摘要
目的:探讨GnRH激动剂(GnRHa)主动免疫对绵羊生殖激素合成与分泌的作用,并深入研究GnRH-A免疫调节动物生殖功能的机理。方法:42只5～6月龄母绵羊(Ovis aries)随机分为6组(n=7),EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别于0和14天皮下注射阿拉瑞林抗原200、300和400μg;EG-Ⅳ和EG-Ⅴ分别皮下注射阿拉瑞林抗原200、300、0、7、14和21天各一次,共4次;对照组在0和14天皮下注射抗原溶媒(除不用阿拉瑞林外,其余成分和制备方法与阿拉瑞林抗原相同)2.0 ml。无菌采集不同时段的血液,分离血清。以ELISA测定血清GnRH抗体浓度,用激素检测试剂盒(ELISA)分别测定血清GnRH、FSH、LH和E2浓度。结果:①阿拉瑞林首次免疫7天后,各实验组的抗体浓度逐渐升高,EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别在28、28和35天达到峰值(P<0.05),EG-IV和EG-V则在在45天达到峰值(P<0.01),至60天时仍明显高于对照组(P<0.05)。14～60天间EG-IV和EG-V抗体浓度均高于EG-Ⅰ、EG-Ⅱ和EG-Ⅲ(P<0.05)。②EG-Ⅰ、EG-Ⅱ的GnRH在21和28天抵谷值(P<0.05),EG-Ⅲ、EG-Ⅳ和EG-Ⅴ则在45天抵谷值(P<0.01),且以EG-Ⅴ为最低。谷值之后逐渐上升趋势,70天时达到免疫注射前水平。③实验组血清FSH浓度始终高于对照组(P<0.05)。EG-Ⅰ、EG-Ⅱ和EG-Ⅲ于28、28和35天达到峰值(P<0.05),而EG-IV和EG-V在60天达到高峰值(P<0.01)。④实验组绵羊血清LH呈下降趋势,EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别在21、21和28天达到谷值(P<0.01),EG-Ⅳ和EG-Ⅴ在35天达到谷值(P<0.01)。35天时EG-Ⅳ和EG-Ⅴ低于EG-Ⅰ、EG-Ⅱ和EG-Ⅲ。⑤各组的血清E2含量无显著差异。结论:GnRH激动剂(阿拉瑞林)抗原主动免疫可促进GnRH抗体的生成,抑制母羊GnRH和LH的合成与分泌,增强FSH的合成与分泌,且随着注射剂量和注射次数的增加,这种作用更加明显,而对血清E2无明显影响。
Objective：To explore the characteristics of reproductive hormones synthesis and secretion in ewes actively immunized with GnRH agonist,further to study the mechanisms of GnRH agonists regulating reproductive functions in animals.Methods：Forty-two ewes were randomly assigned into six groups（each of 7 eves）.Ewes in experiment group（EG）-Ⅰ,EG-Ⅱ and EG-Ⅲ were injected subcutaneously twice（at days 0 and 7） with 200,300 and 400 μg GnRH agonist（Alarelin） antigen respectively.Animals in EG-Ⅳ and EG-Ⅴwere injected subcutaneously with 200 μg and 300 μg Alarelin antigen for four times（at days 0,7,14 and 21） respectively.The control group（CG） was injected with 2.0 ml solvent subcutaneously twice（at days 0 and 7）.Blood samples were collected from jugular vein.Serum concentrations of anti-GnRH antibody,GnRH,FSH,LH and estradiol（E2） were detected using ELISA.Results：（1） Anti-GnRH antibody was detected at days 7 after the first injection of alarelin antigen,antibody concentrations in EG-Ⅰ,EG-Ⅱ and EG-Ⅲ reached peak levels at days 28,28 and 35 respectively.Both EG-Ⅳ and EG-Ⅴ reached the peak values at days 45.Serum concentrations of GnRH antibody in five experiment groups were higher than that CG（P0.05） from days 14 to 70.（2） When compared to CG,serum GnRH concentrations in EG-Ⅰ,EG-Ⅱ,EG-Ⅲ,EG-Ⅳ and EG-Ⅴreached the bottom values at days 21,28（P0.05）,45,45 and 45（P0.01）.GnRH concentrations in EG-Ⅴ was the lowest.（3） Serum FSH concentrations in experiment groups began to increase gradually from days 14.FSH concentrations in experiment groups were higher than that in CG during whole experiment.（4）Serum LH concentrations in experiment groups declined compared to CG.At days 35,EG-Ⅳ and EG-Ⅴwere lower than EG-Ⅰ,EG-Ⅱ and EG-Ⅲ.（5） Serum estradiol levels had no significant differences among all groups.Conclusion：GnRH agonist（alarelin）antigen active immunization could stimulate production of GnRH antibody,promote secretion
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第2期132-136,共5页
Chinese Journal of Immunology
基金
国家自然科学基金项目(No.31060350)
