Resveratrol‑mediated activation of SIRT1 inhibits the PERK‑eIF2α‑ATF4 pathway and mitigates bupivacaine‑induced neurotoxicity in PC12 cells

Luo,Yunpeng; Hu,Na; Zhao,Yang; Lai,Jian; Luo,Xi; Liu,Jingchen

doi:10.3892/etm.2023.12132

Resveratrol‑mediated activation of SIRT1 inhibits the PERK‑eIF2α‑ATF4 pathway and mitigates bupivacaine‑induced neurotoxicity in PC12 cells

Authors:
- Yunpeng Luo
- Na Hu
- Yang Zhao
- Jian Lai
- Xi Luo
- Jingchen Liu
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Affiliations: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Radiology, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China
Published online on: July 24, 2023 https://doi.org/10.3892/etm.2023.12132
Article Number: 433
Copyright: © Luo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Endoplasmic reticulum (ER) stress and apoptosis play significant roles in the development of neurotoxicity caused by bupivacaine (BUP). By activating sirtuin 1 (SIRT1), resveratrol (RSV) can regulate various cellular processes associated with anti‑oxidative stress, anti‑apoptosis and anti‑inflammatory responses, thereby exerting neuroprotective effects. However, it remains unknown whether the activation of SIRT1 by RSV is able to attenuate BUP‑induced ER stress and apoptosis. Therefore, the present study aimed to explore the effect of RSV on BUP‑induced cytotoxicity in PC12 cells and the underlying mechanism. Cell Counting Kit‑8 assays, flow cytometry and inverted phase‑contrast microscopy were used to assess the viability, apoptosis rate and morphological changes of the cells, respectively. Western blotting and immunofluorescence staining were used to analyze the levels of SIRT1, the apoptosis‑related proteins Bax, Bcl‑2 and cleaved caspase‑3, the ER stress‑related proteins glucose‑regulated protein 78, caspase‑12 and CHOP, and the protein kinase RNA‑like ER kinase (PERK)‑eukaryotic translation initiation factor 2 α (eIF2α)‑activating transcription factor 4 (ATF4) pathway‑associated proteins phosphorylated (p)‑PERK, PERK, p‑eIF2α, eIF2α and ATF4. The results revealed that BUP induced cell apoptosis and decreased cell viability, accompanied by the downregulation of SIRT1. However, RSV restored SIRT1 protein expression, downregulated the expression of the pro‑apoptotic protein Bax, upregulated the expression of the anti‑apoptotic protein Bcl‑2, decreased the apoptosis rate of the cells and increased cell viability. Furthermore, the anti‑apoptotic effects exhibited by RSV were associated with inhibition of the PERK‑eIF2α‑ATF4 pathway of ER stress. However, the protective effect of RSV was significantly mitigated by the SIRT1 inhibitor EX527. These results indicate that the activation of SIRT1 by RSV alleviates BUP‑induced PC12 cell ER stress and apoptosis via regulation of the PERK‑eIF2α‑ATF4 pathway. These findings offer insights into the molecular mechanism underlying BUP‑induced apoptosis and suggest the potential of RSV as a therapeutic agent against the neurotoxicity caused by BU

