华中农业大学学报
華中農業大學學報
화중농업대학학보
JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY
2013年
4期
77-83
,共7页
魏锁成%巩转娣%安利峰%谢坤%李玉孔
魏鎖成%鞏轉娣%安利峰%謝坤%李玉孔
위쇄성%공전제%안리봉%사곤%리옥공
阿拉瑞林%FSH受体%蛋白表达%组织学结构%子宫%母羊
阿拉瑞林%FSH受體%蛋白錶達%組織學結構%子宮%母羊
아랍서림%FSH수체%단백표체%조직학결구%자궁%모양
alarelin%FSH receptor%protein expression%histostructure%uterus%ewe
为研究GnRHa主动免疫对绵羊子宫FSHR表达与分布及子宫发育的作用,探讨GnRHa调节生殖功能的机制,将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动物分别皮下注射阿拉瑞林( alarelin)抗原200、300和400μg,0 d和14 d各1次；EG-Ⅳ和EG-Ⅴ动物皮下注射alarelin抗原200μg和300μg,0、7、14和21 d各1次；对照组皮下注射药物溶媒,0 d和14 d各1次。于70 d在颈动脉放血处死绵羊,无菌切取两侧子宫角。免疫组织化学SP法染色并进行图像分析,光学显微镜和电子显微镜分别观察子宫显微和超微结构变化。 Western blot分析FSHR蛋白表达。结果发现各试验组FSHR蛋白表达量随着alarelin免疫剂量和次数的增多而逐渐增加。 EG-Ⅲ和EG-Ⅴ极显著高于对照组(P<0.01)。 FSHR主要分布于子宫内膜细胞和子宫腺上皮细胞的胞质和胞核；EG-Ⅰ、EG-Ⅱ和EG-Ⅲ灰度值逐渐升高,EG-Ⅲ和EG-V(P<0.01)显著高于CG,证明alarelin主动免疫能增强子宫FSHR的分布与表达,加大注射剂量和次数作用更明显。 EG-I、EG-Ⅱ和EG-Ⅲ的子宫质量、UWT和EET均变小,EG-Ⅲ和EG-Ⅴ显著小于对照组(P<0.05)。试验组出现不同程度的子宫壁和上皮变薄,细胞胞质减少,腺体数量减少,腺腔缩小,平滑肌细胞及核均变小。线粒体和线粒体嵴减少,微绒毛变短,以EG-Ⅲ最明显。结果表明,Alarelin主动免疫母羊能增强子宫FSHR的分布与蛋白表达,使子宫超微和超微结构明显改变,抑制子宫发育,尤其是子宫内膜的生长显著受阻。
為研究GnRHa主動免疫對綿羊子宮FSHR錶達與分佈及子宮髮育的作用,探討GnRHa調節生殖功能的機製,將42隻5~6月齡母綿羊(Ovis aries)隨機分為6組(n =7)。 EG-Ⅰ、EG-Ⅱ和EG-Ⅲ動物分彆皮下註射阿拉瑞林( alarelin)抗原200、300和400μg,0 d和14 d各1次；EG-Ⅳ和EG-Ⅴ動物皮下註射alarelin抗原200μg和300μg,0、7、14和21 d各1次；對照組皮下註射藥物溶媒,0 d和14 d各1次。于70 d在頸動脈放血處死綿羊,無菌切取兩側子宮角。免疫組織化學SP法染色併進行圖像分析,光學顯微鏡和電子顯微鏡分彆觀察子宮顯微和超微結構變化。 Western blot分析FSHR蛋白錶達。結果髮現各試驗組FSHR蛋白錶達量隨著alarelin免疫劑量和次數的增多而逐漸增加。 EG-Ⅲ和EG-Ⅴ極顯著高于對照組(P<0.01)。 FSHR主要分佈于子宮內膜細胞和子宮腺上皮細胞的胞質和胞覈；EG-Ⅰ、EG-Ⅱ和EG-Ⅲ灰度值逐漸升高,EG-Ⅲ和EG-V(P<0.01)顯著高于CG,證明alarelin主動免疫能增彊子宮FSHR的分佈與錶達,加大註射劑量和次數作用更明顯。 EG-I、EG-Ⅱ和EG-Ⅲ的子宮質量、UWT和EET均變小,EG-Ⅲ和EG-Ⅴ顯著小于對照組(P<0.05)。試驗組齣現不同程度的子宮壁和上皮變薄,細胞胞質減少,腺體數量減少,腺腔縮小,平滑肌細胞及覈均變小。線粒體和線粒體嵴減少,微絨毛變短,以EG-Ⅲ最明顯。結果錶明,Alarelin主動免疫母羊能增彊子宮FSHR的分佈與蛋白錶達,使子宮超微和超微結構明顯改變,抑製子宮髮育,尤其是子宮內膜的生長顯著受阻。
위연구GnRHa주동면역대면양자궁FSHR표체여분포급자궁발육적작용,탐토GnRHa조절생식공능적궤제,장42지5~6월령모면양(Ovis aries)수궤분위6조(n =7)。 EG-Ⅰ、EG-Ⅱ화EG-Ⅲ동물분별피하주사아랍서림( alarelin)항원200、300화400μg,0 d화14 d각1차；EG-Ⅳ화EG-Ⅴ동물피하주사alarelin항원200μg화300μg,0、7、14화21 d각1차；대조조피하주사약물용매,0 d화14 d각1차。우70 d재경동맥방혈처사면양,무균절취량측자궁각。면역조직화학SP법염색병진행도상분석,광학현미경화전자현미경분별관찰자궁현미화초미결구변화。 Western blot분석FSHR단백표체。결과발현각시험조FSHR단백표체량수착alarelin면역제량화차수적증다이축점증가。 EG-Ⅲ화EG-Ⅴ겁현저고우대조조(P<0.01)。 FSHR주요분포우자궁내막세포화자궁선상피세포적포질화포핵；EG-Ⅰ、EG-Ⅱ화EG-Ⅲ회도치축점승고,EG-Ⅲ화EG-V(P<0.01)현저고우CG,증명alarelin주동면역능증강자궁FSHR적분포여표체,가대주사제량화차수작용경명현。 EG-I、EG-Ⅱ화EG-Ⅲ적자궁질량、UWT화EET균변소,EG-Ⅲ화EG-Ⅴ현저소우대조조(P<0.05)。시험조출현불동정도적자궁벽화상피변박,세포포질감소,선체수량감소,선강축소,평활기세포급핵균변소。선립체화선립체척감소,미융모변단,이EG-Ⅲ최명현。결과표명,Alarelin주동면역모양능증강자궁FSHR적분포여단백표체,사자궁초미화초미결구명현개변,억제자궁발육,우기시자궁내막적생장현저수조。
