枯草芽孢杆菌铜对先天性缺铜仔鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响
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摘要
本试验旨在研究枯草芽孢杆菌铜对先天性缺铜仔鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响。试验选择108只自然交配5 d的清洁级SD大鼠,随机分为6个组,每组18个重复,每个重复1只大鼠。Ⅰ～Ⅲ组为硫酸铜组,饲粮总铜含量分别为1.5、3.0、6.0 mg/kg;Ⅳ～Ⅵ为枯草芽孢杆菌铜组,饲粮总铜含量分别为1. 5、3. 0、6. 0 mg/kg。试验期为35 d。结果表明:1)Ⅲ组体重显著高于Ⅰ组(P<0.05),Ⅵ组体重、体长显著高于Ⅳ组(P <0.05),其他各组体重、体长、存活率差异不显著(P>0.05)。2)各组之间心脏指数、肝脏指数、脾脏指数、肾脏指数、肺脏指数差异均不显著(P>0.05)。3)Ⅱ、Ⅲ组大脑中铜沉积量显著高于Ⅰ组(P<0.05),Ⅲ组肝脏中铜沉积量显著高于Ⅰ组(P <0.05)。Ⅵ组大脑中铜沉积量显著高于Ⅴ组(P <0.05),Ⅵ组肝脏中铜沉积量显著高于Ⅳ组(P <0.05)。Ⅳ组大脑中铜沉积量显著高于Ⅰ组(P <0.05)。4)Ⅲ组血清铜蓝蛋白(CP)和碱性磷酸酶(AKP)活性显著或极显著高于Ⅰ、Ⅱ组(P <0.05或P<0.01),Ⅴ、Ⅵ组血清CP活性显著高于Ⅳ组(P<0.05),Ⅳ、Ⅴ组血清CP活性分别显著高于Ⅰ、Ⅱ组(P<0.05)。Ⅲ组血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性极显著高于Ⅰ组(P<0.01),Ⅵ组血清ALT活性显著高于Ⅳ组(P<0.05),Ⅳ组血清ALT活性显著高于Ⅰ组(P<0.05)。5)Ⅲ组血清谷胱甘肽过氧化物酶(GSH-Px)活性显著高于Ⅰ、Ⅱ组(P <0.05),Ⅲ组血清铜锌超氧化物歧化酶(CuZn-SOD)活性显著高于Ⅰ组(P<0.05);Ⅵ组血清CuZn-SOD活性显著高于Ⅳ组(P<0.05)。由此可见,适量枯草芽孢杆菌铜对先天性缺铜仔鼠生长发育和抗氧化能力具有快速修复作用,其作用效果优于硫酸铜。
This experiment was conducted to study the effects of Bacillus subtilis copper on growth and development,organ indexes,copper deposition and serum biochemical and antioxidant indexes of offspring rats with congenital copper deficiency. A total of 108 clean grade pregnant SD rats were selected and randomly divided into 6 groups with 18 replicates per group and 1 rat per replicate. Groups Ⅰ to Ⅲ were copper sulfate groups,and the dietary total copper contents were 1.5,3.0 and 6.0 mg/kg,respectively; groups Ⅳ to Ⅵ were Bacillus subtilis copper groups,and the dietary total copper contents were 1.5,3.0 and 6.0 mg/kg,respectively. The experiment lasted for 35 days. The results showed as follows: 1) the body weight of group Ⅲ was significantly higher than that of group Ⅰ (P<0.05),the body weight and body length of group Ⅵ were significantly higher than those of group Ⅳ (P<0.05),and there were no significant differences in body weight,body length and survival rate among other groups (P>0.05). 2) There were no significant differences in heart index,liver index,spleen index,kidney index and lung index among all groups (P > 0. 05). 3) The copper deposition in brain of groups Ⅱ and Ⅲ was significantly higher than that of group Ⅰ (P < 0.05),the copper deposition in liver of group Ⅲ was significantly higher than that of group Ⅰ (P < 0.05). The copper deposition in brain of group Ⅵ was significantly higher than that of group Ⅴ (P<0.05),the copper deposition in liver of group Ⅵwas significantly higher than that of group Ⅳ (P<0.05). The copper deposition in brain of group Ⅳ was significantly higher than that of group Ⅰ (P<0.05). 4) The serum ceruloplasmin (CP) and alkaline phosphatase (AKP) activities of group Ⅲ were significantly higher than those of groups Ⅰ and Ⅱ (P<0.05 or P<0.01),the serum CP activity of groups Ⅴ and Ⅵ was significantly higher than that of group Ⅳ (P < 0.05),and the serum CP activity of groups Ⅳ and Ⅴ was significantly higher than that of groups Ⅰ and Ⅱ (P < 0.05),respectively. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities of group Ⅲ were significantly higher than those of group Ⅰ (P<0.01),the serum ALT activity of group Ⅵ was significantly higher than that of group Ⅳ (P<0.05),and the serum ALT activity of group Ⅳ was significantly higher than that of group Ⅰ (P<0.05). 5) The serum glutathione peroxidase (GSH-Px) activity of group Ⅲwas significantly higher than that of groups Ⅰ and Ⅱ (P<0.05),the serum copper-zinc superoxide dismutase (CuZn-SOD) activity of group Ⅲ was significantly higher than that of group Ⅰ (P<0.05); the serum CuZnSOD activity of group Ⅵ was significantly higher than that of group Ⅳ (P<0.05). In conclusion,the appropriate amount of Bacillus Subtilis copper has a rapid repair effect on the growth and development and antioxidant capacity of offspring rats with congenital copper deficiency,and its effect is better than copper sulfate.
