谷氨酰胺对脂多糖诱导的断奶仔猪氧化应激的影响
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摘要
本试验以大肠杆菌型脂多糖(LPS)建立氧化应激模型,探讨了谷氨酰胺(GLN)对断奶仔猪氧化应激的影响。选用24头28日龄的健康三元(杜×长×大)断奶仔猪,随机分成3组,每组8个重复,每个重复1头猪。对照组和应激组饲喂基础饲粮,GLN组饲粮在基础饲粮中添加1%GLN,试验期为30 d。在试验第22、25、28和30天,应激组和GLN组仔猪分别按每千克体重腹腔注射100μg LPS,对照组仔猪腹腔注射相同剂量的灭菌生理盐水,第30天进行前腔静脉采血并屠宰采取所需肠道样品,检测氧化应激相关指标。结果显示:1)LPS攻毒前各组血清抗氧化能力指标均无显著差异(P>0.05)。LPS攻毒后,应激组血清丙二醛(MDA)含量显著高于对照组(P<0.05);GLN组血清MDA含量和超氧化物歧化酶(SOD)活性显著低于应激组和对照组(P<0.05)。2)LPS攻毒后,在十二指肠黏膜中,GLN组过氧化氢酶(CAT)和锌铜超氧化物歧化酶(Cu Zu SOD)基因相对表达量显著高于应激组(P<0.05)。在空肠黏膜中,GLN组CAT、锰超氧化物歧化酶(Mn SOD)、谷胱甘肽过氧化物酶1(GPX1)和谷胱甘肽过氧化物酶4(GPX4)基因相对表达量显著高于对照组和应激组(P<0.05),对照组GPX4基因相对表达量显著高于应激组(P<0.05)。在回肠黏膜中,GLN组和应激组CAT基因相对表达量显著低于对照组(P<0.05),GPX4基因相对表达量显著高于对照组(P<0.05);GLN组Mn SOD基因相对表达量显著高于对照组和应激组(P<0.05),Cu Zn SOD基因相对表达量显著高于对照组(P<0.05)。结果表明,GLN在一定程度上可以缓解断奶仔猪因LPS引起的氧化应激,以期为实际生产中减少氧化应激提供一定的理论基础。
This study was conducted to evaluate the effects of glutamine(GLN)on oxidative stress of weaned piglets based on the oxidative stress model by Escherichia coli lipopolysaccharide(LPS).Twenty-four 28-dayold healthy crossbred(Duroc×Landrace×Largewhite)weaned piglets were randomly assigned into three groups with eight replicates per group and one piglet per replicate.The piglets in control group and stress group were fed the basal diets,and the others in GLN group were fed the basal diet supplemented with 1%GLN.The experiment lasted for 30 days.At days 22,25,28 and 30 of the experiment,the piglets in stress group and GLN group were injected intraperitoneally with 100μg LPS per kg body weight,whereas piglets in control group were injected intraperitoneally with the same dosage of sterile saline.At day 30,blood samples were collected by precaval vein and the intestinal tract samples were collected after slaughter for detection of the oxidative stress related indexes.The results showed as follows:1)serum antioxidant capacity indexes had no significant differences among all groups before LPS challenge(P>0.05).After LPS challenge,the content of malondialdehyde(MDA)in serum in stress group was significantly higher than that in control group(P<0.05),and the content of MDA and the activity of superoxide dismutase(SOD)in serum in GLN group were significantly lower than those in stress group and control group(P<0.05).2)After LPS challenge,the relative gene expression levels of catalase(CAT)and Cu-Zn superoxide dismutase(Cu Zu SOD)in duodenum mucosa in GLN group were significantly higher than those in stress group(P<0.05).The relative gene expression levels of CAT,manganese superoxide dismutase(Mn SOD),glutathione peroxidase 1(G PX1)and glutathione peroxidase 4(G PX4)in jejunum mucosa in GLN group were significantly higher than those in control group and stress group(P<0.05),and the relative gene expression levels of G PX4 in control group was significantly higher than that in stress group(P<0.05).The relative gene expression level of CAT in ileum mucosa in GLN group and stress group was significantly lower than that in control group(P<0.05),and the relative gene expression level of G PX4 was significantly higher than that in control group(P<0.05).The relative gene expression level of Mn SOD in ileum mucosa in GLN group was significantly higher than that in control group and stress group(P<0.05),and the relative gene expression level of Cu Zn SOD was significantly higher than that in control group(P<0.05).The results suggest that GLN can alleviate oxidative stress of weaned piglets challenged by LPS to some extent,thereby provide a theoretical basis for reducing oxidative stress in actual production.
