玉米赤霉烯酮对断奶小母猪子宫形态学及热应激蛋白70分布和表达的影响  

Effects of Zearalenone on Expression of Heat Shock Protein 70 and Morphology of Uterus Tissues of Post-Weaning Piglets

作　　者：周敏[1] 周雪梅[1] 杨立杰[1] 黄丽波[1] 冯蕾 邵明慧 杨晨 杨维仁[1] 杨在宾[1] 姜淑贞[1] ZHOU Min;ZHOU XueMei;YANG LiJie;HUANG LiBo;FENG Lei;SHAO MingHui;YANG Chen;YANG WeiRen;YANG ZaiBin;JIANG ShuZhen(Shandong Provincial Laboratory of Animal Biotechnology and Disease Control and Province, College of Animal Science andTechnology, Shandong Agricultural University, Tai' an 271018, Shandong)

机构地区：[1]山东农业大学动物科技学院/山东省动物工程与疾病防治重点实验室,山东泰安271018

出　　处：《中国农业科学》2018年第4期778-788,共11页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31572441);山东省自然科学基金(ZR2017MC049);山东省现代农业产业技术体系生猪创新团队建设项目(SDAIT-08-04);"双一流"奖补资金资助

摘　　要：【目的】研究不同水平玉米赤霉烯酮(zearalenone,ZEA)对断奶小母猪子宫形态学和子宫内热应激蛋白70(heat shock protein 70,HSP70)的阳性分布规律及m RNA相对表达量的影响。【方法】选择25—28日龄健康三元杂交(杜×长×大)断奶小母猪40头,产床上适应环境10 d后转入单体笼(0.48 m2)。根据日龄(35—38)和平均体重(14.01±0.86 kg),将40头试验小母猪分为4个处理,每个处理10个重复,每个重复1头猪。对照组(Control)饲喂基础饲粮,处理1(ZEA0.5)、处理2(ZEA1.0)和处理3(ZEA1.5)分别在基础饲粮基础上添加0.5、1.0和1.5 mg·kg^(-1)的ZEA(ZEA的测定值分别为0、0.52±0.07、1.04±0.03和1.51±0.13 mg·kg^(-1)),小母猪单笼饲养,自由采食和饮水,预饲期10 d,正式期35 d。试验结束前一天对小母猪禁食12 h,全部放血屠宰,打开腹腔后无菌条件下迅速切取一份子宫组织置于2 m L无RNA酶的冻存管中液氮速冻,而后转至-80℃冻存,检测HSP70的m RNA相对表达量,另切取子宫组织块迅速固定于Bouin’s液,待做组织切片检测子宫形态学的结构和HSP70的阳性分布规律。【结果】随着饲粮中ZEA水平的升高,子宫肌层和内膜明显增厚,子宫腺数量明显增多,子宫腺密度明显增大;子宫肌层和内膜厚度随着饲粮中ZEA水平的升高呈一次(P<0.001)和二次(P<0.001)线性升高,1.5 mg·kg^(-1)处理的子宫肌层和内膜厚度显著高于1.0 mg·kg^(-1)处理(P<0.05),1.0 mg·kg^(-1)处理显著高于0.5 mg·kg^(-1)处理(P<0.05),0.5 mg·kg^(-1)处理又显著高于对照组(P<0.05)。免疫组化结果显示,HSP70免疫阳性物质主要分布于子宫肌层平滑肌细胞、子宫腺上皮细胞、腔上皮细胞和血管内皮细胞的胞质内,偶见胞核着色。对照组免疫阳性较弱呈浅黄色,随着饲粮中ZEA水平的升高,HSP70免疫阳性反应明显增强,颜色加深,呈黄色和棕黄色块状分布。断奶小母猪子宫中肌层、腺上皮、腔上皮及所测部分总HSP70免�【Objective】The aim of this study was to investigate the effects of zearalenone(ZEA)on uterus morphology,distribution and m RNA relative expression of heat shock protein 70(HSP 70)in post-weaning piglets.【Method】A total of 40 healthy female piglets(Duroc×Landrace×Large White)aged approximately 25-28 d were selected for study.The piglets were transferred to single cages after 10 day adaptation on the obstertric table,then they were allocated into four treatments.Each treatment consisted of 10 replicates of 1 pig per replicate according to the age(35-38)and the average body weight(14.01±0.86 kg).After 10 d of adaptation piglets were fed a normal basal diet supplemented with purified ZEA of 0(Control),0.5(0.52±0.07),1.0(1.04±0.03)and 1.5(1.51±0.13)mg·kg-1 for 35 d.Animals were housed individually with free access to feed and water.At the end of the feeding trial,piglets were deprived with feed for 12 h and sacrificed.Uterus were then isolated under sterile conditions.Two samples of uterus tissues from each pig were quickly collected,one of which was stored at-80℃to detect relative m RNA expression of HSP70,and the other was fixed promptly in Bouin’s for histological examination and immunohistochemical analysis.【Result】Results showed that the myometrium and endometrium became thicker and the number and density of uterine glands increased with the higher level of ZEA.The thickness of myometrium and endometrium increased linearly(P<0.001)and quadratically(P<0.001)with higher level of ZEA.The thickness of myometrium and endometrium from ZEA1.5,1.0 and 0.5 treatments were significantly higher than that of ZEA1.0,0.5 and control treatments,respectively(P<0.05).The results of immunohistochemistry indicated that HSP70 immunoreactive substance was mainly distributed in the cytoplasm of uterine smooth muscle cells,uterine glandular epithelial cells,luminal epithelial cells and vascular endothelial cells,but nuclear staining was rarely observed.The light yellow immunoreactive substance was shown in the ti

关 键 词：玉米赤霉烯酮 断奶小母猪 子宫 热应激蛋白70 

分 类 号：S859.8[农业科学—临床兽医学]