本文關(guān)鍵詞： GnRH 主動(dòng)免疫 雄性藏綿羊 甲狀腺功能　出處：《四川農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：20世紀(jì)60年代末期,激素免疫學(xué)的產(chǎn)生為動(dòng)物生殖調(diào)控提供了新思路。用外源性GnRH主動(dòng)免疫動(dòng)物,會(huì)刺激動(dòng)物機(jī)體產(chǎn)生大量GnRH特異性抗體,引起生殖內(nèi)分泌系統(tǒng)平衡的失調(diào),改變下丘腦-垂體-性腺軸間的正常反饋調(diào)節(jié)關(guān)系,以減少生殖激素的合成和分泌及終止配子的發(fā)生,從而達(dá)到免疫去勢(shì)的目的。GnRH主動(dòng)免疫可避免外科閹割引起的動(dòng)物高度應(yīng)激、外科傷口容易感染,增加動(dòng)物發(fā)病率甚至導(dǎo)致動(dòng)物死亡等諸多不足,已成為最有望替代外科閹割的安全友好方法。本試驗(yàn)用蛋白結(jié)構(gòu)修飾后的GnRH并列體二聚物(G6k-GnRH-Tandem-Dimer)與卵清蛋白偶聯(lián),以Specol為佐劑,主動(dòng)免疫雄性藏綿羊,以研究其對(duì)雄性藏綿羊甲狀腺功能的影響。試驗(yàn)選取24周齡、體重17-19 Kg的雄性藏綿羊30只,隨機(jī)分為三組,每組10只。其中一組于25周齡時(shí)各只綿羊頸部皮下注射2 mLGnRH并列體二聚物卵清蛋白復(fù)合物(TDK-OVA)的Specol乳化劑(含TDK 100 μg),8周后加強(qiáng)免疫一次,注射劑量及方法同初次免疫,為免疫去勢(shì)組。一組在試驗(yàn)開(kāi)始前一周進(jìn)行外科手術(shù)去勢(shì),為手術(shù)去勢(shì)組。剩余一組不做任何處理,為完整對(duì)照組。初次免疫當(dāng)天記為0 wpv。初免當(dāng)天及初免后第4、8、12、16 wpv(即每4周1次,直至屠宰)頸靜脈采集血樣,酶聯(lián)免疫法(ELISA)測(cè)定血清抗體滴度及LH、FSH、T、TSH、T3、T4含量變化。加強(qiáng)免疫后8周(即16 wpv),屠宰所有試驗(yàn)綿羊,分離下丘腦、垂體及兩側(cè)睪丸。Real-Time quantitative PCR分析下丘腦、垂體生殖相關(guān)基因(GnRH-R、TSH-β、TRH、TSH-R、MT1、MT2、DIO2、DIO3) mRNA表達(dá)變化。結(jié)果顯示,免疫去勢(shì)組產(chǎn)生了良好的抗體反應(yīng),初免后機(jī)體產(chǎn)生抗體,加免后血清抗GnRH抗體滴度快速升高,由加免當(dāng)天的18.67%升高到12周(12 wpv)的最高峰(55.62%),后保持在較高水平,直至屠宰(48.85%)。與完整對(duì)照組相比,免疫去勢(shì)組血清T含量顯著下降(p0.05),屠宰時(shí)仍維持在較低水平(4.27ng/mL).與完整對(duì)照組相比,手術(shù)去勢(shì)組和免疫去勢(shì)組血清T3和T4無(wú)顯著下降(p0.05),GnRH主動(dòng)免疫顯著下調(diào)綿羊垂體GnRH-R、TSH-β及下丘腦TRH、TSH-R、DI02mRNA表達(dá)水平(p0.05),而手術(shù)去勢(shì)組綿羊垂體GnRH-R mRNA表達(dá)水平顯著高于免疫去勢(shì)組和完整對(duì)照組(p0.05)。屠宰時(shí),完整對(duì)照組睪丸平均重量為51.71±7.58 g,平均體積為49.04±6.66 cm3,免疫去勢(shì)組睪丸平均重量為41.58±5.77 g,平均體積為39.91±6.02 cm3。與完整對(duì)照組相比,免疫去勢(shì)組藏綿羊睪丸發(fā)生萎縮,其重量及體積均下降到對(duì)照組睪丸的80%(p0.05)上述研究結(jié)果表明：GnRH主動(dòng)免疫盡管使下丘腦TRH、TSH-R、DI02、DI03及垂體TSH-β mRNA表達(dá)水平下調(diào),但對(duì)血清中TSH、T3及T4濃度無(wú)明顯影響�？傊�,GnRH主動(dòng)免疫對(duì)雄性藏綿羊甲狀腺功能無(wú)不利影響。
[Abstract]:In the end of 1960s, the production of hormone immunology provided a new idea for animal reproductive regulation. Active immunization with exogenous GnRH can stimulate animal body to produce a large number of GnRH specific antibodies. It causes the imbalance of reproductive endocrine system and changes the normal feedback regulation relationship between hypothalamus-pituitary-gonad axis in order to reduce the synthesis and secretion of reproductive hormones and the termination of gametes. In order to achieve the purpose of immune castration. GnRH active immunization can avoid high stress caused by surgical castration, surgical wounds are prone to infection, increase animal morbidity and even lead to animal death and many other deficiencies. It has become the most promising safe and friendly method for surgical castration. In this study, the GnRH paratactic dimer (G6k-GnRH-Tandem-Dimer) was modified with protein