小鼠脑组织及培养神经元Neuro-2a在内质网应激反应时的基因表达谱分析

发布时间：2018-06-16 22:29

本文选题：内质网应激反应 + 二代RNA测序　；参考：《中国生物化学与分子生物学报》2017年04期

【摘要】：内质网是真核细胞的重要细胞器。某些细胞内外因素如病原体感染等能引起从内质网到胞浆和胞核的信号传导途径活化,即内质网应激反应。但是,目前国内外尚无针对内质网应激反应的基因表达谱分析报道。本研究中,用3种已报道的内质网应激反应诱导剂,包括蛋白质糖基化抑制剂衣霉素(tunicamycin)、内质网Ca~(2+)-ATPases抑制剂毒胡萝卜素(thapsigargin)和乙脑病毒(Japanese encephalitis virus,JEV),分别处理小鼠颅腔和小鼠脑神经瘤细胞(Neuro-2a),试剂处理组与未处理组的第二代RNA测序分析发现,衣霉素、毒胡萝卜素和乙脑病毒在体外和体内均引起分子伴侣基因Hsp70表达上调,诱导内质网应激反应。衣霉素、毒胡萝卜素和乙脑病毒体外处理诱导的内质网应激反应信号通路中,基因差异表达相似性高于体内处理组。乙脑病毒和糖基化抑制剂衣霉素体内外处理,主要诱导内质网应激反应的非折叠蛋白质反应信号通路,引起相关基因Atf4、Bip、Edem和Perk等表达上调。内质网Ca~(2+)-ATPases抑制剂毒胡萝卜素主要诱导内质网超负荷反应,激活NF-κB信号通路。乙脑病毒诱导的内质网应激反应相关差异表达基因数量最多,体外与体内合计有40种。乙脑病毒体内外处理上调的基因包括Bax、Casp12、Atf4、Bip、Edem和Perk等,下调的基因包括Sec23/24、Nef、Svip和Jnk等。糖基化抑制剂衣霉素体内外处理上调基因包括Gadd34、Atf4、Ermani和Bip等,下调基因包括Grp94、Atf6、Sec23/24和Nef等。内质网Ca2+-ATPases抑制剂毒胡萝卜素体内外处理上调的基因包括Sec61、Trap和Ask1等。衣霉素、毒胡萝卜素和乙脑病毒体内外处理也通过内质网应激反应,调控与炎症或凋亡相关的MAPK信号通路和P53信号通路。本研究首次通过使用3种内质网应激反应诱导剂分别处理小鼠和细胞,揭示了体内外内质网应激反应引起的基因表达谱变化,为内质网应激反应相关疾病的治疗提供了新思路。
[Abstract]:Endoplasmic reticulum is an important organelle of eukaryotic cells. Some internal and external factors, such as pathogen infection, can activate the signal transduction pathway from endoplasmic reticulum to cytoplasm and nucleus, that is, endoplasmic reticulum stress reaction. However, there is no analysis of endoplasmic reticulum stress response gene expression profile at home and abroad. In this study, three reported endoplasmic reticulum stress inducers were used. The protein glycosylation inhibitor, tunicamycin, the endoplasmic reticulum (ER) and the Japanese encephalitis virus (JEV) were used to treat the cranial cavity of mice and the neuro-2aanine of brain neuroma cells, respectively. The second generation RNA sequencing of the reagents treated group and the untreated group was found. In vitro and in vivo, the expression of the molecular chaperone gene Hsp70 was up-regulated and the endoplasmic reticulum stress was induced by itamycin, carotene and Japanese encephalitis virus. The similarity of gene differential expression in the endoplasmic reticulum (ER) stress response signaling pathway induced by itamycin, carotene and encephalitis B virus in vitro was higher than that in vivo treatment group. In vivo and in vitro treatment with Japanese encephalitis B virus and glycosylation inhibitor itamycin mainly induces the signal pathway of non-folded protein reaction in endoplasmic reticulum stress reaction and up-regulates the expression of the related genes Atf4- Bip-Edem and Perk. Endoplasmic reticulum (ER) caprostin-ATPase inhibitor, carotene, mainly induces endoplasmic reticulum overload and activates NF- 魏 B signaling pathway. The number of differentially expressed genes related to endoplasmic reticulum stress induced by encephalitis B virus was the highest, with 40 genes in vitro and in vivo. The up-regulated genes of Japanese encephalitis virus in vivo and in vitro include Baxon Casp12Atf4, BipEdem and Perk, while down-regulated genes include Sec23 / 24 NefSvip and Jnk. The up-regulated genes including Gadd34, Atf4, Ermani and Bip, and down-regulated genes include Grp94, Atf6, Sec23 / 24 and Nef, respectively. Endoplasmic reticulum Ca 2-ATPase inhibitor, carotene, up-regulated genes including Sec61Trap and Ask1 in vitro and in vivo. In vivo and in vitro treatment of itamycin, carotene and encephalitis B virus also regulate MAPK signaling pathway and p53 signaling pathway related to inflammation or apoptosis through endoplasmic reticulum stress. In this study, three endoplasmic reticulum stress inducers were used to treat mice and cells for the first time, which revealed the changes of gene expression profile induced by endoplasmic reticulum stress in vivo and in vitro, and provided a new idea for the treatment of endoplasmic reticulum stress related diseases.
【作者单位】：江西农业大学生物科学与工程学院;江西省疾病预防控制中心;南昌大学生命科学院人类衰老研究所;
【基金】：国家自然科学基金项目(No.31460667) 江西省重点项目(No.20161BBF60084)资助~~
【分类号】：R363

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