This experiment was conducted to study the effects of Bacillus subtilis copper on growth and development,organ indexes,copper deposition and serum biochemical and antioxidant indexes of offspring rats with congenital copper deficiency. A total of 108 clean grade pregnant SD rats were selected and randomly divided into 6 groups with 18 replicates per group and 1 rat per replicate. Groups Ⅰ to Ⅲ were copper sulfate groups,and the dietary total copper contents were 1.5,3.0 and 6.0 mg/kg,respectively; groups Ⅳ to Ⅵ were Bacillus subtilis copper groups,and the dietary total copper contents were 1.5,3.0 and 6.0 mg/kg,respectively. The experiment lasted for 35 days. The results showed as follows: 1) the body weight of group Ⅲ was significantly higher than that of group Ⅰ (P<0.05),the body weight and body length of group Ⅵ were significantly higher than those of group Ⅳ (P<0.05),and there were no significant differences in body weight,body length and survival rate among other groups (P>0.05). 2) There were no significant differences in heart index,liver index,spleen index,kidney index and lung index among all groups (P > 0. 05). 3) The copper deposition in brain of groups Ⅱ and Ⅲ was significantly higher than that of group Ⅰ (P < 0.05),the copper deposition in liver of group Ⅲ was significantly higher than that of group Ⅰ (P < 0.05). The copper deposition in brain of group Ⅵ was significantly higher than that of group Ⅴ (P<0.05),the copper deposition in liver of group Ⅵwas significantly higher than that of group Ⅳ (P<0.05). The copper deposition in brain of group Ⅳ was significantly higher than that of group Ⅰ (P<0.05). 4) The serum ceruloplasmin (CP) and alkaline phosphatase (AKP) activities of group Ⅲ were significantly higher than those of groups Ⅰ and Ⅱ (P<0.05 or P<0.01),the serum CP activity of groups Ⅴ and Ⅵ was significantly higher than that of group Ⅳ (P < 0.05),and the serum CP activity of groups Ⅳ and Ⅴ was significantly higher than that of groups Ⅰ and Ⅱ (P < 0.05),respectively. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities of group Ⅲ were significantly higher than those of group Ⅰ (P<0.01),the serum ALT activity of group Ⅵ was significantly higher than that of group Ⅳ (P<0.05),and the serum ALT activity of group Ⅳ was significantly higher than that of group Ⅰ (P<0.05). 5) The serum glutathione peroxidase (GSH-Px) activity of group Ⅲwas significantly higher than that of groups Ⅰ and Ⅱ (P<0.05),the serum copper-zinc superoxide dismutase (CuZn-SOD) activity of group Ⅲ was significantly higher than that of group Ⅰ (P<0.05); the serum CuZnSOD activity of group Ⅵ was significantly higher than that of group Ⅳ (P<0.05). In conclusion,the appropriate amount of Bacillus Subtilis copper has a rapid repair effect on the growth and development and antioxidant capacity of offspring rats with congenital copper deficiency,and its effect is better than copper sulfate.