structure. Coupled with ovalbumin. Male Tibetan sheep were immunized with Specol as adjuvant to study its effect on thyroid function of male Tibetan sheep. They were randomly divided into three groups. At 25 weeks of age, one sheep was subcutaneously injected with Specol emulsifier (TDK-OVA) for 2 mLGnRH paratactic dimer ovalbumin complex (TDK-OVA). TDK 100 渭 g). Once after 8 weeks, the dose and method of immunization were the same as the primary immunization group. One group underwent surgical castration one week before the trial, and the remaining group did not do any treatment. The first immunization day was 0 wpv.The blood samples were collected from jugular vein on the day of the first immunization and after the first immunization at 81216 wpv (that is once every 4 weeks until slaughtering). Elisa was used to determine the titer of serum antibody and the content of TSHT3T4 in serum. All the tested sheep were slaughtered 8 weeks after immunization. The hypothalamus, pituitary and bilateral testis. Real-time quantitative PCR were isolated to analyze the hypothalamus and pituitary gene-related gene GnRH-RnTSH- 尾. The results showed that the immune ovariectomized group had a good antibody response, and the body produced antibody after the first immunization. The titer of serum anti GnRH antibody increased rapidly from 18.67% on the day of administration to the highest peak of 55.62% at 12 weeks and remained at a higher level. Compared with the intact control group, the serum T content in the ovariectomized group decreased significantly (p 0.05). The serum T3 and T4 levels in ovariectomized group and immune castrated group were not significantly decreased compared with the control group (P 0.05). The expression of GnRH-RnTSH- 尾 and TRH-TSH-RnDI02 mRNA in hypothalamus was significantly down-regulated by GnRH (p0.05). However, the expression of GnRH-R mRNA in the pituitary gland of the ovariectomized sheep was significantly higher than that of the ovariectomized group and the intact control group. The average weight and volume of testis were 51.71 鹵7.58 g and 49.04 鹵6.66 cm ~ (3) in the intact control group and 41.58 鹵5.77 g in the castration group respectively. The average volume was 39.91 鹵6.02 cm ~ (3). Compared with the control group, the testis of Tibetan sheep in the immune castrated group were atrophied. The weight and volume decreased to 80% of the testis of the control group (p 0.05). The results showed that the hypothalamus TRH TSH-RH DI02 was induced by the active immunization of 1% GnRH. The expression of TSH- 尾 mRNA in DI03 and pituitary gland was down-regulated, but there was no significant effect on serum TSH-T _ 3 and T _ 4 levels. GnRH active immunization had no adverse effect on thyroid function of male Tibetan sheep.
【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S826

