Department of Animal and Food Science, Texas Tech University, Lubbock, TX.
Department of Animal Biosciences, University of Guelph, Guelph, ON, Canada.
J Anim Sci. 2019 Sep 3;97(9):3871-3881. doi: 10.1093/jas/skz211.
Glutathione (GSH) is the major intracellular thiol that plays a role in numerous detoxification, bio-reduction, and conjugation reactions. The availability of Cys is thought to be the rate-limiting factor for the synthesis of GSH. The effects of immune system stimulation (ISS) on GSH levels and the GSH synthesis rate in various tissues, as well as the plasma flux of Cys, were measured in starter pigs fed a sulfur AA (SAA; Met + Cys) limiting diet. Ten feed-restricted gilts with initial body weight (BW) of 7.0 ± 0.12 kg were injected i.m. twice at 48-h intervals with either sterile saline (n = 4; ISS-) or increasing amounts of Escherichia coli lipopolysaccharide (n = 6; ISS+). The day after the second injection, pigs received a primed constant infusion of 35S-Cys (9,300 kBq/pig/h) for 5 h via a jugular catheter. Blood and tissue free Cys and reduced GSH were isolated and quantified as the monobromobimane derivatives by HPLC. The rate of GSH synthesis was determined by measurement of the specific radioactivity of GSH and tissue free Cys at the end of the infusion period. Plasma Cys and total SAA levels were reduced (16% and 21%, respectively), but plasma Cys flux was increased (26%) by ISS (P < 0.05). Immune system stimulation increased GSH levels in the plasma (48%; P < 0.05), but had no effect on GSH levels in the liver, small and large intestines, heart, muscle, spleen, kidney, lung, and erythrocytes. The fractional synthesis rate (FSR) of GSH was higher (P < 0.05) in the liver (34%), small intestine (78%), large intestine (72%), heart (129%), muscle (37%), and erythrocytes (47%) of ISS+ pigs compared to ISS- pigs. The FSR of GSH tended (P = 0.08) to be higher in the lungs (45%) of ISS+ pigs than in ISS- pigs. The absolute rate of GSH synthesis was increased by ISS (mmol/kg wet tissue/d ± SE, ISS- vs. ISS+; P < 0.05) in the liver (5.22 ± 0.22 vs. 7.20 ± 0.59), small intestine (2.54 ± 0.25 vs. 4.52 ± 0.56), large intestine (0.61 ± 0.06 vs. 1.06 ± 0.16), heart (0.21 ± 0.03 vs. 0.48 ± 0.08), lungs (1.50 ± 0.10 vs. 2.90 ± 0.21), and muscle (0.21 ± 0.03 vs. 0.34 ± 0.04), but it remained unchanged in erythrocytes, the kidney, and the spleen (P > 0.80). The current findings suggest that GSH synthesis is increased during ISS, contributing to enhanced maintenance sulfur amino acid requirements in starter pigs during ISS.
谷胱甘肽(GSH)是细胞内主要的巯基,在许多解毒、生物还原和结合反应中发挥作用。Cys 的可用性被认为是 GSH 合成的限速因素。本研究旨在测定在喂食含硫必需氨基酸(SAA;Met+Cys)受限日粮的仔猪中,免疫刺激(ISS)对各种组织中 GSH 水平和 GSH 合成率以及 Cys 血浆通量的影响。10 头初始体重(BW)为 7.0±0.12kg 的限饲母猪,每隔 48h 经肌肉注射无菌生理盐水(n=4;ISS-)或递增剂量的大肠杆菌脂多糖(n=6;ISS+)各 2 次。第二次注射后,猪通过颈内导管接受 35S-Cys(9,300kBq/猪/h)的 5h 恒速输注。通过 HPLC 以单溴双马来酰亚胺衍生物的形式分离和定量血液和组织游离 Cys 和还原型 GSH。通过在输注期末测量 GSH 和组织游离 Cys 的比放射性,确定 GSH 合成率。ISS(P<0.05)降低了血浆 Cys 和总 SAA 水平(分别为 16%和 21%),但增加了血浆 Cys 通量(26%)。ISS 增加了血浆(48%;P<0.05)中的 GSH 水平,但对肝、小肠、大肠、心脏、肌肉、脾、肾、肺和红细胞中的 GSH 水平没有影响。与 ISS-猪相比,ISS+猪的肝脏(34%)、小肠(78%)、大肠(72%)、心脏(129%)、肌肉(37%)和红细胞(47%)中的 GSH 合成率更高(P<0.05)。ISS+猪的肺(45%)中 GSH 合成率也有升高的趋势(P=0.08)。与 ISS-猪相比,ISS 增加了肝(mmol/kg 湿组织/d±SE,ISS- vs. ISS+;P<0.05)、小肠(2.54±0.25 vs. 4.52±0.56)、大肠(0.61±0.06 vs. 1.06±0.16)、心脏(0.21±0.03 vs. 0.48±0.08)、肺(1.50±0.10 vs. 2.90±0.21)和肌肉(0.21±0.03 vs. 0.34±0.04)中的 GSH 绝对合成率(P<0.05),但对红细胞、肾和脾无影响(P>0.80)。本研究表明,在 ISS 期间 GSH 合成增加,有助于满足 ISS 期间仔猪对维持硫必需氨基酸的需求。