引文
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[4]KUMAR V,KALITA J,BORA H K,et al.Temporal kinetics of organ damage in copper toxicity:a histopathological correlation in rat model[J].Regulatory Toxicology and Pharmacology,2016,81:372-380.
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[8]HUANG Y,YOO J S,KIM H J,et al.The effects of different copper(inorganic and organic)and energy(tallow and glycerol)sources on growth performance,nutrient digestibility,and fecal excretion profiles in grow ing pigs[J].Asian-Australasian Journal of Animal Sciences,2010,23(5):573-579.
[9]刘冬,黄守敏,毕璋友,等.益生菌混合培养富集锌条件优化研究[J].安徽农业科学,2015,43(20):259-262.
[10]REEVES P G,NIELSEN F H,FAHEY G C,Jr.AIN-93 purified diets for laboratory rodents:final report of the American Institute of Nutrition ad hoc w riting committee on the reformulation of the AIN-76A rodent diet[J].The Journal of Nutrition,1993,123(11):1939-1951.
[11]WANG J G,ZHU X Y,GUO Y Z,et al.Influence of dietary copper on serum grow th-related hormone levels and grow th performance of w eanling pigs[J].Biological Trace Element Research,2016,172(1):134-139.
[12]SIERPINSKA T,KONSTANTYNOWICZ J,ORYW-AL K,et al.Copper deficit as a potential pathogenic factor of reduced bone mineral density and severe tooth w ear[J].Osteoporosis International,2014,25(2):447-454.
[13]SMITH M S.Responses of chicks to dietary supplements of copper sulphate[J].British Poultry Science,1969,10(2):97-108.
[14]潘娜,朱风华,王友令,等.日粮添加高水平硫酸锌和纳米氧化铜对鸡脏器组织锌铜沉积的影响[J].中国农业科学,2011,44(23):4874-4881.
[15]O’DONOHUE J,REID M A,VARGHESE A,et al.M icronodular cirrhosis and acute liver failure due to chronic copper self-intoxication[J].European Journal of Gastroenterology&Hepatology,1993,5(7):561-562.
[16]ARAYA M,OLIVARES M,PIZARRO F,et al.Copper exposure and potential biomarkers of copper metabolism[J].Biometals,2003,16(1):199-204.
[17]CHOLEWIN'SKA E,JU S'KIEWICZ J,OGNIK K.Comparison of the effect of dietary copper nanoparticles and one copper(Ⅱ)salt on the metabolic and immune status in a rat model[J].Journal of Trace Elements in M edicine and Biology,2018,48:111-117.
[18]MEGAHED M A,HASSANIN K M A,YOUSSEF IM I,et al.Alterations in plasma lipids,glutathione and homocysteine in relation to dietary copper in rats[J].Journal of Investigational Biochemistry,2014,3(1):21-25.
[19]OZKUL H,KIRKPINAR F,MERT S,et al.Effects of high levels of dietary copper sulfate and copper proteinate on grow th performance,retention for copper and zinc of rats[J].Journal of Animal and Veterinary Advances,2011,10(11):1373-1377.
[20]ENGLE T E,SPEARS J W.Effects of dietary copper concentration and source on performance and copper status of grow ing and finishing steers[J].Journal of Animal Science,2000,78(9):2446-2451.