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 黃奪先,趙偉,孫益興,朱建輝,吳美珍;促性腺素釋放激素主動(dòng)免疫對(duì)公、母豬生殖機(jī)能的影響(英文)[J];江蘇農(nóng)業(yè)學(xué)報(bào);1992年02期

2 趙偉,黃奪先,孫益興,朱建輝,吳美珍,趙濟(jì)林,張金林,高建國(guó);公豬免疫去勢(shì)法初報(bào)[J];畜牧與獸醫(yī);1992年05期

3 黃奪先,趙偉,孫益興,朱建輝,吳美珍,姚連生,徐根寶;生長(zhǎng)抑素主動(dòng)免疫肉豬的增重效果[J];江蘇農(nóng)業(yè)科學(xué);1991年04期

4 周化媛;母畜對(duì)性腺類固醇抗原主動(dòng)免疫的靶器官效應(yīng)[J];國(guó)外畜牧學(xué)(草食家畜);1989年04期

5 Gibb ,雒友直;新西蘭羅姆尼母羊長(zhǎng)期主動(dòng)免疫多胎試驗(yàn)[J];國(guó)外畜牧學(xué)(草食家畜);1984年03期

6 王瑞祥,趙尚吉,蔡正華,焦淑賢,李紹楷;雄烯二酮主動(dòng)免疫提高蒙古雜交羊排卵率的研究[J];中國(guó)畜牧雜志;1988年01期

7 方富貴,章孝榮,汪存利,汪宏云,吳義師;促性腺激素釋放激素類似肽主動(dòng)免疫對(duì)公豬生殖機(jī)能的影響[J];中國(guó)畜牧雜志;2005年08期

8 黃奪先,趙偉,孫益興,朱建輝,吳美珍,姚連生,徐根寶;生長(zhǎng)抑素(Somatostatin)主動(dòng)免疫山羊的增重效果[J];江蘇農(nóng)業(yè)學(xué)報(bào);1991年04期

9 趙偉,黃奪先,朱建輝,孫益興,吳美珍;GnRH主動(dòng)免疫公豬對(duì)其性欲的影響[J];江蘇農(nóng)業(yè)科學(xué);1992年02期

10 梁皓儀;;豬病防制的基本出路:重構(gòu)動(dòng)物非特異性主動(dòng)免疫功能[J];北方牧業(yè);2009年22期

相關(guān)會(huì)議論文 前7條

1 李靜;陳代文;張克英;丁雪梅;;膽囊收縮素主動(dòng)免疫對(duì)豬內(nèi)分泌及胰腺功能的影響[A];中國(guó)畜牧獸醫(yī)學(xué)會(huì)動(dòng)物營(yíng)養(yǎng)學(xué)分會(huì)——第九屆學(xué)術(shù)研討會(huì)論文集[C];2004年

2 方富貴;章孝榮;蘇時(shí)萍;張運(yùn)海;劉亞;;GnRH主動(dòng)免疫雄大鼠胸腺內(nèi)生殖激素受體的補(bǔ)償性表達(dá)[A];中國(guó)畜牧獸醫(yī)學(xué)會(huì)動(dòng)物繁殖學(xué)分會(huì)第十六屆學(xué)術(shù)研討會(huì)論文集[C];2012年

3 蔡啟江;江朱芳;伍育英;;主動(dòng)免疫配合辨證使用中成藥治療慢性HBV感染832例[A];第十七次全國(guó)中西醫(yī)結(jié)合肝病學(xué)術(shù)會(huì)議論文匯編[C];2008年

4 蔡啟江;江朱芳;伍育英;;主動(dòng)免疫配合辨證使用中成藥治療慢性HBV感染832例[A];全國(guó)第2屆中西醫(yī)結(jié)合傳染病學(xué)術(shù)會(huì)議暨國(guó)家中醫(yī)藥管理局第1屆傳染病協(xié)作組會(huì)議論文匯編[C];2008年