This study was conducted to evaluate the effects of glutamine(GLN)on oxidative stress of weaned piglets based on the oxidative stress model by Escherichia coli lipopolysaccharide(LPS).Twenty-four 28-dayold healthy crossbred(Duroc×Landrace×Largewhite)weaned piglets were randomly assigned into three groups with eight replicates per group and one piglet per replicate.The piglets in control group and stress group were fed the basal diets,and the others in GLN group were fed the basal diet supplemented with 1%GLN.The experiment lasted for 30 days.At days 22,25,28 and 30 of the experiment,the piglets in stress group and GLN group were injected intraperitoneally with 100μg LPS per kg body weight,whereas piglets in control group were injected intraperitoneally with the same dosage of sterile saline.At day 30,blood samples were collected by precaval vein and the intestinal tract samples were collected after slaughter for detection of the oxidative stress related indexes.The results showed as follows:1)serum antioxidant capacity indexes had no significant differences among all groups before LPS challenge(P>0.05).After LPS challenge,the content of malondialdehyde(MDA)in serum in stress group was significantly higher than that in control group(P<0.05),and the content of MDA and the activity of superoxide dismutase(SOD)in serum in GLN group were significantly lower than those in stress group and control group(P<0.05).2)After LPS challenge,the relative gene expression levels of catalase(CAT)and Cu-Zn superoxide dismutase(Cu Zu SOD)in duodenum mucosa in GLN group were significantly higher than those in stress group(P<0.05).The relative gene expression levels of CAT,manganese superoxide dismutase(Mn SOD),glutathione peroxidase 1(G PX1)and glutathione peroxidase 4(G PX4)in jejunum mucosa in GLN group were significantly higher than those in control group and stress group(P<0.05),and the relative gene expression levels of G PX4 in control group was significantly higher than that in stress group(P<0.05).The relative gene expression level of CAT in ileum mucosa in GLN group and stress group was significantly lower than that in control group(P<0.05),and the relative gene expression level of G PX4 was significantly higher than that in control group(P<0.05).The relative gene expression level of Mn SOD in ileum mucosa in GLN group was significantly higher than that in control group and stress group(P<0.05),and the relative gene expression level of Cu Zn SOD was significantly higher than that in control group(P<0.05).The results suggest that GLN can alleviate oxidative stress of weaned piglets challenged by LPS to some extent,thereby provide a theoretical basis for reducing oxidative stress in actual production.
引文
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[2]于国江,冯会中,郑鹏飞,等.仔猪运输综合征诊治[J].中国猪业,2009,4(9):43-44.
[3]王蕾,刘坚,侯永清,等.α-酮戊二酸对LPS慢性应激仔猪小肠黏膜形态与功能的影响[J].畜牧兽医学报,2010,41(1):46-52.
[4]刘坚,侯永清,丁斌鹰,等.α-酮戊二酸对脂多糖应激断奶仔猪空肠黏膜蛋白合成和抗氧化能力的影响[J].中国畜牧杂志,2010,46(11):35-38.
[5]林海.家禽热应激状态下的营养与生理反应[M]//李德发.动物营养研究进展.北京:中国农业科学技术出版社,2004:237-248.
[6]CURI R,LAGRANHA C J,DOI S Q,et al.Molecular mechanisms of glutamine action[J].Journal of Cellular Physiology,2005,204(2):392-401.
[7]黄冠庆,林红英.谷氨酰胺对高温环境下黄羽肉鸡抗氧化能力的影响[J].中国饲料,2006(20):24-26.
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[12]杨彩梅,徐卫丹,陈安国.甘氨酰-L-谷氨酰胺对断奶仔猪生长性能及肠道吸收功能的影响[J].中国畜牧杂志,2005,41(8):6-8.
[13]ZHANG H J,GUO Y M,TIAN Y D,et al.Dietary conjugated linoleic acid improves antioxidant capacity in broiler chicks[J].British Poultry Science,2008,49(2):213-221.
[14]GHOSH B,HANEVOLD C D,DOBASHI K,et al.Tissue differences in antioxidant enzyme gene expression in response to endotoxin[J].Free Radical Biology and M edicine,1996,21(4):533-540.
[15]ABD-ALLAH A R A,HELAL G K,AL-YAHYA A A,et al.Pro-inflammatory and oxidative stress pathw ays w hich compromise sperm motility and survival may be altered by L-carnitine[J].Oxidative M edicine and Cellular Longevity,2009,2(2):73-81.
[16]DENNO R,ROUNDS J D,FARIS R,et al.Glutamineenriched total parenteral nutrition enhances plasma glutathione in the resting state[J].Journal of Surgical Research,1996,61(1):35-38.
[17]边连全,陈静,刘显军.谷氨酰胺对早期断奶仔猪小肠粘膜形态和抗氧化性能的影响[J].沈阳农业大学学报,2006,37(6):849-852.