[21]韩丽,孔祥峰,赵越,等.枯草芽孢杆菌对围产期母猪繁殖性能和子代生长的影响[J].动物营养学报,2017,29(12):4440-4446.
[22]霍仕霞,凯赛尔·阿不都克热木,彭晓明,等.驱白巴布期对缺铜、缺锌大鼠血清中微量元素及免疫球蛋白水平的影响[J].中国现代应用药学,2011,28(8):689-694.
[23]赵春雨,张天芮,刘博,等.日粮铜来源及水平对猪血液生化指标及含铜酶活性的影响[J].中国畜牧杂志,2015,51(21):39-44.
[24]钱剑,刘国文,王哲,等.铜对软骨细胞碱性磷酸酶活性影响的实验观察[J].中国兽医杂志,2005,41(3):21-22.
[25]林立,曾晓立,张璟.丙溴磷对家兔肝组织脂质过氧化及肝功能的影响(英文)[J].中国临床康复,2004,8(21):4380-4381.
[26]张娇娇,朱风华,陈甫,等.纳米氧化铜对鸡肝细胞CP mRNA表达及培养基质中肝酶活性的影响[J].中国兽医学报,2012,32(9):1357-1361.
[27]张寿,莫重辉,常兰,等.狼毒草对小白鼠组织中的超氧化物歧化酶、谷胱甘肽过氧化物酶、丙二醛的影响[J].黑龙江畜牧兽医,2014(9):105-107.
[28]HARRIS E D.Menkes’disease:perspective and update on a fatal copper disorder[J].Nutrition Review s,1993,51(8):235-238.
[29]REZAIE A,PARKER R D,ABDOLLAHI M.Oxidative stress and pathogenesis of inflammatory bow el disease:an epiphenomenon or the cause?[J].Digestive Diseases and Sciences,2007,52(9):2015-2021.
[2]KIM B E,NEVITT T,THIELE D J.Mechanisms for copper acquisition,distribution and regulation[J].Nature Chemical Biology,2008,4(3):176-185.
[3]JU C F,YU B,ZHU D,et al.The effect of dietary sulphate copper supplementation on grow th performance,intestinal digestive enzymes and absorptive transporters in w eanling pigs[J].Journal of Animal Science and Biotechnology,2010,1(1):35-43.
[4]KUMAR V,KALITA J,BORA H K,et al.Temporal kinetics of organ damage in copper toxicity:a histopathological correlation in rat model[J].Regulatory Toxicology and Pharmacology,2016,81:372-380.
[5]SOETAN K O,OLAIYA C O,OYEWOLE O E.The importance of mineral elements for humans,domestic animals and plants:a review[J].African Journal of Food Science,2010,4(5):200-222.
[6]LEI X G,KU P K,MILLER E R,et al.Supplemental microbial phytase improves bioavailability of dietary zinc to w eanling pigs[J].The Journal of Nutrition,1993,123(6):1117-1123.
[7]ARTHINGTON J D,PATE F M,SPEARS J W.Effect of copper source and level on performance and copper status of cattle consuming molasses-based supplements[J].Journal of Animal Science,2003,81(6):1357-1362.
[8]HUANG Y,YOO J S,KIM H J,et al.The effects of different copper(inorganic and organic)and energy(tallow and glycerol)sources on growth performance,nutrient digestibility,and fecal excretion profiles in grow ing pigs[J].Asian-Australasian Journal of Animal Sciences,2010,23(5):573-579.
[9]刘冬,黄守敏,毕璋友,等.益生菌混合培养富集锌条件优化研究[J].安徽农业科学,2015,43(20):259-262.
[10]REEVES P G,NIELSEN F H,FAHEY G C,Jr.AIN-93 purified diets for laboratory rodents:final report of the American Institute of Nutrition ad hoc w riting committee on the reformulation of the AIN-76A rodent diet[J].The Journal of Nutrition,1993,123(11):1939-1951.
