摘要
本试验以酿酒酵母和枯草芽孢杆菌作为混合菌种发酵小麦麸皮,通过Amberlite XAD-2柱分离纯化制备麦麸阿魏酰低聚糖(feruloyl oligosaccharides,FOs),探讨麦麸FOs对敌草快(diquat)诱导的大鼠氧化应激是否有缓解作用。试验选用体重相近的断奶雄性大鼠48只,随机分为未攻毒组、攻毒组、攻毒+100 mg/kg BW麦麸FOs组、攻毒+200 mg/kg BW麦麸FOs组、攻毒+300 mg/kg BW麦麸FOs组和攻毒+100 mg/kg BW维生素C组,每组8个重复,每个重复1只鼠,各组大鼠均饲喂相同的商业饲料。麦麸FOs和维生素C配制成水溶液,采用灌胃的方式给予,未攻毒组、攻毒组用生理盐水替代,灌胃体积0.2 mL,连续灌胃15 d。灌胃结束当天,未攻毒组大鼠注射0.3 mL生理盐水,其他5组按0.1 mmol/kg BW的剂量腹腔注射0.3 mL敌草快。敌草快攻毒12 h后取样,分析各组大鼠血浆以及肝脏、肾脏和回肠中总抗氧化能力(T-AOC),过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性以及谷胱甘肽(GSH)和8-羟基脱氧鸟苷(8-OHd G)的含量。结果显示:1)通过混菌发酵小麦麸皮制备麦麸FOs,利用Amberlite XAD-2柱进行分离纯化,获得的麦麸FOs浓度为0.059 mmol/g。2)腹腔注射敌草快显著降低大鼠血浆中SOD活性和GSH含量(P<0.05),显著降低大鼠肝脏中T-AOC,CAT、GSH-Px活性及GSH含量(P<0.05),显著降低大鼠肾脏中T-AOC及CAT、SOD活性(P<0.05),显著降低大鼠回肠中T-AOC,CAT、GSH-Px活性及GSH含量(P<0.05),并显著提高大鼠血浆和各组织中8-OHd G含量(P<0.05)。3)在敌草快引起的氧化应激状态下,灌胃一定剂量的麦麸FOs可以显著提高大鼠血浆中SOD(400 mg/kg BW)、GSH-Px活性(100和200 mg/kg BW)以及GSH含量(100和200 mg/kg BW)(P<0.05),显著提高大鼠肝脏中T-AOC(100、200和400 mg/kg BW),CAT(200和400 mg/kg BW)、SOD(100、200和400 mg/kg BW)和GSH-Px活性(100、200和400 mg/kg BW)以及GSH含量(100、200和400 mg/kg BW)(P<0.05),显著提高大鼠肾脏中T-AOC(400 mg/kg BW),CAT(200 mg/kg BW)和GSH-Px活性(200和400 mg/kg BW)以及GSH含量(400 mg/kg BW)(P<0.05),显著提高大鼠回肠中T-AOC(200 mg/kg BW),SOD(400 mg/kg BW)和GSH-Px活性(100、200和400 mg/kg BW)以及GSH含量(100、200和400 mg/kg BW)(P<0.05),显著降低血浆和各组织中8-OHd G含量(血浆、肾脏、回肠:100、200和400 mg/kg BW;肝脏:100 mg/kg BW)(P<0.05);且灌胃200、400 mg/kg BW麦麸FOs后,大鼠血浆和组织中部分抗氧化相关指标可恢复到正常生理状态水平。综上所述,本试验制备的麦麸FOs可以通过有效提高大鼠血浆和组织中抗氧酶活性和GSH含量,降低DNA氧化应激代谢产物8-OHd G的含量,有效缓解由敌草快诱导产生的氧化应激。
This study was conducted to purify wheat bran feruloyl oligosaccharides( FOs) using Amberlite XAD-2 from wheat bran fermented by Saccharomyces cerevisiae and Bacillus subtilis and subsequently investigated its relaxation effect against diquat-induced oxidative stress in rats. Forty-eight healthy weaned Wistar rats,based on the similar body weight,were randomly assigned to six groups with eight replicates in each group and one rat in each replicate. The six groups were no-challenged group,challenged group,challenged+100 mg/kg BW vitamin C group,challenged+100 mg/kg BW wheat bran FOs group,challenged+200 mg/kg BW wheat bran FOs group and challenged+400 mg/kg BW wheat bran FOs group,and rats in those group were fed the same commercial diet. Wheat bran FOs and vitamin C were made into water solutions,and the rats were orally with wheat bran FOs and vitamin C at the set doses with the perfusion volume of 0.2 mL for 15 days,while the rats in no-challenged group and challenged group were orally treated with equal volume saline with the perfusion volume of 0. 2 mL for 15 days. On the day 15,all groups,except no-challenged group,received0.1 mmol/kg BW of diquat by intraperitoneal injection at the dose of 0.3 mL. No-challenged group received an equal dose of normal saline by intraperitoneal injection. Samples were collected after challenge diquat 12 h,total antioxidative capacity( T-AOC),superoxide dismutase( SOD),glutathione peroxidase( GSH-Px),and catalase( CAT) activities,glutathione( GSH) and 8-hydroxydeoxyguanosine( 8-OHdG) contents in plasma,liver,kidney and ileum were measured. The results showed as follows: 1) The concentration of wheat branFOs was 0.059 mmol/g which fermented by mixed bacteria and purified by Amberlite XAD-2. 2) Intraperitoneal injecting diquat significantly decreased the SOD activity and GSH content in plasma( P<0.05),significantly decreased the T-AOC,CAT and GSH-Px activities and GSH content in liver( P<0.05),significantly decreased the T-AOC,CAT and SOD activities in kidney( P<0.05) and significantly decreased the T-AOC,CAT and GSH-Px activities and GSH content in ileum( P<0.05),while significantly increased the content of8-OHdG in plasma and tissues of rats( P<0.05). 3) Under the diquat-induced oxidative stress,rats oral doses of wheat bran FOs significantly improved the SOD( 400 mg/kg BW) and GSH-Px activities( 100 and200 mg/kg BW) and GSH content( 100 and 200 mg/kg BW) in plasma( P<0.05),significantly improved the T-AOC( 100,200 and 400 mg/kg BW),CAT( 200 and 400 mg/kg BW),SOD( 100,200 and400 mg/kg BW) and GSH-Px activities( 100,200 and 400 mg/kg BW) and GSH content( 100,200 and400 mg/kg BW) in liver( P < 0. 05),significantly improved the T-AOC( 400 mg/kg BW),CAT( 200 mg/kg BW) and GSH-Px activities( 200 and 400 mg/kg BW) and GSH content( 400 mg/kg BW) in kidney( P<0.05) and significantly improved the T-AOC( 200 mg/kg BW),SOD( 400 mg/kg BW) and GSH-Px activities( 100,200 and 400 mg/kg BW) and GSH content( 100,200 and 400 mg/kg BW) in ileum( P<0.05),while significantly decreased the content of 8-OHdG in plasma and tissues( plasma,kidney and ileum: 100,200 and 400 mg/kg BW; liver: 100 mg/kg BW)( P<0.05). Moreover,some antioxidant indices in plasma and tissues of rats pretreated with 200 and 400 mg/kg BW wheat bran FOs returned to normal physiological level. In summary,the wheat bran FOs prepared in this experiment exerts a protective effect against diquat-induced oxidative stress by increasing the activities of antioxidant enzyme and the content of GSH and reducing the content of 8-OHdG( DNA oxidative stress metabolite) in plasma and tissues of rats.
引文
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