本文主要研究内容
作者王俊丽（2019）在《小檗碱通过miRNA-18b调控猪体外受精胚胎发育的分子机制》一文中研究指出：体外受精胚胎的体外培养是胚胎体外生产技术的重要环节。胚胎质量直接影响胚胎发育和胎儿生长。本研究首先观察了小檗碱(Berberine,Ber)和miRNA-18b对猪体外受精胚胎发育率、囊胚细胞增殖及凋亡的影响;其次,探讨Ber和miRNA-18b对猪体外受精胚胎内miRNA-18b、IGF-I、IGF-IR和HIF-1α在转录和翻译水平的调控作用,以及检测miRNA-18b的直接靶基因是否为IGF-I、IGF-IR和HIF-1α;最后探讨了Ber和miRNA-18b对AKT通路的调控关系,为提高猪体外受精胚胎体外发育提供理论及数据支持。试验以未添加Ber的NCSU-23胚胎基础培养液为对照组,Ber加入NCSU-23培养液中为Ber组,在对照组的原核胚细胞质中显微注射miRNA-18b过表达剂(agomir)、低表达剂(antagomir)和阴性片段分别为miRNA-18b agomir组、miRNA-18b antagomir组和NC组。观察IVF胚胎2-、4-、8-细胞和桑椹胚、囊胚发育率;囊胚细胞差异染色法、Hoechst33342-TUNEL法分别观察内细胞团和滋养层细胞数、细胞总数和囊胚细胞凋亡数;通过qRT-PCR、Western-blot和双荧光素酶报告基因的方法验证miRNA-18b的靶基因IGF-I、IGF-IR和HIF-1α;Western-blot检测AKT磷酸化水平。结果:Ber和miRNA-18b antagomir,可显著提高猪IVF胚胎发育率、囊胚内细胞团、滋养层、总细胞数和和细胞总数以及内细胞团细胞占细胞总数比率(P<0.01或P<0.05),极显著降低囊胚细胞凋亡数(P<0.01),提高了猪IVF胚胎的发育质量。在4-、8-细胞和桑椹胚期,Ber极显著地下调miRNA-18b的水平,而且miRNA-18b的水平随着胚胎发育呈下降趋势,4-~8-细胞时期更为明显;在4-、8-细胞和桑椹胚期,Ber和miRNA-18b antagomir能显著上调猪IVF胚胎中IGF-I、IGF-IR和HIF-1α表达水平(P<0.01或P<0.05),miRNA-18b agomir显著下调IGF-I、IGF-IR和HIF-1α表达水平(P<0.01或P<0.05);IGF-I、IGF-IR和HIF-1α的表达水平随着胚胎发育呈上升趋势,4-~8-细胞阶段更为明显;双荧光素酶报告基因结果表明在IGF-I、IGF-IR和HIF-1α的3’UTR wt、mut重建载体转染产生的双萤光素酶活性上,miRNA-18b与NC组均无显著差异(P>0.05)。Ber和miRNA-18b antagomir可极显著提高AKT磷酸化水平(P<0.01),miRNA-18b agomir能下调AKT磷酸化水平;AKT磷酸化水平随着胚胎发育呈上升趋势,4-~8-细胞阶段更为明显。结论:Ber能够通过下调4-、8-细胞和桑椹胚时期miRNA-18b的转录水平,促进IGF-I、IGF-IR和HIF-1α的表达,进而促进AKT的磷酸化,促进猪IVF胚胎细胞的增殖和体外发育,其中4-~8-细胞时期作用更为明显。然而,有关猪IVF胚胎中miRNA-18b的直接靶基因的问题还有待今后进一步的研究验证。
Abstract
ti wai shou jing pei tai de ti wai pei yang shi pei tai ti wai sheng chan ji shu de chong yao huan jie 。pei tai zhi liang zhi jie ying xiang pei tai fa yo he tai er sheng chang 。ben yan jiu shou xian guan cha le xiao bo jian (Berberine,Ber)he miRNA-18bdui zhu ti wai shou jing pei tai fa yo lv 、nang pei xi bao zeng shi ji diao wang de ying xiang ;ji ci ,tan tao Berhe miRNA-18bdui zhu ti wai shou jing pei tai nei miRNA-18b、IGF-I、IGF-IRhe HIF-1αzai zhuai lu he fan yi shui ping de diao kong zuo yong ,yi ji jian ce miRNA-18bde zhi jie ba ji yin shi fou wei IGF-I、IGF-IRhe HIF-1α;zui hou tan tao le Berhe miRNA-18bdui AKTtong lu de diao kong guan ji ,wei di gao zhu ti wai shou jing pei tai ti wai fa yo di gong li lun ji shu ju zhi chi 。shi yan yi wei tian jia Berde NCSU-23pei tai ji chu pei yang ye wei dui zhao zu ,Berjia ru NCSU-23pei yang ye zhong wei Berzu ,zai dui zhao zu de yuan he pei xi bao zhi zhong xian wei zhu she miRNA-18bguo biao da ji (agomir)、di biao da ji (antagomir)he yin xing pian duan fen bie wei miRNA-18b agomirzu 、miRNA-18b antagomirzu he NCzu 。