5 曹鵬;;GFAP重組蛋白表達(dá)純化及其主動(dòng)免疫對(duì)脊髓損傷的神經(jīng)保護(hù)作用[A];2011中華醫(yī)學(xué)會(huì)神經(jīng)外科學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2011年

6 樸花;沈潔;郭連英;施廣霞;錢振超;;HSP70-腫瘤肽誘導(dǎo)主動(dòng)免疫保護(hù)效應(yīng)的可能機(jī)制[A];中國(guó)生理學(xué)會(huì)第21屆全國(guó)代表大會(huì)暨學(xué)術(shù)會(huì)議論文摘要匯編[C];2002年

7 韓興發(fā);曹曉涵;曾憲銀;;GnRH主動(dòng)免疫對(duì)SD大鼠下丘腦-垂體-睪丸軸生理功能的影響[A];中國(guó)畜牧獸醫(yī)學(xué)會(huì)動(dòng)物繁殖學(xué)分會(huì)第十六屆學(xué)術(shù)研討會(huì)論文集[C];2012年

相關(guān)重要報(bào)紙文章 前2條

1 吳一福;“主動(dòng)免疫”可有效治療人乳頭狀瘤病毒感染[N];中國(guó)醫(yī)藥報(bào);2002年

2 吳一福;“主動(dòng)免疫”可治療人乳頭狀瘤病毒感染[N];中國(guó)醫(yī)藥報(bào);2007年

相關(guān)博士學(xué)位論文 前5條

1 馬慧敏;基于免疫理論的非常規(guī)突發(fā)事件應(yīng)急管理主動(dòng)免疫系統(tǒng)研究[D];武漢理工大學(xué);2010年

2 袁中彪;膽囊收縮素主動(dòng)免疫對(duì)豬的營(yíng)養(yǎng)生理效應(yīng)及其機(jī)制的研究[D];四川農(nóng)業(yè)大學(xué);2004年

3 戴軍;重建乙肝主動(dòng)免疫免疫微環(huán)境的初步探討[D];四川大學(xué);2007年

4 王琴;PcrV被動(dòng)及主動(dòng)免疫在多重耐藥銅綠假單胞菌致肺損傷保護(hù)作用研究[D];復(fù)旦大學(xué);2013年

5 曹曉涵;GnRH主動(dòng)免疫動(dòng)物去勢(shì)機(jī)制及其可逆性研究[D];四川農(nóng)業(yè)大學(xué);2013年

相關(guān)碩士學(xué)位論文 前7條

1 李俊麗;三種不同蛋白構(gòu)型GnRH主動(dòng)免疫對(duì)大鼠生殖性能的影響[D];四川農(nóng)業(yè)大學(xué);2014年

2 周玉芹;GnRH主動(dòng)免疫對(duì)雄性藏綿羊甲狀腺功能的影響[D];四川農(nóng)業(yè)大學(xué);2015年

3 任曉華;GnRH主動(dòng)免疫對(duì)雄性藏綿羊免疫功能的影響[D];四川農(nóng)業(yè)大學(xué);2015年

4 李靜;膽囊收縮素主動(dòng)免疫對(duì)豬內(nèi)分泌及胰腺功能的影響[D];四川農(nóng)業(yè)大學(xué);2003年

5 趙春蕊;“主動(dòng)免疫增強(qiáng)劑”與疫苗協(xié)同對(duì)比格犬免疫功能的影響[D];四川農(nóng)業(yè)大學(xué);2012年

6 韓興發(fā);GnRH主動(dòng)免疫對(duì)大鼠下丘腦—垂體—睪丸軸及生殖反饋系統(tǒng)功能的影響[D];四川農(nóng)業(yè)大學(xué);2013年

7 李倬;豬白介素-10的表達(dá)及其主動(dòng)免疫對(duì)PRRSV滅活疫苗初次免疫效果的影響[D];南京農(nóng)業(yè)大學(xué);2010年

，

本文編號(hào)：1475817