The experiment was undertaken to evaluate the effects of active immunization against the gonado-tropin releasing hormone agonist( GnRHa) on the expression and immuno-localization of FSHR in uteri,as well as uterine development in the ewes. Forty two ewes ( Ovis aries) ,five-to-six months old,were randomly assigned into six groups (n=7). The animals in the experimental groups EG-Ⅰ,EG-Ⅱand EG-Ⅲwere subcutaneously injec-ted with 200 μg,300 μg and 400 μg alarelin antigens twice (on day 0 and 14),respectively,in groups EG-Ⅳand EG-Ⅴ were subcutaneously injected with 200 μg and 300 μg alarelin antigens four times ( on day 0 ,7 ,14 and 21),respectively and in control group (CG) were injected subcutaneously with 2. 0 mL solvent twice (on day 0 and 7 ) . The samples of the uterine horns of each ewe were collected aseptically on day 70 after killed by bloodletting from carotid. The FSHR distribution in the uteri was studied by the methods of Immunohistochemistry SP ( streptomyces avidin- peroxidase ) and image analysis, the mirostructure and ultrastructure changes of the uteri were observed under the optical and electron microscope,respectively and the expression of FSHR was de-tected by Western blot. The results showed that the expression of FSHR increased with the increasing of alarelin dose and injection times and the expression levels of FSHR in EG-Ⅲ and EG-Ⅴ were significantly higher than ( P<0. 01) in CG. The FSHR were predominantly concentrated in the cytoplasm and nuclei of the uterine endom-etrial cells and glandular epithelium cells. The gray values of microscopy images in EG-Ⅰ,EG-Ⅱand EG-Ⅲin-creased gradually and were significantly higher in EG-Ⅲand EG-Ⅴthan in CG (P<0. 01). The uterine weights, endometrial epithelium thickness ( EET) and uterine wall thickness ( UWT) in all experimental groups reduced and were significantly lower in EG-Ⅲ and EG-Ⅴ than in CG ( P<0 . 05 ) . The uterine cavities and glandular lu-men volumes narrowed,and the uterine wall became clearly thinner in all experimental groups. The quantities of mitochondria and mitochondria cristaes decreased and the microvilli became shorter,especially in EG-Ⅲ. In con-clusion,alarelin active immunity enhanced distribution of FSHR,promoted the expression of FSHR protein in the uteri of the ewes,changed the mirostructure and ultrastructure of the uteri,inhibited the uterine development of e-wes,and especially suppressed the growth of uterine endometrium.
