麦麸阿魏酰低聚糖对敌草快致氧化应激大鼠血浆和组织抗氧化能力的影响
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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.
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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[2]ISHII T.Structure and functions of feruloylated polysaccharides[J].Plant Science,1997,127(2):111-127.
[3]葛丽花.阿魏酸低聚糖的制备及其抗氧化性质的研究[D].硕士学位论文.哈尔滨:东北林业大学,2007.
[4]曾凤彩.酶解玉米麸皮制备阿魏酰低聚糖及抗氧化性质的研究[D].硕士学位论文.齐齐哈尔:齐齐哈尔大学,2012.
[5]YAO S W,WEN X X,HUANG R Q,et al.Protection of feruloylated oligosaccharides from corn bran against oxidative stress in PC 12 Cells[J].Journal of Agricultural and Food Chem istry,2014,62(3):668-674.
[6]ZHANG H J,ZHANG S S,WANG J,et al.Wheat bran feruloyl oligosaccharides protect against AAPHinduced oxidative injury via p38M APK/PI3K-Nrf2/Keap1-M af K pathw ay[J].Journal of Functional Foods,2017,29:53-59.
[7]解春艳.茶薪菇发酵制备麦麸膳食纤维与阿魏酰低聚糖及其生物活性研究[D].博士学位论文.南京:南京农业大学,2010.
[8]陈洲.济麦22麦麸制备阿魏酰低聚糖及其生理活性研究[D].硕士学位论文.泰安:山东农业大学,2014.
[9]焦昆鹏,朱文学,马丽苹,等.玉米麸皮发酵法制备阿魏酰低聚糖及膳食纤维的研究[J].食品科技,2014(8):172-176.
[10]石璟.白藜芦醇对肥胖妊娠SD大鼠胚胎存活及后代抗氧化能力的研究[D].硕士学位论文.雅安:四川农业大学,2012.
[11]郑萍,余冰,田刚,等.精氨酸对氧化应激仔猪生长性能和血浆游离氨基酸浓度的影响[J].中国畜牧杂志,2012,48(21):38-42.
[12]郭志有.α-硫辛酸对肉鸡抗氧化的影响及其作用机制研究[D].博士学位论文.南京:南京农业大学,2014.
[13]LU T,PIAO X L,ZHANG Q,et al.Protective effects of Forsythia suspensa extract against oxidative stress induced by diquat in rats[J].Food and Chem ical Toxicology,2010,48(2):764-770.
[14]姚惠源,胡敏,袁小平,等.小麦麸皮阿魏酰低聚糖的分离与结构分析[J].食品工业科技,2009(3):79-83.
[15]SAULNIER L,VIGOUROUX J,THIBAULT J F.Isolation and partial characterization of feruloylated oligosaccharides from m aize bran[J].Carbohydrate Research,1995,272(2):241-253.
[16]袁小平.酶解麦麸制备阿魏酰低聚糖及其生物活性的研究[D].博士学位论文.无锡:江南大学,2006.
[17]王秋林,王浩毅,王树人.氧化应激状态的评价[J].中国病理生理杂志,2005,21(10):2069-2074.
[18]魏涛,高玉鹏,杜忍让,等.复合酶制剂对蛋鸡血清抗氧化酶系与肠道形态学影响[J].饲料与畜牧,2011(1):13-16.
[19]VALKO M,RHODES C J,MONCIL J,et al.Free radicals,m etals and antioxidants in oxidative stress-induced cancer[J].Chem ico-Biological Interactions,2006,160(1):1-40.
[20]张丹丹,娄鹏博,李振.GPxs家族的研究进展[J].农业技术与装备,2012(15):66-67.
[21]梅礼军,王林川.8-羟基脱氧鸟苷水平对糖尿病患者胰岛功能损害的相关性研究[J].昆明医科大学学报,2013(8):84-87.
[22]HIGUCHI M,OSHIDA J,ORINO K,et al.Wheat bran protects fischer-344 rats from diquat-induced oxidative stress by activating antioxidant system:selenium as an antioxidant[J].Bioscience,Biotechnology,and Biochem istry,2011,75(3):496-449.
[23]KATAPODIS P,VARDAKOU M,KALOGERIS E,et al.Enzym ic production of a feruloylated oligosaccharide w ith antioxidant activity from w heat flour arabinoxylan[J].European Journal of Nutrition,2003,42(1):55-60.
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