生长抑素基因疫苗pVAX1-TRX-SS对妊娠母猪及其后代的影响
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摘要
妊娠后期母猪使用电转化法肌肉注射pVAX1-TRX-SS质粒,观察其对母猪生产性能及其后代的影响。选择30头胎次相近、生理状况相似的母猪,随机分为3组,在妊娠75d时分别用活体基因导入仪注射2mg pVAX1空载体、2mg pVAX1-TRX-SS和4mg pVAX1-TRX-SS。结果表明,2 mg质粒组试验母猪后代初生重比对照组提高9.70%,4mg组比对照组高14.18%,差异都达到极显著水平(P<0.01);4mg质粒组的试验母猪后代断奶重分别比对照组和2mg质粒组提高8.16%和4.13%,差异极显著(P<0.01);检测血清中生长抑素抗体(SS-Ab)发现,4mg质粒组试验母猪在免疫后40d血清中SS-Ab的P/N极显著高于2 mg质粒组(P<0.01),同时显著高于对照组(P<0.05),阳性率达60%;对试验母猪及其后代血清激素及相关蛋白检测表明,4mg质粒组试验母猪血清催乳素(PRL)浓度比对照组高120.23%(P<0.05),两试验组母猪血清中胰岛素样生长因子1(IGF-1)有比对照组升高趋势,其中4mg组比对照组高18.54%,但差异不显著(P>0.05),试验母猪后代血清中的IGF-1未见显著差异(P>0.05);4mg质粒组试验母猪血清IGF结合蛋白(IGFBP-3)显著高于对照组和2mg质粒组(P<0.05);血清生化指标的测定结果表明,试验组母猪间及其后代间的各项指标都没有显著差异(P>0.05),故该质粒对试验母猪及其后代是安全的。本试验表明pVAX1-TRX-SS质粒能引发试验母猪产生免疫反应,提高母猪的生产性能及其后代的断奶重,且对母猪及其后代安全。
pVAX1-TRX-SS plasmids were injected into the late pregnancy sows' muscles by electric transformation,and the effects on the performance of sows and their offspring were observed.30 sows with similar fetus number and physiological condition were randomly divided into 3groups.2mg pVAX1 empty vector,2mg pVAX1-TRX-SS and 4mg pVAX1-TRX-SS was injected by electroporated with gene pulser transfection apparatus at seventy-fifth days of pregnancy.The results show that the primary weight of sow offsprings in the 2mg plasmid group was 9.70% higher than that in the control group,and the group 4 mg was 14.18% higher than the control group.The difference reached extremely significant level(P <0.01).The weaning weight of offsprings in 4mg plasmid group was 8.16%and 4.13% higher than that of the control group and the 2mg plasmid group,and the difference was extremely significant(P<0.01).Detection of somatostatin antibody(SS-Ab)in serum showed that the P/Nlevel in serum of 4mg plasmid group was significantly extremely higher than 2mg plasmid group(P<0.01)on the 40 day after immunization,while it was significantly higher than control group(P<0.05),and the positive rate reached 60%.The concentration of prolactin(PRL)of 4mg plasmid group sows was 120.23% higher than control group(P<0.05).The insulin-like growth factor 1(IGF-1)of 4mg group was 18.54% higher than control group,but the difference was not significant(P>0.05).There was no significant difference in the serum IGF-1of offspring among three groups(P>0.05).The serum IGF binding protein(IGFBP-3)of sow serum in the 4mg plasmid group was significantly higher than that of the control group and the 2mg plasmid group(P<0.05).There was no significant difference in serum biochemical indexes between the sows in each group(P>0.05),as well as their offsprings,so the plasmid was safe for the test sows and their offspring.This experiment shows that pVAX1-TRX-SS plasmid can induce immune response and improve the performance of sows.It also improves the weaning weight of the sow offspring.Importantly,it is safe for sows and their offspring.
pVAX1-TRX-SS plasmids were injected into the late pregnancy sows' muscles by electric transformation,and the effects on the performance of sows and their offspring were observed.30 sows with similar fetus number and physiological condition were randomly divided into 3groups.2mg pVAX1 empty vector,2mg pVAX1-TRX-SS and 4mg pVAX1-TRX-SS was injected by electroporated with gene pulser transfection apparatus at seventy-fifth days of pregnancy.The results show that the primary weight of sow offsprings in the 2mg plasmid group was 9.70% higher than that in the control group,and the group 4 mg was 14.18% higher than the control group.The difference reached extremely significant level(P <0.01).The weaning weight of offsprings in 4mg plasmid group was 8.16%and 4.13% higher than that of the control group and the 2mg plasmid group,and the difference was extremely significant(P<0.01).Detection of somatostatin antibody(SS-Ab)in serum showed that the P/Nlevel in serum of 4mg plasmid group was significantly extremely higher than 2mg plasmid group(P<0.01)on the 40 day after immunization,while it was significantly higher than control group(P<0.05),and the positive rate reached 60%.The concentration of prolactin(PRL)of 4mg plasmid group sows was 120.23% higher than control group(P<0.05).The insulin-like growth factor 1(IGF-1)of 4mg group was 18.54% higher than control group,but the difference was not significant(P>0.05).There was no significant difference in the serum IGF-1of offspring among three groups(P>0.05).The serum IGF binding protein(IGFBP-3)of sow serum in the 4mg plasmid group was significantly higher than that of the control group and the 2mg plasmid group(P<0.05).There was no significant difference in serum biochemical indexes between the sows in each group(P>0.05),as well as their offsprings,so the plasmid was safe for the test sows and their offspring.This experiment shows that pVAX1-TRX-SS plasmid can induce immune response and improve the performance of sows.It also improves the weaning weight of the sow offspring.Importantly,it is safe for sows and their offspring.
