Abstract
Morphine tolerance is an important factor in unsatisfactory analgesia. HADHA is a crucial enzyme in fatty acid β-oxidation. In this study, we investigated the potential significance of HADHA in a mechanism that might cause morphine tolerance related to functional changes in energy metabolism and further explored the effect of HADHA desuccinylation on morphine tolerance. Rats received daily intrathecal injections of 10 µg of morphine for a duration of 7 consecutive days, and pain thresholds were measured using the mechanical withdrawal threshold (MWT) and thermal tail flick latency (TFL) tests. µ-Opioid receptor (MOR), LC3-I/II, and P62 expression and HADHA succinylation were assessed. HADHA succinylation was analyzed by liquid chromatography–tandem mass spectrometry (LC‒MS/MS) and parallel reaction monitoring (PRM). Morphine influenced the LC3II/LC3I ratio and P62 expression level, which are crucial indicators of autophagy, and stimulated HADHA succinylation. Additionally, HADHA was selectively bound by the desuccinylase SIRT5, and SIRT5 overexpression decreased HADHA succinylation, reduced P62 expression, and alleviated morphine tolerance.
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Data availability
The datasets generated during and analyzed during the current study are available from the corresponding author on reasonable request.
References
Allouche S, Noble F, Marie N (2014) Opioid receptor desensitization: mechanisms and its link to tolerance. Front Pharmacol 5:280. https://doi.org/10.3389/fphar.2014.00280
Andrieux P, Chevillard C, Cunha-Neto E, Nunes JPS (2021) Mitochondria as a cellular hub in infection and inflammation. Int J Mol Sci 22(21):11338. https://doi.org/10.3390/ijms222111338
Antolak A, Bodzoń-Kułakowska A, Cetnarska E, Pietruszka M, Marszałek-Grabska M, Kotlińska J, Suder P (2017) Proteomic data in morphine addiction versus real protein activity: metabolic enzymes. J Cell Biochem 118(12):4323–4330. https://doi.org/10.1002/jcb.26085
Bodzon-Kulakowska A, Antolak A, Drabik A, Marszalek-Grabska M, Kotlińska J, Suder P (2017) Brain lipidomic changes after morphine, cocaine and amphetamine administration — DESI — MS imaging study. Biochim Biophys Acta (BBA) Mol Cell Biol Lipids 1862(7):686–691. https://doi.org/10.1016/j.bbalip.2017.04.003
Cecatto C, Hickmann FH, Rodrigues MD, Amaral AU, Wajner M (2015) Deregulation of mitochondrial functions provoked by long-chain fatty acid accumulating in long-chain 3-hydroxyacyl-CoA dehydrogenase and mitochondrial permeability transition deficiencies in rat heart-mitochondrial permeability transition pore opening as a potential contributing pathomechanism of cardiac alterations in these disorders. FEBS J 282(24):4714–4726. https://doi.org/10.1111/febs.13526
Du J, Zhou Y, Su X, Yu JJ, Khan S, Jiang H, Kim J, Woo J, Kim JH, Choi BH, He B, Chen W, Zhang S, Cerione RA, Auwerx J, Hao Q, Lin H (2011) Sirt5 is a NAD-dependent protein lysine demalonylase and desuccinylase. Science (New York, N.Y.) 334(6057):806–809. https://doi.org/10.1126/science.1207861
Foomani FH, Jarzembowski JA, Mostaghimi S, Mehrvar S, Kumar SN, Ranji M (2021) Optical metabolic imaging of mitochondrial dysfunction on HADH mutant newborn rat hearts. IEEE J Transl Eng Health Med 9:1800407. https://doi.org/10.1109/JTEHM.2021.3104966
Hirschey MD, Zhao Y (2015) Metabolic regulation by lysine malonylation, succinylation, and glutarylation. Mol Cell Proteomics MCP 14(9):2308–2315. https://doi.org/10.1074/mcp.R114.046664
