α-硫辛酸对热应激绵羊生长性能、抗氧化及免疫的影响
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摘要
本试验旨在研究α-硫辛酸(α-LA)对热应激绵羊生长性能、抗氧化及免疫的影响,以期阐述其抗热应激的机理。选择4～5月龄、体重35～40 kg的绵羊15只,随机分为对照组(CTL)、低剂量组(LA-L,添加600 mg/kgα-LA)、高剂量组(LA-H,添加900 mg/kgα-LA),在热应激条件下(7～8月)饲喂40 d。结果表明:1)与对照组相比,α-LA添加组平均日采食量增加,低剂量组和高剂量组分别比对照组增加了7. 3%和2. 7%(P> 0. 05);高剂量组血清中丙氨酸转氨酶(ALT)、门冬氨酸转氨酶(AST)活性及α-LA添加组血浆中丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)含量显著下降(P<0.05);α-LA添加组血浆中三碘甲腺原氨酸(T3)、热休克蛋白70(HSP70)、免疫球蛋白G(IgG)含量及超氧化物歧化酶(SOD)活性显著升高(P <0.05)。2)与对照组相比,α-LA添加组肝脏中炎性相关因子IL-1β、IL-8、核因子-κB(NF-κB) mRNA表达量显著下降(P <0.05),抗炎因子干扰素-γ(IFN-γ)mRNA表达量显著升高(P<0.05)。综上,α-LA显著增强热应激绵羊的抗氧化能力,减少炎症相关因子的表达,增强抗炎因子的表达,提高机体免疫机能,且与α-LA剂量有关。
The aim of this study was to investigate the effects of α-lipoic acid( α-LA) on the growth performance,antioxidant and immunity of sheep under heat stress,in order to elucidate the mechanism of its anti-heat stress. Fifteen sheep aged 4 to 5 months and bodyweight between 35 to 40 kg were randomly selected,and equally divided into three groups: control group( CTL),low-dose group( LA-L,supplemented with 600 mg/kg α-LA) and highdose group( LA-H,supplemented with 900 mg/kg α-LA),and animals fed under heat stress( 7 to 8 months) for40 d. The results showed as follows: 1) compared with control group,average daily feed intake in α-LA group increased,and that in low-dose group and high-dose group increased by 7.3% and 2.7%( P>0.05); serum alanine aminotransferase,aspartate aminotransferase activities in high-dose group,malonaldehyde,tumor necrosis factor-α,interleukin-1β,interleukin-8 in α-LA group significantly decreased( P<0.05); the plasma contents of triiodothyronine,heat shock protein 70,immunoglobulin G,and superoxide dismutase activity in α-LA group significantly increased( P<0.05). 2) Compared with control group,the mRNA expression levels of inflammatory cytokines interleukin-1β,interleukin-8,nuclear factor-κB significantly decreased( P<0.05),and the mRNA expression levels of anti-inflammatory cytokine interferon-γ significantly increased( P<0.05). To sum up,α-LA significantly enhances the antioxidant capacity of sheep under heat stress,decreases the expression of inflammatory factors,and improves the expression of anti-inflammatory factors and immune function; the effect is related to the dosage of α-LA.
The aim of this study was to investigate the effects of α-lipoic acid( α-LA) on the growth performance,antioxidant and immunity of sheep under heat stress,in order to elucidate the mechanism of its anti-heat stress. Fifteen sheep aged 4 to 5 months and bodyweight between 35 to 40 kg were randomly selected,and equally divided into three groups: control group( CTL),low-dose group( LA-L,supplemented with 600 mg/kg α-LA) and highdose group( LA-H,supplemented with 900 mg/kg α-LA),and animals fed under heat stress( 7 to 8 months) for40 d. The results showed as follows: 1) compared with control group,average daily feed intake in α-LA group increased,and that in low-dose group and high-dose group increased by 7.3% and 2.7%( P>0.05); serum alanine aminotransferase,aspartate aminotransferase activities in high-dose group,malonaldehyde,tumor necrosis factor-α,interleukin-1β,interleukin-8 in α-LA group significantly decreased( P<0.05); the plasma contents of triiodothyronine,heat shock protein 70,immunoglobulin G,and superoxide dismutase activity in α-LA group significantly increased( P<0.05). 2) Compared with control group,the mRNA expression levels of inflammatory cytokines interleukin-1β,interleukin-8,nuclear factor-κB significantly decreased( P<0.05),and the mRNA expression levels of anti-inflammatory cytokine interferon-γ significantly increased( P<0.05). To sum up,α-LA significantly enhances the antioxidant capacity of sheep under heat stress,decreases the expression of inflammatory factors,and improves the expression of anti-inflammatory factors and immune function; the effect is related to the dosage of α-LA.
引文
[1]李富宽,杨燕,王慧,等.热应激对绵羊行为及生理的影响[J].中国农业大学学报,2018,23(2):43-49.