[18]郑荣梁,黄中洋.自由基生物学[M].3版.北京:高等教育出版社,2007.
[19]ROTRUCK J T,POPE A L,GANTHER H E,et al.Selenium:biochemical role as a component of glutathione peroxidase[J].Science,1973,179(4073):588-590.
[20]王晓雅.硒蛋白的抗氧化作用[J].现代农业科技,2006(8):183-184.
[21]SUN Q,KOJIMA H,KOMURA S,et al.Effect of selenium on human phospholipid hydroperoxide glutathione peroxidase expression and host cell susceptibility to lipid hydroperoxide-mediated injury[J].Biochemistry and M olecular Biology International,1997,42(5):957-963.
[22]NOOR R,MITTAL S,IQBAL J.Superoxide dismutaseapplications and relevance to human diseases[J].M edical Science M onitor,2002,8(9):210-215.
[2]于国江,冯会中,郑鹏飞,等.仔猪运输综合征诊治[J].中国猪业,2009,4(9):43-44.
[3]王蕾,刘坚,侯永清,等.α-酮戊二酸对LPS慢性应激仔猪小肠黏膜形态与功能的影响[J].畜牧兽医学报,2010,41(1):46-52.
[4]刘坚,侯永清,丁斌鹰,等.α-酮戊二酸对脂多糖应激断奶仔猪空肠黏膜蛋白合成和抗氧化能力的影响[J].中国畜牧杂志,2010,46(11):35-38.
[5]林海.家禽热应激状态下的营养与生理反应[M]//李德发.动物营养研究进展.北京:中国农业科学技术出版社,2004:237-248.
[6]CURI R,LAGRANHA C J,DOI S Q,et al.Molecular mechanisms of glutamine action[J].Journal of Cellular Physiology,2005,204(2):392-401.
[7]黄冠庆,林红英.谷氨酰胺对高温环境下黄羽肉鸡抗氧化能力的影响[J].中国饲料,2006(20):24-26.
[8]DAHLQVIST A.Method for assay of intestinal disaccharidases[J].Analytical Biochemistry,1964,7(1):18-25.
[9]SCHMITTGEN T D,LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocols,2008,3(6):1101-1108.
[10]陈瑾,周小秋,冯琳.谷氨酰胺对动物肠道抗氧化能力的影响[J].饲料工业,2009,30(2):55-57.
[11]曹婧然,谢颖,李辉,等.谷氨酰胺在氧化应激疾病中的作用及其机制的研究[J].临床误诊误治,2013,26(9):102-104.
[12]杨彩梅,徐卫丹,陈安国.甘氨酰-L-谷氨酰胺对断奶仔猪生长性能及肠道吸收功能的影响[J].中国畜牧杂志,2005,41(8):6-8.
[13]ZHANG H J,GUO Y M,TIAN Y D,et al.Dietary conjugated linoleic acid improves antioxidant capacity in broiler chicks[J].British Poultry Science,2008,49(2):213-221.
[14]GHOSH B,HANEVOLD C D,DOBASHI K,et al.Tissue differences in antioxidant enzyme gene expression in response to endotoxin[J].Free Radical Biology and M edicine,1996,21(4):533-540.
[15]ABD-ALLAH A R A,HELAL G K,AL-YAHYA A A,et al.Pro-inflammatory and oxidative stress pathw ays w hich compromise sperm motility and survival may be altered by L-carnitine[J].Oxidative M edicine and Cellular Longevity,2009,2(2):73-81.
[16]DENNO R,ROUNDS J D,FARIS R,et al.Glutamineenriched total parenteral nutrition enhances plasma glutathione in the resting state[J].Journal of Surgical Research,1996,61(1):35-38.
[17]边连全,陈静,刘显军.谷氨酰胺对早期断奶仔猪小肠粘膜形态和抗氧化性能的影响[J].沈阳农业大学学报,2006,37(6):849-852.
[18]郑荣梁,黄中洋.自由基生物学[M].3版.北京:高等教育出版社,2007.
[19]ROTRUCK J T,POPE A L,GANTHER H E,et al.Selenium:biochemical role as a component of glutathione peroxidase[J].Science,1973,179(4073):588-590.
[20]王晓雅.硒蛋白的抗氧化作用[J].现代农业科技,2006(8):183-184.
[21]SUN Q,KOJIMA H,KOMURA S,et al.Effect of selenium on human phospholipid hydroperoxide glutathione peroxidase expression and host cell susceptibility to lipid hydroperoxide-mediated injury[J].Biochemistry and M olecular Biology International,1997,42(5):957-963.
[22]NOOR R,MITTAL S,IQBAL J.Superoxide dismutaseapplications and relevance to human diseases[J].M edical Science M onitor,2002,8(9):210-215.