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1	Chen X, Xu Z, Lin R and Liu Z: Persistent cauda equina syndrome after cesarean section under combined spinal-epidural anesthesia: A case report. J Clin Anesth. 27:520–523. 2015.PubMed/NCBI View Article : Google Scholar
2	Ji J, Yan X, Li Z, Lai Z and Liu J: Therapeutic effects of intrathecal versus intravenous monosialoganglioside against bupivacaine-induced spinal neurotoxicity in rats. Biomed Pharmacother. 69:311–316. 2015.PubMed/NCBI View Article : Google Scholar
3	Lai J, Ji JM, Chen MY, Luo YP, Yu Y, Zhou G, Wei LL, Huang LS and Liu JC: Melatonin ameliorates bupivacaine-induced spinal neurotoxicity in rats by suppressing neuronal NLRP3 inflammasome activation. Neurosci Lett. 772(136472)2022.PubMed/NCBI View Article : Google Scholar
4	Zhao Y, Luo Y, Liu Z, Chen Y, Wei L, Luo X, Zhou G, Lai J, Ji J, Lin Y and Liu J: Ferrostatin-1 ameliorates bupivacaine-induced spinal neurotoxicity in rats by inhibiting ferroptosis. Neurosci Lett. 809(137308)2023.PubMed/NCBI View Article : Google Scholar
5	Marciniak SJ, Chambers JE and Ron D: Pharmacological targeting of endoplasmic reticulum stress in disease. Nat Rev Drug Discov. 21:115–140. 2022.PubMed/NCBI View Article : Google Scholar
6	Ghemrawi R and Khair M: Endoplasmic reticulum stress and unfolded protein response in neurodegenerative diseases. Int J Mol Sci. 21(6127)2020.PubMed/NCBI View Article : Google Scholar
7	Hetz C, Zhang K and Kaufman RJ: Mechanisms, regulation and functions of the unfolded protein response. Nat Rev Mol Cell Biol. 21:421–438. 2020.PubMed/NCBI View Article : Google Scholar
8	Li L, Zhang QG, Lai LY, Wen XJ, Zheng T, Cheung CW, Zhou SQ and Xu SY: Neuroprotective effect of ginkgolide B on bupivacaine-induced apoptosis in SH-SY5Y cells. Oxid Med Cell Longev. 2013(159864)2013.PubMed/NCBI View Article : Google Scholar
9	Liang Y, Ji J, Lin Y, He Y and Liu J: The Ganglioside GM-1 inhibits bupivacaine-induced neurotoxicity in mouse neuroblastoma Neuro2a cells. Cell Biochem Funct. 34:455–462. 2016.PubMed/NCBI View Article : Google Scholar
10	Liu B, Ji J, Feng Q, Luo X, Yan X, Ni Y, He Y, Mao Z and Liu J: Monosialoganglioside protects against bupivacaine-induced neurotoxicity caused by endoplasmic reticulum stress in rats. Drug Des Devel Ther. 13:707–718. 2019.PubMed/NCBI View Article : Google Scholar
11	Liu H, Xu S, Wang C, Deng Y, Xu B, Yang T, Sun J and Liu W: The beneficial role of sirtuin 1 in preventive or therapeutic options of neurodegenerative diseases. Neuroscience. 504:79–92. 2022.PubMed/NCBI View Article : Google Scholar
12	Tang BL and Chua CE: SIRT1 and neuronal diseases. Mol Aspects Med. 29:187–200. 2008.PubMed/NCBI View Article : Google Scholar
13	Fangma Y, Wan H, Shao C, Jin L and He Y: Research progress on the role of sirtuin 1 in cerebral ischemia. Cell Mol Neurobiol. 43:1769–1783. 2023.PubMed/NCBI View Article : Google Scholar
14	Yu X, Zhang S, Zhao D, Zhang X, Xia C, Wang T, Zhang M, Liu T, Huang W and Wu B: SIRT1 inhibits apoptosis in in vivo and in vitro models of spinal cord injury via microRNA-494. Int J Mol Med. 43:1758–1768. 2019.PubMed/NCBI View Article : Google Scholar