[2]崔艳军.热应激和氧化应激对肥育猪骨骼肌代谢的影响及硫辛酸的调控作用[D].博士学位论文.北京:中国农业科学院,2016.
[3]REED L J,DEBUSK R B G,GUNSALUS I C,et al.Crystallineα-lipoic acid:a catalytic agent associated with pyruvate dehydrogenase[J].Science,1951,114(2952):93-94.
[4]PACKER L,WITT E H,TRITSCHLER H J.alpha-Lipoic acid as a biological antioxidant[J].Free Radical and Biology Medicine,1995,19(2):227-250.
[5]尹志奎,郑斌,崔旭,等.α-硫辛酸对缺血再灌注脑神经元凋亡的作用[J].山东医药,2005,45(23):18-19.
[6]MACIAS-BARRAGAN J,CALIGIURI A,GARCA-BAN-UELOS J,et al.Effects of alpha lipoic acid and pirfenidone on liver cells antioxidant modulation against oxidative damage[J].Revista Médica de Chile,2014,142(12):1553-1564.
[7]崔珏,肖瀛,王斌,等.硫辛酸高脂日粮对小鼠免疫功能的影响[J].营养学报,2010,32(1):35-38.
[8]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.
[9]王定发,周璐丽,周汉林,等.日粮添加α-硫辛酸对海南黑山羊育肥羔羊生长性能和血液生化指标的影响[J].中国畜牧杂志,2015,51(3):33-37.
[10]吴永魁,胡仲明.动物应激医学及应激的分子调控机制[J].中国兽医学报,2005,25(5):557-560.
[11]孙长勉,栾新红,曹中赞,等.维生素E对热应激条件下肉鸡甲状腺激素分泌的影响[J].沈阳农业大学学报,2002,33(5):360-362.
[12]YANG L,TAN G Y,FU Y Q,et al.Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration,ROS production and lipid peroxidation in broiler chickens[J].Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology,2010,151(2):204-208.
[13]ZENG T,LI J J,WANG D Q,et al.Effects of heat stress on antioxidant defense system,inflammatory injury,and heat shock proteins of Muscovy and Pekin ducks:evidence for differential thermal sensitivities[J].Cell Stress and Chaperones,2014,19(6):895-901.
[14]李建文,王聪,丁斌鹰.茶多酚和硫辛酸的复合物对氧化应激肉鸭抗氧化功能的影响[J].中国饲料,2010(14):32-34.
[15]SRILATHA T,REDDY V R,QUDRATULLAH S,et al.Effect of alpha-lipoic acid and vitamin E in diet on the performance,antioxidation and immune response in broiler chicken[J].International Journal of Poultry Science,2010,9(7):678-683.
[16]王彦波,周绪霞,许梓荣.热应激蛋白70的研究进展[J].免疫学杂志,2003,19(3):S79-S82.
[17]HOROWITZ M,ROBINSON S D M.Heat shock proteins and the heat shock response during hyperthermia and its modulation by altered physiological conditions[J].Progress in Brain Research,2007,162(1):433-446.
[18]TYTELL M.Release of heat shock proteins(Hsps)and the effects of extracellular Hsps on neural cells and tissues[J].International Journal of Hyperthermia,2005,21(5):445-455.
[19]HAO Y,GU X H,WANG X L.Overexpression of heat shock protein 70 and its relationship to intestine under acute heat stress in broilers:1.Intestinal structure and digestive function[J].Poultry Science,2012,91(4):781-789.
[20]陈金雄,黄金龙,刘庆华,等.酵母铬对热应激牛免疫功能的影响[J].经济动物学报,2010,14(3):129-132.
[21]PACKER L.α-lipoic acid:a metabolic antioxidant which regulates NF-κB signal transduction and protects against oxidative injury[J].Drug Metabolism Reviews,1998,30(2):245-275.
[22]肖彧君,黄爱龙,李春霖,等.NF-κB在高浓度葡萄糖、TNF-α、IL-1β介导的血管内皮细胞损害中的作用[J].中华内分泌代谢杂志,2005,21(4):310-312.
[23]刘肖萍.多个猪Ig GⅡB类Fc受体剪接异构体的分子生物学特征[D].硕士学位论文.郑州:河南农业大学,2011.
[24]DAI Q,ZHANG C L,WU Y X,et al.CHIP activates HSF1 and confers protection against apoptosis and cellular stress[J].The EMBO Journal,2003,22(20):5446-5458.
[25]肖献忠,张华莉,刘瑛,等.HSF1及HSP70对全身炎症反应综合征的多靶点抑制作用及其机制[J].中国病理生理杂志,2010,26(10):1948-1949.
[26]岳华,黄兴,杨发龙,等.荧光定量RT-PCR检测鸡CD4、CD8基因表达水平[J].畜牧兽医学报,2008,39(6):784-790.
