本文主要研究内容
作者邹晓琳（2019）在《EV71诱导人神经细胞SH-SY5Y自噬及凋亡的机制研究》一文中研究指出：目的:证明EV71具有诱导神经细胞自噬及凋亡的作用;初步探讨了EV71诱导神经细胞自噬的可能的分子机制。方法:在体外细胞实验中,运用芯片技术检测了人神经细胞SH-SY5Y感染EV71病毒后细胞内差异表达的miRNAs;在众多差异表达的miRNAs中,通过qRT-PCR验证了部分差异表达的基因,并初步确定了可能与自噬相关的mir-29b,运用生物信息学手段寻找到其可能的靶基因PLD1;同时运用qRT-PCR,western blot及激光共聚焦技术检测了感染EV71病毒的SH-SY5Y细胞发生自噬的情况;随后使用mir-29b mimics来调节细胞内mir-29b的表达水平后,再检测细胞自噬水平的变化情况;运用western blot及流式细胞术检测了病毒感染后细胞的凋亡情况。结果:qRT-PCR实验结果显示EV71可下调SH-SY5Y细胞mir-29b的表达水平,自噬相关的基因beclin-1表达水平明显上调,p62表达量则显著下降;western blot结果显示EV71感染细胞后,LC3-Ⅱ/LC3-Ⅰ比值增加;此外,随着感染时间推移,细胞内GFP-LC3荧光集聚显著增加。通过转染miRNA mimics上调细胞内mir-29b的表达后,检测EV71感染神经细胞后自噬相关基因表达的变化情况,其中Beclin-1表达量有所下调,p62表达量则明显上升,PLD1表达量出现明显上升;western blot中的自噬指标LC3-Ⅱ/LC3-Ⅰ的比值则有明显减小。western blot结果显示,随EV71感染细胞的时间推移Bcl-2的表达量逐渐下降;与此同时,流式细胞术检测结果表明随EV71感染时间的延长,发生凋亡的细胞数量显著增加。结论:上述实验结果提示,EV71可诱导SH-SY5Y细胞出现自噬及凋亡,下调细胞内的mir-29b表达;当上调细胞内mir-29b的表达水平时,EV71诱导SH-SY5Y自噬的作用可被逆转。说明EV71诱导人神经源性细胞SH-SY5Y发生自噬的作用是通过mir-29b途径实现的。
Abstract
mu de :zheng ming EV71ju you you dao shen jing xi bao zi shi ji diao wang de zuo yong ;chu bu tan tao le EV71you dao shen jing xi bao zi shi de ke neng de fen zi ji zhi 。fang fa :zai ti wai xi bao shi yan zhong ,yun yong xin pian ji shu jian ce le ren shen jing xi bao SH-SY5Ygan ran EV71bing du hou xi bao nei cha yi biao da de miRNAs;zai zhong duo cha yi biao da de miRNAszhong ,tong guo qRT-PCRyan zheng le bu fen cha yi biao da de ji yin ,bing chu bu que ding le ke neng yu zi shi xiang guan de mir-29b,yun yong sheng wu xin xi xue shou duan xun zhao dao ji ke neng de ba ji yin PLD1;tong shi yun yong qRT-PCR,western blotji ji guang gong ju jiao ji shu jian ce le gan ran EV71bing du de SH-SY5Yxi bao fa sheng zi shi de qing kuang ;sui hou shi yong mir-29b mimicslai diao jie xi bao nei mir-29bde biao da shui ping hou ,zai jian ce xi bao zi shi shui ping de bian hua qing kuang ;yun yong western blotji liu shi xi bao shu jian ce le bing du gan ran hou xi bao de diao wang qing kuang 。jie guo :qRT-PCRshi yan jie guo xian shi EV71ke xia diao SH-SY5Yxi bao mir-29bde biao da shui ping ,zi shi xiang guan de ji yin beclin-1biao da shui ping ming xian shang diao ,p62biao da liang ze xian zhe xia jiang ;western blotjie guo xian shi EV71gan ran xi bao hou ,LC3-Ⅱ/LC3-Ⅰbi zhi zeng jia ;ci wai ,sui zhao gan ran shi jian tui yi ,xi bao nei GFP-LC3ying guang ji ju xian zhe zeng jia 。tong guo zhuai ran miRNA mimicsshang diao xi bao nei mir-29bde biao da hou ,jian ce EV71gan ran shen jing xi bao hou zi shi xiang guan ji yin biao da de bian hua qing kuang ,ji zhong Beclin-1biao da liang you suo xia diao ,p62biao da liang ze ming xian shang sheng ,PLD1biao da liang chu xian ming xian shang sheng ;western blotzhong de zi shi zhi biao LC3-Ⅱ/LC3-Ⅰde bi zhi ze you ming xian jian xiao 。western blotjie guo xian shi ,sui EV71gan ran xi bao de shi jian tui yi Bcl-2de biao da liang zhu jian xia jiang ;yu ci tong shi ,liu shi xi bao shu jian ce jie guo biao ming sui EV71gan ran shi jian de yan chang ,fa sheng diao wang de xi bao shu liang xian zhe zeng jia 。jie lun :shang shu shi yan jie guo di shi ,EV71ke you dao SH-SY5Yxi bao chu xian zi shi ji diao wang ,xia diao xi bao nei de mir-29bbiao da ;dang shang diao xi bao nei mir-29bde biao da shui ping shi ,EV71you dao SH-SY5Yzi shi de zuo yong ke bei ni zhuai 。shui ming EV71you dao ren shen jing yuan xing xi bao SH-SY5Yfa sheng zi shi de zuo yong shi tong guo mir-29btu jing shi xian de 。
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论文详细介绍
论文作者分别是来自武汉科技大学的邹晓琳，发表于刊物武汉科技大学2019-07-16论文，是一篇关于自噬论文,凋亡论文,武汉科技大学2019-07-16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，资料来自武汉科技大学2019-07-16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：自噬论文; 凋亡论文; 武汉科技大学2019-07-16论文;