guan cha IVFpei tai 2-、4-、8-xi bao he sang shen pei 、nang pei fa yo lv ;nang pei xi bao cha yi ran se fa 、Hoechst33342-TUNELfa fen bie guan cha nei xi bao tuan he zi yang ceng xi bao shu 、xi bao zong shu he nang pei xi bao diao wang shu ;tong guo qRT-PCR、Western-blothe shuang ying guang su mei bao gao ji yin de fang fa yan zheng miRNA-18bde ba ji yin IGF-I、IGF-IRhe HIF-1α;Western-blotjian ce AKTlin suan hua shui ping 。jie guo :Berhe miRNA-18b antagomir,ke xian zhe di gao zhu IVFpei tai fa yo lv 、nang pei nei xi bao tuan 、zi yang ceng 、zong xi bao shu he he xi bao zong shu yi ji nei xi bao tuan xi bao zhan xi bao zong shu bi lv (P<0.01huo P<0.05),ji xian zhe jiang di nang pei xi bao diao wang shu (P<0.01),di gao le zhu IVFpei tai de fa yo zhi liang 。zai 4-、8-xi bao he sang shen pei ji ,Berji xian zhe de xia diao miRNA-18bde shui ping ,er ju miRNA-18bde shui ping sui zhao pei tai fa yo cheng xia jiang qu shi ,4-~8-xi bao shi ji geng wei ming xian ;zai 4-、8-xi bao he sang shen pei ji ,Berhe miRNA-18b antagomirneng xian zhe shang diao zhu IVFpei tai zhong IGF-I、IGF-IRhe HIF-1αbiao da shui ping (P<0.01huo P<0.05),miRNA-18b agomirxian zhe xia diao IGF-I、IGF-IRhe HIF-1αbiao da shui ping (P<0.01huo P<0.05);IGF-I、IGF-IRhe HIF-1αde biao da shui ping sui zhao pei tai fa yo cheng shang sheng qu shi ,4-~8-xi bao jie duan geng wei ming xian ;shuang ying guang su mei bao gao ji yin jie guo biao ming zai IGF-I、IGF-IRhe HIF-1αde 3’UTR wt、mutchong jian zai ti zhuai ran chan sheng de shuang ying guang su mei huo xing shang ,miRNA-18byu NCzu jun mo xian zhe cha yi (P>0.05)。Berhe miRNA-18b antagomirke ji xian zhe di gao AKTlin suan hua shui ping (P<0.01),miRNA-18b agomirneng xia diao AKTlin suan hua shui ping ;AKTlin suan hua shui ping sui zhao pei tai fa yo cheng shang sheng qu shi ,4-~8-xi bao jie duan geng wei ming xian 。jie lun :Berneng gou tong guo xia diao 4-、8-xi bao he sang shen pei shi ji miRNA-18bde zhuai lu shui ping ,cu jin IGF-I、IGF-IRhe HIF-1αde biao da ,jin er cu jin AKTde lin suan hua ,cu jin zhu IVFpei tai xi bao de zeng shi he ti wai fa yo ,ji zhong 4-~8-xi bao shi ji zuo yong geng wei ming xian 。ran er ,you guan zhu IVFpei tai zhong miRNA-18bde zhi jie ba ji yin de wen ti hai you dai jin hou jin yi bu de yan jiu yan zheng 。
论文参考文献
论文详细介绍
论文作者分别是来自北京农学院的王俊丽，发表于刊物北京农学院2019-07-10论文，是一篇关于小檗碱论文,体外受精胚胎论文,体外发育论文,北京农学院2019-07-10论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自北京农学院2019-07-10论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：小檗碱论文; 体外受精胚胎论文; 体外发育论文; 北京农学院2019-07-10论文;