-8 contents were significantly decreased. Significantly lower plasma IL-1β contents were observed in fish fed 1.62%-1.96% dietary arginine levels. Dietary arginine significantly influenced the mRNA levels of antioxidant and pro-inflammatory genes including Cu/Zn-SOD, Mn-SOD, GPx, IL-8, TNF-α and IL-1β. Significantly higher plasma C3 contents and significantly lower plasma MDA contents were observed in fish fed with 1.62%-1.96% arginine levels. Furthermore, plasma IgM contents were significantly improved at dietary arginine levels of 1.62%-2.31%. However, high dietary arginine group (2.70%) significantly improved the mRNA levels of pro-inflammatory genes including IL-8, TNF-α and IL-1β and plasma MDA, IL-8, TNF-α and IL-1β contents as compared with optimal dietary arginine levels (1.62% and 1.96%). The present results indicate that optimal arginine level (1.62% and 1.96%) could improve antioxidant capacity, immune response and weaken tissues inflammatory involved in arginine-AMPK-NO signaling pathway, while high arginine level resulted in excessive NO production, leading to increase oxidative stress damage and inflammatory response in juvenile blunt snout bream.
本研究评估了饲料精氨酸对参与 AMPK-NO 信号通路的幼龄钝吻鲷抗氧化状态和免疫的影响。鱼被喂食六种具有从 0.87%到 2.70%不等的精氨酸水平的实用饲料 8 周。结果表明,与对照组(0.87%的饲料精氨酸水平)相比,1.62%-2.70%的饲料精氨酸水平组鱼的腺苷单磷酸激活蛋白激酶(AMPK)和一氧化氮合酶(NOS)的 mRNA 水平、总一氧化氮合酶(T-NOS)和一氧化氮合酶(iNOS)的活性以及血浆一氧化氮(NO)含量显著升高。酶联免疫吸附试验中,1.62%-2.70%的饲料精氨酸水平组鱼的 NOS 和 iNOS 水平显著升高。在 1.22%-2.70%的饲料精氨酸水平下,iNOS 的 mRNA 水平显著提高。饲料精氨酸还显著影响血浆白细胞介素 8(IL-8)和肿瘤坏死因子-α(TNF-α)含量。此外,饲料精氨酸显著影响谷胱甘肽过氧化物酶(GPx)的活性和 mRNA 水平、包括 IL-8 和 TNF-α在内的促炎因子的 mRNA 水平以及血浆丙二醛(MDA)含量。然而,总超氧化物歧化酶(T-SOD)活性、血浆补体成分 3(C3)含量、血浆免疫球蛋白 M(IgM)含量、血浆白细胞介素 1β(IL-1β)含量以及铜锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)和 IL-1β的 mRNA 水平不受饲料精氨酸的影响。在遭受嗜水气单胞菌感染后,1.62%-1.96%饲料精氨酸水平组的死亡率显著降低。此外,随着饲料精氨酸水平的增加,AMPK、NOS 和 iNOS 的 mRNA 水平、血浆 NO 含量以及 T-NOS 和 iNOS 的活性呈上升趋势。酶联免疫吸附试验中,1.62%-2.70%的饲料精氨酸水平组鱼的 NOS 和 iNOS 水平显著升高。在 1.96%-2.31%的饲料精氨酸水平下,T-SOD 活性显著提高。1.22%-2.70%的饲料精氨酸水平组鱼的 GPx 活性显著提高。在 1.22%-2.31%的饲料精氨酸水平下,血浆 TNF-α和 IL-8 含量显著降低。1.62%-1.96%饲料精氨酸水平组鱼的血浆 IL-1β含量显著降低。饲料精氨酸显著影响抗氧化和促炎基因如 Cu/Zn-SOD、Mn-SOD、GPx、IL-8、TNF-α和 IL-1β的 mRNA 水平。1.62%-1.96%的饲料精氨酸水平组鱼的血浆 C3 含量显著升高,血浆 MDA 含量显著降低。此外,在 1.62%-2.31%的饲料精氨酸水平下,血浆 IgM 含量显著提高。然而,高饲料精氨酸组(2.70%)与最佳饲料精氨酸水平(1.62%和 1.96%)相比,显著提高了促炎基因如 IL-8、TNF-α和 IL-1β以及血浆 MDA、IL-8、TNF-α和 IL-1β的含量。结果表明,最佳精氨酸水平(1.62%和 1.96%)可以提高抗氧化能力、免疫反应并减弱参与精氨酸-AMPK-NO 信号通路的组织炎症,而高精氨酸水平会导致过多的 NO 产生,从而导致幼龄钝吻鲷的氧化应激损伤和炎症反应增加。