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High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways

Bupivacaine has previously been reported to induce neurotoxicity, which is further enhanced by high glucose levels. In the present study, the underlying molecular mechanisms via which bupivacaine induces cytotoxicity under high glucose conditions were investigated in cultured human SH-SY5Y cells. In...

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Autores principales: Liu, Yongzhe, Sun, Li, Ma, Yaqun, Wei, Biyu, Gao, Minglong, Shang, Lixin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6691238/
https://www.ncbi.nlm.nih.gov/pubmed/31524237
http://dx.doi.org/10.3892/mmr.2019.10524
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author Liu, Yongzhe
Sun, Li
Ma, Yaqun
Wei, Biyu
Gao, Minglong
Shang, Lixin
author_facet Liu, Yongzhe
Sun, Li
Ma, Yaqun
Wei, Biyu
Gao, Minglong
Shang, Lixin
author_sort Liu, Yongzhe
collection PubMed
description Bupivacaine has previously been reported to induce neurotoxicity, which is further enhanced by high glucose levels. In the present study, the underlying molecular mechanisms via which bupivacaine induces cytotoxicity under high glucose conditions were investigated in cultured human SH-SY5Y cells. In order to identify the optimal concentrations of glucose and bupivacaine that induced cytotoxicity, SH-SY5Y cells were treated with 30–100 mM glucose and 0.5–1.0 mM bupivacaine. Based on the dose response experiments, 50 mM glucose and 0.5 mM bupivacaine was used in the present study. The effects that 3-MA (autophagy inhibitor) and rapamycin (RAPA; autophagy inducer) exerted on cell apoptosis, autophagy and the expression of protein kinase R-like endoplasmic reticulum kinase (PERK)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) and inositol-requiring enzyme 1 (IRE1)-tumor necrosis factor receptor associated factor 2 (TRAF2) signaling proteins were measured in high glucose and bupivacaine-treated cells. Cell viability was measured using a Cell Counting Kit-8 assay, cell apoptosis was assessed using flow cytometry, and protein expression was determined using western blot analyses. Compared with the control group, high glucose and bupivacaine significantly increased ATF4, CHOP and caspase-12 expression, increased apoptosis, and decreased p-IRE1, TRAF2, LC3-II/LC3-I and Beclin1 expression. Promoting autophagy with RAPA partly reversed the high glucose and bupivacaine-induced changes in p-PERK, CHOP, TRAF2, Beclin1, caspase-12 and apoptosis, while inhibiting autophagy with 3-MA further enhanced the changes in ATF4, CHOP, p-IRE1, TRAF2 and apoptosis. High glucose and bupivacaine induced cytotoxicity in SH-SY5Y cells, at least in part, through enhancing cell apoptosis and inhibiting autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways.
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spelling pubmed-66912382019-08-19 High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways Liu, Yongzhe Sun, Li Ma, Yaqun Wei, Biyu Gao, Minglong Shang, Lixin Mol Med Rep Articles Bupivacaine has previously been reported to induce neurotoxicity, which is further enhanced by high glucose levels. In the present study, the underlying molecular mechanisms via which bupivacaine induces cytotoxicity under high glucose conditions were investigated in cultured human SH-SY5Y cells. In order to identify the optimal concentrations of glucose and bupivacaine that induced cytotoxicity, SH-SY5Y cells were treated with 30–100 mM glucose and 0.5–1.0 mM bupivacaine. Based on the dose response experiments, 50 mM glucose and 0.5 mM bupivacaine was used in the present study. The effects that 3-MA (autophagy inhibitor) and rapamycin (RAPA; autophagy inducer) exerted on cell apoptosis, autophagy and the expression of protein kinase R-like endoplasmic reticulum kinase (PERK)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) and inositol-requiring enzyme 1 (IRE1)-tumor necrosis factor receptor associated factor 2 (TRAF2) signaling proteins were measured in high glucose and bupivacaine-treated cells. Cell viability was measured using a Cell Counting Kit-8 assay, cell apoptosis was assessed using flow cytometry, and protein expression was determined using western blot analyses. Compared with the control group, high glucose and bupivacaine significantly increased ATF4, CHOP and caspase-12 expression, increased apoptosis, and decreased p-IRE1, TRAF2, LC3-II/LC3-I and Beclin1 expression. Promoting autophagy with RAPA partly reversed the high glucose and bupivacaine-induced changes in p-PERK, CHOP, TRAF2, Beclin1, caspase-12 and apoptosis, while inhibiting autophagy with 3-MA further enhanced the changes in ATF4, CHOP, p-IRE1, TRAF2 and apoptosis. High glucose and bupivacaine induced cytotoxicity in SH-SY5Y cells, at least in part, through enhancing cell apoptosis and inhibiting autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways. D.A. Spandidos 2019-09 2019-07-25 /pmc/articles/PMC6691238/ /pubmed/31524237 http://dx.doi.org/10.3892/mmr.2019.10524 Text en Copyright: © Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Liu, Yongzhe
Sun, Li
Ma, Yaqun
Wei, Biyu
Gao, Minglong
Shang, Lixin
High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title_full High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title_fullStr High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title_full_unstemmed High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title_short High glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the PERK-ATF4-CHOP and IRE1-TRAF2 signaling pathways
title_sort high glucose and bupivacaine-induced cytotoxicity is mediated by enhanced apoptosis and impaired autophagy via the perk-atf4-chop and ire1-traf2 signaling pathways
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6691238/
https://www.ncbi.nlm.nih.gov/pubmed/31524237
http://dx.doi.org/10.3892/mmr.2019.10524
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