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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。
Endoplasmic reticulum is an important organelle of eukaryotic cells. Certain intracellular and extracellular factors such as pathogen infection can cause activation of the signal transduction pathway from the endoplasmic reticulum to the cytoplasm and nucleus, ie, the endoplasmic reticulum stress response. However, at present, there is no report on the gene expression profile for ER stress response at home and abroad. In this study, three kinds of endoplasmic reticulum stress inducing agents, including protein glycosylation inhibitor tunicamycin, endoplasmic reticulum Ca ~ (2 +) - ATPase inhibitor thapsigargin ( thapsigargin and Japanese encephalitis virus (JEV) were used to treat mouse cranial and mouse brain neuroma cells (Neuro-2a) respectively. Superoxide dismutase and JE virus cause up-regulation of Hsp70, a chaperone gene in vitro and in vivo, and induce ER stress. In tuberculin, thapsigargin and JE virus induced endoplasmic reticulum stress response signal transduction pathway, the similarity of gene expression difference was higher than that of in vivo treatment group. JE virus and glycosylated inhibitor tunicamycin in vitro and in vivo, mainly induced the non-folded protein response signal pathway of endoplasmic reticulum stress, resulting in the up-regulation of related genes such as Atf4, Bip, Edem and Perk. Endoplasmic reticulum Ca ~ (2 +) - ATPase inhibitor thapsigargin induces endoplasmic reticulum overload reaction and activates NF-κB signaling pathway. Encephalitis virus induced endoplasmic reticulum stress response related genes expressed the largest number, in vitro and in vivo total of 40 species. Genes up-regulated by JE virus in vitro and in vivo include Bax, Casp12, Atf4, Bip, Edem and Perk, down-regulated genes include Sec23 / 24, Nef, Svip and Jnk. The up-regulated genes of tunicamycin, a glycosylation inhibitor, in vitro and in vivo, include Gadd34, Atf4, Ermani and Bip, down-regulated genes include Grp94, Atf6, Sec23 / 24 and Nef. Endoplasmic reticulum Ca2 + -ATPases inhibitors thapsigargin in vitro and up-regulated genes include Sec61, Trap and Ask1 and so on. Tunicamycin, thapsigargin and JE virus also regulate MAPK and P53 signaling pathways that are associated with inflammation or apoptosis through endoplasmic reticulum stress. In this study, we first treated mice and cells by using three kinds of endoplasmic reticulum stress inducing agents respectively, and revealed the changes of gene expression profile caused by endoplasmic reticulum stress reaction in vivo, which is the treatment of endoplasmic reticulum stress-related diseases Provide a new idea.