摘要
为初步探究内源性绵羊肺腺瘤病毒(en JSRV)囊膜蛋白(Env)在绵羊绒毛膜滋养层细胞中的作用机制,利用免疫组织化学染色法检测enJSRV-Env在绵羊胎盘中的主要表达区域,继而瞬时转染重组质粒pEGFP-C1/en JSRV-env到绵羊绒毛膜滋养层细胞,并加入Erk抑制因子PD 98059、p38抑制因子SB 203580及JN K抑制因子SP600125,同时转染重组质粒至293T细胞中作为对比。利用W estern-blot分别检测2种细胞中Erk1/2、p38和JNK蛋白激酶磷酸化水平的变化。免疫组织化学染色结果显示,enJSRV-Env主要表达于绵羊胎盘中绒毛膜滋养层细胞。Western-blot结果显示,绵羊绒毛膜滋养层细胞中,转染重组质粒pEGFP-C1/en JSRV-env组中p-Erk1/2、p-p38和p-JNK的表达量均显著高于对照组和加入抑制因子组(P<0.05);加入3种抑制因子后,3种蛋白激酶的磷酸化水平较转染组显著降低。293T细胞中,转染重组质粒pEGFP-C1/en JSRVenv组中p-p38的表达量显著高于对照组(P<0.05),p-JNK的表达量极显著高于对照组(P<0.01)。因此,enJSRV-Env激活了绵羊绒毛膜滋养层细胞中MAPK信号转导通路。再通过与293T细胞的结果对比可知,enJSRV-Env在这2种细胞中的作用机制有所不同。本研究为今后继续深入探究en JSRV-Env的作用机制提供了新的思路和依据。
To explore role of endogenous jaagsiekte sheep retrovirus（en JSRV） envelope protein（Env）in sheep chorionic trophoblast cells,we detected the main positive regions of en-JSRV-Env expression in the placenta of sheep by immunohistochemistry.Then the recombinant plasmid p EGFP-C1/en JSRV-env was transiently transfected into sheep chorionic trophoblast cells,treated with Erk inhibitor PD98059,p38 inhibitor SB203580 and JNK inhibitor SP600125.The recombinant plasmid was transiently transfected into 293 T cells for comparison.Then we detected the levels of p Erk1/2,p-p38 and p-JNK in sheep chorionic trophoblast cells and 293 T cells by Western-blot,respectively.Immunohistochemistry results revealed that en JSRV-Env was mainly expressed in chorionic trophoblast cells of the placenta.West-ern-blot results showed that the levels of p-Erk1/2,p-p38 and p-JNK in sheep chorionic trophoblast cells with recombinant plasmid were significantly higher than that in the control groups and the inhibitor groups（P〈0.05）.The level of p-p38 in 293 T cells with recombinant plasmid was significantly higher than that in the control group（P〈0.05）,and the levels of p-JNK was extremely significantly higher than that in the control group（P〈0.01）.We concluded that en JSRV-Env activated MAPK signaling pathway in sheep chorionic trophoblast cells.The role of en JSRV-Env was different in the two cell lines.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第8期1043-1050,共8页
Chinese Veterinary Science
基金
国家自然科学基金项目(31360597)
内蒙古科技厅应用研究项目(201502070)
内蒙古草原英才创新团队项目(20151031)
