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. 2020 May 12;8:269. doi: 10.3389/fcell.2020.00269

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Copyright © 2020 Lin, Qu, Wang, Yang, Hu, Hu, Wu, Jiang, Liu and Han.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Morphine-induced ER stress enhances PKA-mediated phosphorylation of NMDA receptor. Representative western blot images were shown in this figure. (A,B) Morphine induced the phosphorylation of PKA and NR-1 in SH-SY5Y cells. Cells were collected 12 h after morphine (200 μM) treatment and analyzed by western blot (n = 3). (A,B) data were analyzed by Student’s t test (**P < 0.01, ***P < 0.001 vs. control). (C–E) Pretreatment with TUDCA could inhibit the up-regulation of ER stress-related molecule IRE1α, XBP1s and Caspase-12 induced by morphine in SH-SY5Y cells. TUDCA also suppressed the phosphorylation of PKA and NR-1 induced by morphine in SH-SY5Y cells. TUDCA (500 μM) was given 15 min before morphine (200 μM) administration. Cells were collected 12h after morphine treatment and analyzed by western blot (n = 3). (F–H) AEBSF inhibited the up-regulation of XBP1s induced by morphine in SH-SY5Y cells. AEBSF also suppressed the phosphorylation of PKA and NR-1induced by morphine in SH-SY5Y cells. AEBSF (200 μM) was given 15 min before morphine (200 μM) administration. Cells were collected 12 h after morphine treatment and analyzed by western blot (n = 3). (I–K) 4μ8C decreased the level of XBP1s induced by morphine in SH-SY5Y cells. 4μ8C also inhibited the phosphorylation of PKA and NR-1 induced by morphine in SH-SY5Y cells. 4μ8C (IRE1α inhibitor, 10 μM) was given 15 min before morphine (200 μM, 12 h) administration. Cells were collected 12 h after morphine treatment and analyzed by western blot (n = 3). (C–K) data were analyzed by one-way ANOVA (*P < 0.05, **P < 0.01, ***P < 0.001 vs. control, ^#P < 0.05, ^##P < 0.01 vs. morphine-treated group).