内质网应激通过PERK-ATF4-CHOP和IRE1-ASK1-JNK信号通路参与应激诱导的大鼠下丘脑神经元损伤。
Endoplasmic Reticulum Stress Is Involved in Stress-Induced Hypothalamic Neuronal Injury in Rats via the PERK-ATF4-CHOP and IRE1-ASK1-JNK Pathways.
作者信息
Yi Shanyong, Chen Ke, Zhang Lihua, Shi Weibo, Zhang Yaxing, Niu Shiba, Jia Miaomiao, Cong Bin, Li Yingmin
机构信息
Hebei Key Laboratory of Forensic Medicine, Collaborative Innovation Center of Forensic Medical Molecular Identification, Department of Forensic Medicine, Hebei Medical University, Shijiazhuang, China.
出版信息
Front Cell Neurosci. 2019 May 3;13:190. doi: 10.3389/fncel.2019.00190. eCollection 2019.
OBJECTIVE
As a high-level nerve center that regulates visceral and endocrine activity, the hypothalamus plays an important role in regulating the body's stress response. Previous studies have shown that stress can cause damage to hypothalamic neurons. The present study aimed to further clarify the mechanism of endoplasmic reticulum stress (ERS) involvement in hypothalamic neuronal injury.
METHODS
A 7-day stressed rat model was established with daily restraining for 8 h and forced ice-water swimming for 5 min. The rats were randomly divided into control, stress, stress + GSK2606414 (PERK phosphorylation inhibitor), stress + KIRA6 (IRE1 phosphokinase activity inhibitor), GSK2606414, and KIRA6 groups. The pathological changes of hypothalamic neurons were observed by thionine staining. Expression of ERS proteins GRP78, ATF4, ASK1, JNK, and CHOP in the hypothalamic neurons were observed by immunohistochemical staining. The expression of JNK and CHOP mRNA in the hypothalamic neurons were observed by RNA hybridization (RNA Scope) and the expression of related proteins and mRNA was semiquantitatively analyzed by microscopy-based multicolor tissue cytometry (MMTC).
RESULTS
Thionine staining revealed that stress exposure resulted in edema, a lack of Nissl bodies, and pyknosis in hypothalamic neurons. Immunohistochemistry and RNA Scope showed that stress exposure significantly increased the expression of GRP78, ATF4, ASK1, CHOP, JNK, JNK mRNA, and CHOP mRNA. Treatment with PERK and IRE1 inhibitors attenuated pathological damage and downregulated the expression of ATF4, ASK1, JNK, CHOP, JNK mRNA, and CHOP mRNA.
CONCLUSION
Stress caused pathological changes in rat hypothalamic neurons. ERS PERK-ATF4-CHOP and IRE1-ASK1-JNK pathways were involved in the injury process.
目的
下丘脑作为调节内脏和内分泌活动的高级神经中枢，在调节机体应激反应中发挥重要作用。以往研究表明，应激可导致下丘脑神经元损伤。本研究旨在进一步阐明内质网应激（ERS）参与下丘脑神经元损伤的机制。
方法
建立为期7天的应激大鼠模型，每天束缚8小时并强迫冰水游泳5分钟。将大鼠随机分为对照组、应激组、应激+GSK2606414（PERK磷酸化抑制剂）组、应激+KIRA6（IRE1磷酸激酶活性抑制剂）组、GSK2606414组和KIRA6组。用硫堇染色观察下丘脑神经元的病理变化。用免疫组织化学染色观察下丘脑神经元中ERS蛋白GRP78、ATF4、ASK1、JNK和CHOP的表达。用RNA杂交（RNA Scope）观察下丘脑神经元中JNK和CHOP mRNA的表达，并用基于显微镜的多色组织细胞术（MMTC）对相关蛋白和mRNA的表达进行半定量分析。
结果
硫堇染色显示，应激暴露导致下丘脑神经元水肿、尼氏体缺失和核固缩。免疫组织化学和RNA Scope显示，应激暴露显著增加GRP78、ATF4、ASK1、CHOP、JNK、JNK mRNA和CHOP mRNA的表达。用PERK和IRE1抑制剂处理可减轻病理损伤，并下调ATF4、ASK1、JNK、CHOP、JNK mRNA和CHOP mRNA的表达。
结论
应激导致大鼠下丘脑神经元发生病理变化ERS的PERK-ATF4-CHOP和IRE1-ASK1-JNK通路参与了损伤过程。
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