15	Zheng LN, Guo FQ, Li ZS, Wang Z, Ma JH, Wang T, Wei JF and Zhang WW: Dexmedetomidine protects against lidocaine-induced neurotoxicity through SIRT1 downregulation-mediated activation of FOXO3a. Hum Exp Toxicol. 39:1213–1223. 2020.PubMed/NCBI View Article : Google Scholar
16	Wang X, Yuan B, Cheng B, Liu Y, Zhang B, Wang X, Lin X, Yang B and Gong G: Crocin alleviates myocardial ischemia/reperfusion-induced endoplasmic reticulum stress via regulation of miR-34a/Sirt1/Nrf2 pathway. Shock. 51:123–130. 2019.PubMed/NCBI View Article : Google Scholar
17	Li YP, Wang SL, Liu B, Tang L, Kuang RR, Wang XB, Zhao C, Song XD, Cao XM, Wu X, et al: Sulforaphane prevents rat cardiomyocytes from hypoxia/reoxygenation injury in vitro via activating SIRT1 and subsequently inhibiting ER stress. Acta Pharmacol Sin. 37:344–353. 2016.PubMed/NCBI View Article : Google Scholar
18	Zheng X, Xu F, Liang H, Cao H, Cai M, Xu W and Weng J: SIRT1/HSF1/HSP pathway is essential for exenatide-alleviated, lipid-induced hepatic endoplasmic reticulum stress. Hepatology. 66:809–824. 2017.PubMed/NCBI View Article : Google Scholar
19	He B, Zhang W, Qiao J, Peng Z and Chai X: Melatonin protects against COPD by attenuating apoptosis and endoplasmic reticulum stress via upregulating SIRT1 expression in rats. Can J Physiol Pharmacol. 97:386–391. 2019.PubMed/NCBI View Article : Google Scholar
20	Melhem H, Hansmannel F, Bressenot A, Battaglia-Hsu SF, Billioud V, Alberto JM, Gueant JL and Peyrin-Biroulet L: Methyl-deficient diet promotes colitis and SIRT1-mediated endoplasmic reticulum stress. Gut. 65:595–606. 2016.PubMed/NCBI View Article : Google Scholar
21	Kang X, Yang W, Wang R, Xie T, Li H, Feng D, Jin X, Sun H and Wu S: Sirtuin-1 (SIRT1) stimulates growth-plate chondrogenesis by attenuating the PERK-eIF-2α-CHOP pathway in the unfolded protein response. J Biol Chem. 293:8614–8625. 2018.PubMed/NCBI View Article : Google Scholar
22	Shaito A, Posadino AM, Younes N, Hasan H, Halabi S, Alhababi D, Al-Mohannadi A, Abdel-Rahman WM, Eid AH, Nasrallah GK and Pintus G: Potential adverse effects of resveratrol: A literature review. Int J Mol Sci. 21(2084)2020.PubMed/NCBI View Article : Google Scholar
23	Luo Y, Zhao Y, Lai J, Wei L, Zhou G, Yu Y and Liu J: Resveratrol suppresses bupivacaine-induced spinal neurotoxicity in rats by inhibiting endoplasmic reticulum stress via SIRT1 modulation. Biomed Res Int. 2023(1176232)2023.PubMed/NCBI View Article : Google Scholar
24	Fan X, Bian W, Liu M, Li J and Wang Y: MiRNA-429 alleviates ketamine-induced neurotoxicity through targeting BAG5. Environ Toxicol. 36:620–627. 2021.PubMed/NCBI View Article : Google Scholar
25	Hao R, Ge J, Song X, Li F, Sun-Waterhouse D and Li D: Cadmium induces ferroptosis and apoptosis by modulating miR-34a-5p/Sirt1axis in PC12 cells. Environ Toxicol. 37:41–51. 2022.PubMed/NCBI View Article : Google Scholar
26	Shayan M, Mehri S, Razavi BM and Hosseinzadeh H: Minocycline protects PC12 cells against cadmium-induced neurotoxicity by modulating apoptosis. Biol Trace Elem Res. 201:1946–1954. 2023.PubMed/NCBI View Article : Google Scholar