引文
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[13]苗树君.BST对奶牛泌乳的作用机理及其应用效果[J].黑龙江八一农垦大学学报,1999,11(3):19-23.
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[15]FLINT D J,KNIGHT C H.Interactions of prolactin and growth hormone(GH)in the regulation of mammary gland function and epithelial cell survival[J].JMammary Gland Biol Neoplasia,1997,2(1):41-48.
[16]WESTBROOK S L,ALI A M,MCDOWELL H G,et al.Passively-acquired antibodies to somatotropin release inhibiting factor(SRIF)increase appetite and growth of milk-fed lambs[J].Aust J Agric Res,1994,45(2):293-302.
[17]MCCAULEY I,BILLINGHURST A,MORGAN P,et al.Manipulation of endogenous hormones to increase growth of pigs[J].Manipulating Pig Production,1995:52-61.
[18]肖慎华.激生1号在草食动物生产中的推广应用研究[D].江苏南京:南京农业大学,2011.
[19]舒邓群,茆达干,陈荣达,等.粒细胞-巨噬细胞集落刺激因子与生长抑素融合表达质粒对断奶仔猪生长及相关激素的影响[J].南京农业大学学报,2007,30(4):92-96.
[20]LELBACH A,MUZES G,FEHER J.The insulin-like growth factor system:IGFs,IGF-binding proteins and IGFBP-proteases[J].Acta Physiol Hung,2005,92(2):97-107.
[21]姜丽红.脐血中IGF-I、IGFBP-3、GH和INS与胎儿宫内生长发育的关系[D].天津:天津医科大学,2002.
[2]顾志良,王慧娟,郁建锋.动物GH/IGF-Ⅰ轴调控机制的研究进展[J].中国畜牧兽医,2011,38(10):50-54.
[3]KHAN A S,FIOROTTO M L,CUMMINGS K K,et al.Maternal GHRH plasmid administration changes pituitary celllineage and improves progeny growth of pigs[J].Am J Physiol Endocrinol Metab,2003,285(1):224-231.
[4]SPENCER G S G,WILLIAMSON E D.Increased growth in lambs following immunization against somatostatin;preliminary observations[J].Anim Prod,1981,32:376-381.
[5]MIYAZAK J I,AIHARA H.Gene Transfer into muscle by electroporation in vivo[J].Methods Mol Med,2002,69(9):49-62.
[6]DRAGHIA-AKLI R,FIOROTTO M L,HILL L A,et al.Myogenic expression of an injectable protease-resistant growth hormone-releasing hormone augments long-term growth in pigs[J].Nat Biotechnol,1999,17(12):1179-1183.
[7]MORISHITA R,AOKI M,HASHIYA N,et al.Safety evaluation of clinical gene therapy using hepatocyte growth factor to treat peripheral arterial disease[J].Hypertension,2004,44(2):203-209.
[8]TACKET C,ROY M,WIDERA G,et al.Phase 1safety and immune response studies of a DNA vaccine encoding hepatitis B surface antigen delivered by agene delivery device[J].Vaccine,1999,17(22):2826-2829.
[9]韩燕国,梁爱心,翁士乔,等.非抗性筛选生长抑素基因疫苗对断奶仔猪促生长作用和安全性评价[C]//第五届中国畜牧科技论坛论文集,2011-09-17,重庆:中国农业出版社,2011:250-253.
[10]张德坤,杨利国,张红琳,等.抑制素基因免疫诱导单胎绵羊孪生的研究[J].中国农业大学学报,2004,9(4):40-45.
[11]徐仁宝,蒋道隆.激素对激素受体的调节[J].生理科学进展,1983(2):158-167.
[12]KHAN A S,BODLES-BRAKHOP A M,FIOROT-TO M L,et al.Effects of maternal plasmid GHRHtreatment on offspring growth[J].Vaccine,2010,28(8):1905-1910.
[13]苗树君.BST对奶牛泌乳的作用机理及其应用效果[J].黑龙江八一农垦大学学报,1999,11(3):19-23.
[14]SVENNERSTEN-SJAUNJA K,OLSSON K.Endocrinology of milk production[J].Domest Anim Endocrinol,2005,29(2):241-258.
[15]FLINT D J,KNIGHT C H.Interactions of prolactin and growth hormone(GH)in the regulation of mammary gland function and epithelial cell survival[J].JMammary Gland Biol Neoplasia,1997,2(1):41-48.
[16]WESTBROOK S L,ALI A M,MCDOWELL H G,et al.Passively-acquired antibodies to somatotropin release inhibiting factor(SRIF)increase appetite and growth of milk-fed lambs[J].Aust J Agric Res,1994,45(2):293-302.
[17]MCCAULEY I,BILLINGHURST A,MORGAN P,et al.Manipulation of endogenous hormones to increase growth of pigs[J].Manipulating Pig Production,1995:52-61.
[18]肖慎华.激生1号在草食动物生产中的推广应用研究[D].江苏南京:南京农业大学,2011.
[19]舒邓群,茆达干,陈荣达,等.粒细胞-巨噬细胞集落刺激因子与生长抑素融合表达质粒对断奶仔猪生长及相关激素的影响[J].南京农业大学学报,2007,30(4):92-96.
[20]LELBACH A,MUZES G,FEHER J.The insulin-like growth factor system:IGFs,IGF-binding proteins and IGFBP-proteases[J].Acta Physiol Hung,2005,92(2):97-107.
[21]姜丽红.脐血中IGF-I、IGFBP-3、GH和INS与胎儿宫内生长发育的关系[D].天津:天津医科大学,2002.