抗凋亡蛋白cFLIP在山羊妊娠黄体中的表达  

Expression of cellular FLICE-like inhibitory protein (cFLIP) in corpus luteum during pregnancy in goats

作　　者：王延莉 娄安钢[1] 金花子[1] 金海国[2] WANG Yan-li;LOU An-gang;JIN Huai-zi;JIN Hai-guo(College of Agronomy,YanbianUniversity,Yanji,Jilin 133000,China;Animal Husbandry Division,Jilin Academy of AgriculturalSciences,Gongzhuling,Jilin 136100,CAzna)

机构地区：[1]延边大学农学院,吉林延吉133OOO [2]吉林省农业科学院畜牧分院,吉林公主岭136100

出　　处：《中国兽医学报》2019年第8期1622-1627,1634,共7页Chinese Journal of Veterinary Science

基　　金：教育部留学回国人员科研启动基金资助项目(教外司留(2012)1707)

摘　　要：为研究山羊妊娠黄体中抗凋亡蛋白,即细胞FLICE样抑制蛋白(cellular FLICE-Like inhibitory protein,cFLIP)的表达,分析探讨cFLIP在维持山羊正常妊娠黄体中的作用。试验收集山羊不同妊娠阶段(19,31,92,142 d)的黄体组织,分别采用RT-PCR、Western blot及免疫组织化学分析等方法,以确定cFLIP免疫定位,检测cFLIP mRNA及其蛋白在山羊不同妊娠阶段黄体中的表达水平;分别以3-磷酸甘油醛脱氢酶(GAPDH)mRNA和β-肌动蛋白(β-actin)作为内在对照。结果显示,cFLIP短链(cFLIP_S)和cFLIP长链(cFLIP_L)的mRNA及cFLIP_S蛋白在妊娠期各阶段的表达水平无显著性差异(P>0.05),而妊娠31d的cFLIP_L蛋白的表达水平却显著低于其他妊娠组(P<0.05);免疫染色结果显示,cFLIP蛋白的阳性反应在山羊妊娠不同阶段的黄体中均有高度表达;在妊娠19,92,142 d的黄体中均有观察到PCNA阳性,在妊娠31 d发现PCNA阳性较微弱;并且在所有妊娠组黄体中,几乎观察不到细胞凋亡(TUNEL阳性)。因此,推测cFLIP作为一种生存因子,参与维持山羊正常妊娠,其在妊娠黄体组织中的高水平表达可能在维持山羊正常妊娠中发挥抗凋亡作用。In order to study the expression of anti-apoptotic protein in goat?s pregnancy corpus luteum,namely cellular FLICE-like inhibitory protein(cFLIP),analyze cFLIP role in maintaining normal pregnancy corpus luteum goats.In this experiment,the luteal tissues of goats at different stages of pregnancy(19,31,92 and 142 d) were collected,respectively using RT-PCR,Western blot and immunohistochemical analysis to determine the immunohistochemistry of cFLIP,and to detect the expression level of cFLIP mRNA and protein in the corpus luteum of goats at different stages of pregnancy.Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) mRNA and β-actin were used as internal controls.The results showed that there was no significant difference in the expression levels of cFLIP_S and cFLIP_L mRNA and cFLIP_S protein during pregnancy(P>0.05),while the expression level of cFLIP_L protein on the 31 st day of pregnancy was significantly lower than that of other pregnancy groups(P<0.05).Immunostaining showed that the positive reaction of cFLIP protein was highly expressed in the corpus luteum of goats at different stages of pregnancy.PCNA positivity was observed in corpus luteum on the 19,92 and 142 d of pregnancy,and PCNA positivity was found to be weak on the 31 d of pregnancy.And almost no apoptosis(TUNEL positive) was observed in the corpus luteum of pregnancy group.Therefore,it is speculated that cFLIP,as a survival factor,is involved in maintaining normal pregnancy of goats,and its high-level expression in the luteal tissue of pregnancy may play an anti-apoptotic role in maintaining normal pregnancy of goats.

关 键 词：山羊 细胞FLICE样抑制蛋白(cFLIP) 细胞凋亡 黄体 妊娠期 

分 类 号：S814.1[农业科学—畜牧学] S852.21[农业科学—畜牧兽医]