Objective: To investigate the antioxidant mechanism of diallyl sulfide (DAS) in antagonizing the reduction in peripheral blood white blood cells (WBC) induced by benzene in rats. Methods: A total of 60 specific pathogen-free adult male Sprague-Dawley rats, with a body weight of 180-220 g, were selected, and after 5 days of adaptive feeding, they were randomly divided into blank control group, DAS control group, benzene model group, benzene+low-dose DAS group, benzene+middle-dose DAS group, and benzene+high-dose DAS group, with 10 rats in each group. The rats in the benzene+low-dose DAS group, the benzene+middle-dose DAS group, the benzene+high-dose DAS group, and the DAS control group were given DAS by gavage at a dose of 40, 80, 160, and 160 mg/kg·bw, respectively, and those in the blank control group and the benzene model group were given an equal volume of corn oil; 2 hours later, the rats in the benzene model group, the benzene+low-dose DAS group, the benzene+middle-dose DAS group, and the benzene+high-dose DAS group were given a mixture of benzene (1.3 g/kg·bw) and corn oil (with a volume fraction of 50%), and those in the blank control group and the DAS control group were given an equal volume of corn oil. The above treatment was given once a day for 4 consecutive weeks. At 1 day before treatment, anticoagulated blood was collected from the jugular vein for peripheral blood cell counting. After anesthesia with intraperitoneally injected pentobarbital (50 mg/kg·bw), blood samples were collected from the abdominal aorta, serum was isolated, and the thymus, the spleen, and the femur were freed at a low temperature to measure oxidative and antioxidant indices. The femur at one side was freed for WBC counting in bone marrow. Results: Compared with the blank control group, the benzene model group had significant reductions in the volume, weight, and organ coefficient of the spleen and the thymus (P<0.05) ; compared with the benzene model group, the benzene+low-dose DAS group, the benzene+middle-dose DAS group, and the benzene+high-dose DAS group had significant increases in the volume of the spleen and the thymus and the weight and organ coefficient of the spleen (P<0.05), and the benzene+middle-dose DAS group and the benzene+high-dose DAS group had significant increases in the weight and organ coefficient of the thymus (P<0.05). Compared with the blank control group, the benzene model group had a significant reduction in WBC count in peripheral blood and bone marrow (P<0.05), and compared with the benzene model group, the benzene+middle-dose DAS group and the benzene+high-dose DAS group had a significant increase in WBC count in peripheral blood and bone marrow (P<0.05). Compared with the blank control group, the benzene model group had a significant increase in the serum level of malondialdehyde (MDA) (P<0.05) and significant reductions in total superoxide dismutase (T-SOD) activity, reduced glutathione (GSH) level, GSH/oxidized glutathione (GSSG) ratio, total antioxidant capacity (T-AOC) (P<0.05) ; compared with the benzene model group, the benzene+high-dose DAS group had a significant reduction in the serum level of MDA and significant increases in T-SOD activity, GSH level, GSH/GSSG ratio, and T-AOC (P<0.05). Compared with the blank control group, the benzene model group had a significant increase in the level of MDA (P<0.05) and significant reductions in GSH level, GSH/GSSG ratio, and T-AOC (P<0.05) in the spleen; compared with the benzene model group, the benzene+low-dose DAS group, the benzene+middle-dose DAS group, and the benzene+high-dose DAS group had a significant reduction in MDA level (P<0.05) and significant increases in GSH level and T-AOC (P<0.05), and the benzene+high-dose DAS group had significant increases in T-SOD activity and GSH/GSSG ratio (P<0.05). Compared with the blank control group, the benzene model group had a significant increase in the level of MDA in bone marrow cells (BMCs) and peripheral blood mononucleated cells (PBMCs) (P<0.05) and a significant reduction in T-AOC in PBMCs (P<0.05) ; compared with the benzene model group, the benzene+low-dose DAS group, the benzene+middle-dose DAS group, and the benzene+high-dose DAS group had a significant reduction in the level of MDA in BMCs and PBMCs (P<0.05), and the benzene+high-dose DAS group had significant increases in GSH level and GSH/GSSG ratio (P<0.05) . Conclusion: DAS can antagonize the benzene-induced reduction in peripheral blood WBC, possibly by exerting an anti-oxidative stress effect.
目的： 研究二烯丙基一硫（DAS）拮抗苯诱导大鼠外周血白细胞（WBC）降低中抗氧化作用的相关机制。 方法： SPF级成年雄性SD大鼠60只，体重180~220 g，适应性喂养5 d后，随机分为空白对照组、DAS对照组、苯模型组以及苯+DAS低、中、高剂量组，每组10只。DAS各剂量干预组和DAS对照组大鼠分别经口灌胃40、80、160、160 mg/kg·bw DAS，空白对照组和苯模型组给予等体积玉米油；2 h后，苯模型组和DAS各剂量干预组给予苯（1.3 g/kg·bw）－玉米油混合液（体积分数50%），空白对照组和DAS对照组给予等体积玉米油。1次/d，连续4周。提前1 d颈静脉取抗凝血进行外周血细胞计数；腹腔注射戊巴比妥（50 mg/kg·bw）麻醉大鼠，腹主动脉法取血并分离血清，低温剥离胸腺、脾脏和股骨，检测样品中氧化和抗氧化指标；分离一侧股骨进行骨髓中WBC计数。 结果： 与空白对照组比较，苯模型组大鼠脾脏、胸腺体积减小，重量及脏器系数均降低（P<0.05）；与苯模型组比较，DAS各剂量干预组大鼠脾脏和胸腺体积增大，脾脏重量及系数均升高（P<0.05），苯+DAS中、高剂量组胸腺重量及系数均升高（P<0.05）。与空白对照组比较，苯模型组大鼠外周血细胞和骨髓中WBC计数均降低（P<0.05）；与苯模型组比较，苯+DAS中、高剂量组外周血细胞和骨髓中WBC计数均升高（P<0.05）。与空白对照组比较，苯模型组大鼠血清中丙二醛（MDA）浓度增加（P<0.05），总超氧化物歧化酶（T-SOD）活力、还原型谷胱甘肽（GSH）含量、还原型谷胱甘肽与氧化型谷胱甘肽比值（GSH/GSSG值）以及总抗氧化能力（T-AOC）水平均降低（P<0.05）；与苯模型组比较，苯+DAS高剂量组MDA浓度降低（P<0.05），T-SOD活力、GSH含量、GSH/GSSG值以及T-AOC水平均增加（P<0.05）。与空白对照组比较，苯模型组大鼠脾脏中MDA浓度增加（P<0.05），GSH含量、GSH/GSSG值和T-AOC水平均降低（P<0.05）；与苯模型组比较，DAS各剂量干预组MDA浓度均降低（P<0.05），苯+DAS高剂量组T-SOD活力和GSH/GSSG值以及DAS各剂量干预组GSH和T-AOC水平均增加（P<0.05）。与空白对照组比较，苯模型组大鼠骨髓细胞（BMCs）和外周血单个核细胞（PBMCs）中MDA浓度均增加（P<0.05），PBMCs中T-AOC水平降低（P<0.05）；与苯模型组比较，DAS各剂量干预组BMCs和PBMCs中MDA浓度降低（P<0.05），苯+DAS高剂量组GSH含量和GSH/GSSG值均增加（P<0.05）。 结论： DAS拮抗苯诱导外周血WBC减少的机制可能与其抗氧化应激有关。.
Keywords: Benzene; Bone marrow cells; Diallyl sulfide; Leukopenia; Oxidative stress; Peripheral blood mononuclear cells; Rats.