27	Tang XP, Guo XH, Geng D and Weng LJ: d-Limonene protects PC12 cells against corticosterone-induced neurotoxicity by activating the AMPK pathway. Environ Toxicol Pharmacol. 70(103192)2019.PubMed/NCBI View Article : Google Scholar
28	Zhang Y, He Y, Deng N, Chen Y, Huang J and Xie W: Protective effect of resveratrol against corticosterone-induced neurotoxicity in PC12 cells. Transl Neurosci. 10:235–240. 2019.PubMed/NCBI View Article : Google Scholar
29	Yang Z, Hu S, He Y and Ji L: LINC00665 rescues bupivacaine induced neurotoxicity in human neural cell of SH-SY5Y through has-miR-34a-5p. Brain Res Bull. 177:210–216. 2021.PubMed/NCBI View Article : Google Scholar
30	Jęśko H, Wencel P, Strosznajder RP and Strosznajder JB: Sirtuins and their roles in brain aging and neurodegenerative disorders. Neurochem Res. 42:876–890. 2017.PubMed/NCBI View Article : Google Scholar
31	Zhang Y, Li T, Pan M, Wang W, Huang W, Yuan Y, Xie Z, Chen Y, Peng J, Li X and Meng Y: SIRT1 prevents cigarette smoking-induced lung fibroblasts activation by regulating mitochondrial oxidative stress and lipid metabolism. J Transl Med. 20(222)2022.PubMed/NCBI View Article : Google Scholar
32	Ren Q, Hu Z, Jiang Y, Tan X, Botchway BOA, Amin N, Lin G, Geng Y and Fang M: SIRT1 protects against apoptosis by promoting autophagy in the oxygen glucose deprivation/reperfusion-induced injury. Front Neurol. 10(1289)2019.PubMed/NCBI View Article : Google Scholar
33	Jiao F and Gong Z: The beneficial roles of SIRT1 in neuroinflammation-related diseases. Oxid Med Cell Longev. 2020(6782872)2020.PubMed/NCBI View Article : Google Scholar
34	Chen H, Lin X, Yi X, Liu X, Yu R, Fan W, Ling Y, Liu Y and Xie W: SIRT1-mediated p53 deacetylation inhibits ferroptosis and alleviates heat stress-induced lung epithelial cells injury. Int J Hyperthermia. 39:977–986. 2022.PubMed/NCBI View Article : Google Scholar
35	Chen L, Li S, Zhu J, You A, Huang X, Yi X and Xue M: Mangiferin prevents myocardial infarction-induced apoptosis and heart failure in mice by activating the Sirt1/FoxO3a pathway. J Cell Mol Med. 25:2944–2955. 2021.PubMed/NCBI View Article : Google Scholar
36	Kaewmool C, Kongtawelert P, Phitak T, Pothacharoen P and Udomruk S: Protocatechuic acid inhibits inflammatory responses in LPS-activated BV2 microglia via regulating SIRT1/NF-κB pathway contributed to the suppression of microglial activation-induced PC12 cell apoptosis. J Neuroimmunol. 341(577164)2020.PubMed/NCBI View Article : Google Scholar
37	Mao H, Wang L, Xiong Y, Jiang G and Liu X: Fucoxanthin attenuates oxidative damage by activating the Sirt1/Nrf2/HO-1 signaling pathway to protect the kidney from ischemia-reperfusion injury. Oxid Med Cell Longev. 2022(7444430)2022.PubMed/NCBI View Article : Google Scholar
38	Wang F, Ma J, Wang J, Chen M, Xia H, Yao S and Zhang D: SIRT1 ameliorated septic associated-lung injury and macrophages apoptosis via inhibiting endoplasmic reticulum stress. Cell Signal. 97(110398)2022.PubMed/NCBI View Article : Google Scholar
