Objective. The present study selected PC12 cells to construct a neuronal injury model induced by glucocorticoids (GC) in vitro, aiming to explore whether the endoplasmic reticulum stress (ERS) PKR-like endoplasmic reticulum kinase (PERK)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) and inositol requirement 1 (IRE1)-apoptosis signal regulating kinase 1 (ASK1)-C-Jun amino-terminal kinase (JNK) signaling pathways are associated with the neuronal injury process induced by GC and provide morphological evidence. Methods. Cell models with different doses and different durations of GC exposure were established. The viability of PC12 cells was detected by the CCK-8 assay, and the apoptosis rate of PC12 cells was detected by the flow cytometry assay. The expression of microtubule-associated protein 2 (Map2); glucocorticoids receptor (GR); cellular oncogene fos (C-fos); and ERS-related proteins, glucose-regulated protein 78 (GRP78), p-PERK, p-IRE1, ATF4, ASK1, JNK, and CHOP, was observed by immunofluorescence staining. Results. The results of immunofluorescence staining showed that PC12 cells abundantly expressed Map2 and GR. The CCK-8 assay revealed that high-concentration GC exposure significantly inhibited the cell viability of PC12 cells. The flow cytometry assay indicated that high-concentration GC exposure significantly increased the apoptosis rate of PC12 cells. Immunofluorescence staining showed that GC exposure significantly increased the expression of C-fos, GRP78, p-PERK, p-IRE1, ATF4, ASK1, JNK, and CHOP. Treatment with ERS inhibitor 4-phenylbutyric acid (4-PBA) and GR inhibitor RU38486 attenuated related damage and downregulated the expression of the abovementioned proteins. Conclusion. High-concentration GC exposure can significantly inhibit the viability of PC12 cells and induce apoptosis. PERK-ATF4-CHOP and IRE1-ASK1-JNK pathways are involved in the above damage process.

中文翻译：

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内质网应激与糖皮质激素诱导的 PC12 细胞凋亡有关

客观。本研究选取PC12细胞构建体外糖皮质激素（GC）诱导的神经元损伤模型，旨在探讨内质网应激（ERS）PKR样内质网激酶（PERK）激活转录因子4（ATF4） C/EBP-同源蛋白 (CHOP) 和肌醇需求 1 (IRE1)-凋亡信号调节激酶 1 (ASK1)-C-Jun 氨基末端激酶 (JNK) 信号通路与 GC 诱导的神经元损伤过程相关，并提供形态学证据。方法. 建立了不同剂量和不同GC暴露时间的细胞模型。CCK-8法检测PC12细胞活力，流式细胞仪检测PC12细胞凋亡率。微管相关蛋白2（Map2）的表达；糖皮质激素受体（GR）；细胞癌基因 fos (C-fos)；免疫荧光染色观察 ERS ​​相关蛋白、葡萄糖调节蛋白 78 (GRP78)、p-PERK、p-IRE1、ATF4、ASK1、JNK 和 CHOP。结果. 免疫荧光染色结果显示PC12细胞大量表达Map2和GR。CCK-8 测定显示，高浓度 GC 暴露显着抑制 PC12 细胞的细胞活力。流式细胞仪检测表明，高浓度 GC 暴露显着增加了 PC12 细胞的凋亡率。免疫荧光染色显示，GC 暴露显着增加了 C-fos、GRP78、p-PERK、p-IRE1、ATF4、ASK1、JNK 和 CHOP 的表达。用 ERS ​​抑制剂 4-苯基丁酸 (4-PBA) 和 GR 抑制剂 RU38486 治疗可减轻相关损伤并下调上述蛋白质的表达。结论. 高浓度 GC 暴露可显着抑制 PC12 细胞的活力并诱导细胞凋亡。PERK-ATF4-CHOP 和 IRE1-ASK1-JNK 通路参与上述损伤过程。