妊娠期和分娩前后牦牛胎盘组织形态学变化及细胞凋亡的研究  

Study on the Morphological Changes and Apoptosis of Yak(Bos grunniens) Placentomes from Pregnancy, Pre and Post-parturation

作　　者：韩小红[1,2] 樊江峰 于艺腾[2] 何翃闳 马悦 余四九 崔燕 HAN Xiao-Hong;FAN Jiang-Feng;YU Yi-Teng;HE Hong-Hong;MA Yue;YU Si-Jiu;CUI Yan(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep ＆ Goat,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]甘肃省牛羊胚胎工程技术研究中心,兰州730070

出　　处：《农业生物技术学报》2018年第10期1714-1722,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31660732);甘肃省杰出青年基金(No.1506RJDA002);甘肃省农业科技创新课题(No.GNCX-2014-30);甘肃农业大学动物医学院教研产学支持计划(No.JYCX-KXO12)

摘　　要：牦牛(Bos grunniens)具有典型的子叶型胎盘,在妊娠过程中经历了从形成、生长发育到成熟的变化,是妊娠期连接母体和胎儿的纽带,发挥着营养物质和代谢废物交换、激素分泌等重要的功能。为探讨细胞凋亡与胎盘不同时期主要生理功能之间的相互关系,本研究采集4~8岁分别处于正常妊娠期、分娩前后牦牛的胎盘组织,将其组织制成石蜡切片并进行苏木精-伊红染色(hematoxylin-eosin staining, HE)染色,观察妊娠各阶段胎盘结构的变化;通过末端脱氧核苷酸转移酶介导的dUTP切口及末端标记(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, TUNEL)技术和4,6-二脒基-2-苯基吲哚(4,6-diamidino-2-phenylindole, DAPI)染色技术观察和分析胎盘组织细胞凋亡的情况。结果显示:妊娠期牦牛胎盘的基本结构在形成过程中处于不断变化的状态,怀孕45 d前后时已形成典型的上皮绒毛膜型胎盘结构;随着胎儿的发育,胎儿绒毛分支并深入母体隐窝中,双核滋养细胞在该结构中的数量也逐渐增多。子宫隐窝上皮在妊娠6个月前后时开始出现退行性变化,如细胞空泡化,整个隐窝上皮到分娩前基本消失,细胞凋亡现象存在于整个妊娠期牦牛胎盘组织中。胎儿绒毛上皮、母体隐窝上皮是TUNEL阳性细胞主要的分布场所,而子宫肉阜基质在妊娠早期少有凋亡细胞存在。随着胎儿的发育,母体隐窝上皮和胎儿绒毛上皮中的TUNEL阳性细胞也逐渐增多,且母体部分明显低于胎儿部分。胎儿绒毛上皮中的阳性细胞数在分娩前显著升高并在分娩后达到最大。本研究从组织学水平上对胎盘发育过程中的形态学变化和凋亡特征进行研究,研究结果为进一步解决牦牛生产过程中出现的如流产等问题提供一定的理论依据。Yak（Bos grunniens） has a typical cotyledon placenta which undergoes changes from formation,growth, and maturation during pregnancy, and it is the link between maternal and fetal during pregnancy. It is play an important role in the exchange of nutrients and metabolic waste, hormone secretion. Studying on the relationship between the phenomenon of apoptosis and the main physiological function in the placentome is very improtant for analyzing the mechanism of dynamic development of yak placentome in the angle of apoptosis. Placenta tissues during pregnancy, pre and post parturation of 4 - 8-year-old yaks were collected.They were paraffin-embedded and HE dyeinged to observe the changes of placental structure in all stages of pregnancy. The apoptosis of yak placental tissue was observed and analyzed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling（TUNEL） and 4, 6-diamidino-2-phenylindole（DAPI）dyeing technology. The structure of yaks placentomes during pregnancy was constantly changing, so the different stage of gestation could present different morphological characteristics. Yaks had formed a typical structure of the epithelial chorionic placenta in 45 days of pregnancy. With the progress of pregnancy, fetal villi gradually branched and deeply into the maternal caruncula crypts. The number of binucleate cell was increased in placentomes gradually. Caruncula crypt epithelium in 6 months of pregnancy began to appear obviously degenerative changes, such as cell vacuolization. The entire crypt epithelium was not found at preparturition. Yak placentomes existed in obvious apoptosis phenomenon. Fetal villi epithelium and maternal crypt epithelium was the main distribution of TUNEL positive cells during gestation and uterine meat Fu matrix had a very small amount of apoptotic cells in early stage of gestation. With the grown of the fetus,TUNEL-positive cells in the two layer epithelial tissue generally showed an upward trend, and the maternal crypt epithelium was significantly

关 键 词：牦牛 胎盘 细胞凋亡 组织结构 末端脱氧核苷酸转移酶介导的dUTP切口及末端标记(TUNEL) 

分 类 号：S857.2[农业科学—临床兽医学]