[2]崔艳军.热应激和氧化应激对肥育猪骨骼肌代谢的影响及硫辛酸的调控作用[D].博士学位论文.北京:中国农业科学院,2016.
[3]REED L J,DEBUSK R B G,GUNSALUS I C,et al.Crystallineα-lipoic acid:a catalytic agent associated with pyruvate dehydrogenase[J].Science,1951,114(2952):93-94.
[4]PACKER L,WITT E H,TRITSCHLER H J.alpha-Lipoic acid as a biological antioxidant[J].Free Radical and Biology Medicine,1995,19(2):227-250.
[5]尹志奎,郑斌,崔旭,等.α-硫辛酸对缺血再灌注脑神经元凋亡的作用[J].山东医药,2005,45(23):18-19.
[6]MACIAS-BARRAGAN J,CALIGIURI A,GARCA-BAN-UELOS J,et al.Effects of alpha lipoic acid and pirfenidone on liver cells antioxidant modulation against oxidative damage[J].Revista Médica de Chile,2014,142(12):1553-1564.
[7]崔珏,肖瀛,王斌,等.硫辛酸高脂日粮对小鼠免疫功能的影响[J].营养学报,2010,32(1):35-38.
[8]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.
[9]王定发,周璐丽,周汉林,等.日粮添加α-硫辛酸对海南黑山羊育肥羔羊生长性能和血液生化指标的影响[J].中国畜牧杂志,2015,51(3):33-37.
[10]吴永魁,胡仲明.动物应激医学及应激的分子调控机制[J].中国兽医学报,2005,25(5):557-560.
[11]孙长勉,栾新红,曹中赞,等.维生素E对热应激条件下肉鸡甲状腺激素分泌的影响[J].沈阳农业大学学报,2002,33(5):360-362.
[12]YANG L,TAN G Y,FU Y Q,et al.Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration,ROS production and lipid peroxidation in broiler chickens[J].Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology,2010,151(2):204-208.
[13]ZENG T,LI J J,WANG D Q,et al.Effects of heat stress on antioxidant defense system,inflammatory injury,and heat shock proteins of Muscovy and Pekin ducks:evidence for differential thermal sensitivities[J].Cell Stress and Chaperones,2014,19(6):895-901.
[14]李建文,王聪,丁斌鹰.茶多酚和硫辛酸的复合物对氧化应激肉鸭抗氧化功能的影响[J].中国饲料,2010(14):32-34.
[15]SRILATHA T,REDDY V R,QUDRATULLAH S,et al.Effect of alpha-lipoic acid and vitamin E in diet on the performance,antioxidation and immune response in broiler chicken[J].International Journal of Poultry Science,2010,9(7):678-683.
[16]王彦波,周绪霞,许梓荣.热应激蛋白70的研究进展[J].免疫学杂志,2003,19(3):S79-S82.
[17]HOROWITZ M,ROBINSON S D M.Heat shock proteins and the heat shock response during hyperthermia and its modulation by altered physiological conditions[J].Progress in Brain Research,2007,162(1):433-446.
[18]TYTELL M.Release of heat shock proteins(Hsps)and the effects of extracellular Hsps on neural cells and tissues[J].International Journal of Hyperthermia,2005,21(5):445-455.
[19]HAO Y,GU X H,WANG X L.Overexpression of heat shock protein 70 and its relationship to intestine under acute heat stress in broilers:1.Intestinal structure and digestive function[J].Poultry Science,2012,91(4):781-789.
[20]陈金雄,黄金龙,刘庆华,等.酵母铬对热应激牛免疫功能的影响[J].经济动物学报,2010,14(3):129-132.
[21]PACKER L.α-lipoic acid:a metabolic antioxidant which regulates NF-κB signal transduction and protects against oxidative injury[J].Drug Metabolism Reviews,1998,30(2):245-275.
[22]肖彧君,黄爱龙,李春霖,等.NF-κB在高浓度葡萄糖、TNF-α、IL-1β介导的血管内皮细胞损害中的作用[J].中华内分泌代谢杂志,2005,21(4):310-312.
[23]刘肖萍.多个猪Ig GⅡB类Fc受体剪接异构体的分子生物学特征[D].硕士学位论文.郑州:河南农业大学,2011.
[24]DAI Q,ZHANG C L,WU Y X,et al.CHIP activates HSF1 and confers protection against apoptosis and cellular stress[J].The EMBO Journal,2003,22(20):5446-5458.
[25]肖献忠,张华莉,刘瑛,等.HSF1及HSP70对全身炎症反应综合征的多靶点抑制作用及其机制[J].中国病理生理杂志,2010,26(10):1948-1949.
[26]岳华,黄兴,杨发龙,等.荧光定量RT-PCR检测鸡CD4、CD8基因表达水平[J].畜牧兽医学报,2008,39(6):784-790.