39	Wang H, Dong X, Liu Z, Zhu S, Liu H, Fan W, Hu Y, Hu T, Yu Y, Li Y, et al: Resveratrol suppresses rotenone-induced neurotoxicity through activation of SIRT1/Akt1 signaling pathway. Anat Rec (Hoboken). 301:1115–1125. 2018.PubMed/NCBI View Article : Google Scholar
40	Tang X, Zhao Y, Zhou Z, Yan J, Zhou B, Chi X, Luo A and Li S: Resveratrol mitigates sevoflurane-induced neurotoxicity by the SIRT1-dependent regulation of BDNF expression in developing mice. Oxid Med Cell Longev. 2020(9018624)2020.PubMed/NCBI View Article : Google Scholar
41	Bai L, Liu R, Wang R, Xin Y, Wu Z, Ba Y, Zhang H, Cheng X, Zhou G and Huang H: Attenuation of Pb-induced Aβ generation and autophagic dysfunction via activation of SIRT1: Neuroprotective properties of resveratrol. Ecotoxicol Environ Saf. 222(112511)2021.PubMed/NCBI View Article : Google Scholar
42	Gomes BAQ, Silva JPB, Romeiro CFR, Dos Santos SM, Rodrigues CA, Gonçalves PR, Sakai JT, Mendes PFS, Varela ELP and Monteiro MC: Neuroprotective mechanisms of resveratrol in Alzheimer's disease: Role of SIRT1. Oxid Med Cell Longev. 2018(8152373)2018.PubMed/NCBI View Article : Google Scholar
43	Li L, Ye XP, Lu AZ, Zhou SQ, Liu H, Liu ZJ, Jiang S and Xu SY: Hyperglycemia magnifies bupivacaine-induced cell apoptosis triggered by mitochondria dysfunction and endoplasmic reticulum stress. J Neurosci Res. 91:786–798. 2013.PubMed/NCBI View Article : Google Scholar
44	Iurlaro R and Muñoz-Pinedo C: Cell death induced by endoplasmic reticulum stress. FEBS J. 283:2640–2652. 2016.PubMed/NCBI View Article : Google Scholar
45	Zhang X, Jiang L, Chen H, Wei S, Yao K, Sun X, Yang G, Jiang L, Zhang C, Wang N, et al: Resveratrol protected acrolein-induced ferroptosis and insulin secretion dysfunction via ER-stress-related PERK pathway in MIN6 cells. Toxicology. 465(153048)2022.PubMed/NCBI View Article : Google Scholar
46	Shati AA: Resveratrol protects against cadmium chloride-induced hippocampal neurotoxicity by inhibiting ER stress and GAAD 153 and activating sirtuin 1/AMPK/Akt. Environ Toxicol. 34:1340–1353. 2019.PubMed/NCBI View Article : Google Scholar
47	Zhu H, Li X, Qiao M, Sun X and Li G: Resveratrol alleviates inflammation and ER stress through SIRT1/NRF2 to delay ovarian aging in a short-lived fish. J Gerontol A Biol Sci Med Sci. 78:596–602. 2023.PubMed/NCBI View Article : Google Scholar
48	Chou X, Ding F, Zhang X, Ding X, Gao H and Wu Q: Sirtuin-1 ameliorates cadmium-induced endoplasmic reticulum stress and pyroptosis through XBP-1s deacetylation in human renal tubular epithelial cells. Arch Toxicol. 93:965–986. 2019.PubMed/NCBI View Article : Google Scholar
49	Zhang Y, He L, Tu M, Huang M, Chen Y, Pan D, Peng J and Shen X: The ameliorative effect of terpinen-4-ol on ER stress-induced vascular calcification depends on SIRT1-mediated regulation of PERK acetylation. Pharmacol Res. 170(105629)2021.PubMed/NCBI View Article : Google Scholar
50	Prola A, Pires Da Silva J, Guilbert A, Lecru L, Piquereau J, Ribeiro M, Mateo P, Gressette M, Fortin D, Boursier C, et al: SIRT1 protects the heart from ER stress-induced cell death through eIF2α deacetylation. Cell Death Differ. 24:343–356. 2017.PubMed/NCBI View Article : Google Scholar