[11]WANG J G,ZHU X Y,GUO Y Z,et al.Influence of dietary copper on serum grow th-related hormone levels and grow th performance of w eanling pigs[J].Biological Trace Element Research,2016,172(1):134-139.
[12]SIERPINSKA T,KONSTANTYNOWICZ J,ORYW-AL K,et al.Copper deficit as a potential pathogenic factor of reduced bone mineral density and severe tooth w ear[J].Osteoporosis International,2014,25(2):447-454.
[13]SMITH M S.Responses of chicks to dietary supplements of copper sulphate[J].British Poultry Science,1969,10(2):97-108.
[14]潘娜,朱风华,王友令,等.日粮添加高水平硫酸锌和纳米氧化铜对鸡脏器组织锌铜沉积的影响[J].中国农业科学,2011,44(23):4874-4881.
[15]O’DONOHUE J,REID M A,VARGHESE A,et al.M icronodular cirrhosis and acute liver failure due to chronic copper self-intoxication[J].European Journal of Gastroenterology&Hepatology,1993,5(7):561-562.
[16]ARAYA M,OLIVARES M,PIZARRO F,et al.Copper exposure and potential biomarkers of copper metabolism[J].Biometals,2003,16(1):199-204.
[17]CHOLEWIN'SKA E,JU S'KIEWICZ J,OGNIK K.Comparison of the effect of dietary copper nanoparticles and one copper(Ⅱ)salt on the metabolic and immune status in a rat model[J].Journal of Trace Elements in M edicine and Biology,2018,48:111-117.
[18]MEGAHED M A,HASSANIN K M A,YOUSSEF IM I,et al.Alterations in plasma lipids,glutathione and homocysteine in relation to dietary copper in rats[J].Journal of Investigational Biochemistry,2014,3(1):21-25.
[19]OZKUL H,KIRKPINAR F,MERT S,et al.Effects of high levels of dietary copper sulfate and copper proteinate on grow th performance,retention for copper and zinc of rats[J].Journal of Animal and Veterinary Advances,2011,10(11):1373-1377.
[20]ENGLE T E,SPEARS J W.Effects of dietary copper concentration and source on performance and copper status of grow ing and finishing steers[J].Journal of Animal Science,2000,78(9):2446-2451.
[21]韩丽,孔祥峰,赵越,等.枯草芽孢杆菌对围产期母猪繁殖性能和子代生长的影响[J].动物营养学报,2017,29(12):4440-4446.
[22]霍仕霞,凯赛尔·阿不都克热木,彭晓明,等.驱白巴布期对缺铜、缺锌大鼠血清中微量元素及免疫球蛋白水平的影响[J].中国现代应用药学,2011,28(8):689-694.
[23]赵春雨,张天芮,刘博,等.日粮铜来源及水平对猪血液生化指标及含铜酶活性的影响[J].中国畜牧杂志,2015,51(21):39-44.
[24]钱剑,刘国文,王哲,等.铜对软骨细胞碱性磷酸酶活性影响的实验观察[J].中国兽医杂志,2005,41(3):21-22.
[25]林立,曾晓立,张璟.丙溴磷对家兔肝组织脂质过氧化及肝功能的影响(英文)[J].中国临床康复,2004,8(21):4380-4381.
[26]张娇娇,朱风华,陈甫,等.纳米氧化铜对鸡肝细胞CP mRNA表达及培养基质中肝酶活性的影响[J].中国兽医学报,2012,32(9):1357-1361.
[27]张寿,莫重辉,常兰,等.狼毒草对小白鼠组织中的超氧化物歧化酶、谷胱甘肽过氧化物酶、丙二醛的影响[J].黑龙江畜牧兽医,2014(9):105-107.
[28]HARRIS E D.Menkes’disease:perspective and update on a fatal copper disorder[J].Nutrition Review s,1993,51(8):235-238.
[29]REZAIE A,PARKER R D,ABDOLLAHI M.Oxidative stress and pathogenesis of inflammatory bow el disease:an epiphenomenon or the cause?[J].Digestive Diseases and Sciences,2007,52(9):2015-2021.