Hu Z, Deng Y, Hu C, Deng P, Bu Q, Yan G, Zhou J, Shao X, Zhao J, Li Y, Zhu R, Xu Y, Zhao Y, Cen X (2012) 1H NMR-based metabonomic analysis of brain in rats of morphine dependence and withdrawal intervention. Behav Brain Res 231(1):11–19. https://doi.org/10.1016/j.bbr.2012.02.026
Johnston IN, Westbrook RF (2005) Inhibition of morphine analgesia by LPS: role of opioid and NMDA receptors and spinal glia. Behav Brain Res 156(1):75–83. https://doi.org/10.1016/j.bbr.2004.05.006
Kang W, Suzuki M, Saito T, Miyado K (2021) Emerging role of TCA cycle-related enzymes in human diseases. Int J Mol Sci 22(23):13057. https://doi.org/10.3390/ijms222313057
Kong H, Jiang CY, Hu L, Teng P, Zhang Y, Pan XX, Sun XD, Liu WT (2019) Morphine induces dysfunction of PINK1/Parkin-mediated mitophagy in spinal cord neurons implying involvement in antinociceptive tolerance. J Mol Cell Biol 11(12):1056–1068. https://doi.org/10.1093/jmcb/mjz002
Kumar S, Lombard DB (2018) Functions of the sirtuin deacylase SIRT5 in normal physiology and pathobiology. Crit Rev Biochem Mol Biol 53(3):311–334. https://doi.org/10.1080/10409238.2018.1458071
Lou G, Palikaras K, Lautrup S, Scheibye-Knudsen M, Tavernarakis N, Fang EF (2020) Mitophagy and neuroprotection. Trends Mol Med 26(1):8–20. https://doi.org/10.1016/j.molmed.2019.07.002
Lu W, Zhang R, Sheng W, Feng L, Xu P, Wang Y, Xie Y, Xu H, Wang G, Aa J (2021) Identification of morphine and heroin-treatment in mice using metabonomics. Metabolites 11(9):607. https://doi.org/10.3390/metabo11090607
Maeyashiki C, Oshima S, Otsubo K, Kobayashi M, Nibe Y, Matsuzawa Y, Onizawa M, Nemoto Y, Nagaishi T, Okamoto R, Tsuchiya K, Nakamura T, Watanabe M (2017) HADHA, the alpha subunit of the mitochondrial trifunctional protein, is involved in long-chain fatty acid-induced autophagy in intestinal epithelial cells. Biochem Biophys Res Commun 484(3):636–641. https://doi.org/10.1016/j.bbrc.2017.01.159
Mao J, Sung B, Ji RR, Lim G (2002) Chronic morphine induces downregulation of spinal glutamate transporters: implications in morphine tolerance and abnormal pain sensitivity. J Neurosci 22(18):8312–8323. https://doi.org/10.1523/JNEUROSCI.22-18-08312.2002
Martini L, Whistler JL (2007) The role of mu opioid receptor desensitization and endocytosis in morphine tolerance and dependence. Curr Opin Neurobiol 17(5):556–564. https://doi.org/10.1016/j.conb.2007.10.004
Raehal KM, Bohn LM (2014) β-arrestins: regulatory role and therapeutic potential in opioid and cannabinoid receptor-mediated analgesia. Handb Exp Pharmacol 219:427–443. https://doi.org/10.1007/978-3-642-41199-1_22
Ramshini E, Alaei H, Reisi P, Naghdi N, Afrozi H, Alaei S, Alehashem M, Eftekharvaghefi S (2019) Effect of intracerebroventricular injection of GABA receptors antagonists on morphine-induced changes in GABA and GLU transmission within the mPFC: an in vivo microdialysis study. Iran J Basic Med Sci 22(3):246–250. https://doi.org/10.22038/ijbms.2019.28478.6925
Rardin MJ, He W, Nishida Y, Newman JC, Carrico C, Danielson SR, Guo A, Gut P, Sahu AK, Li B, Uppala R, Fitch M, Riiff T, Zhu L, Zhou J, Mulhern D, Stevens RD, Ilkayeva OR, Newgard CB, Jacobson MP, Hellerstein M, Goetzman ES, Gibson BW, Verdin E (2013) SIRT5 regulates the mitochondrial lysine succinylome and metabolic networks. Cell Metab 18(6):920–933. https://doi.org/10.1016/j.cmet.2013.11.013
Sadakierska-Chudy A, Frankowska M, Filip M (2014) Mitoepigenetics and drug addiction. Pharmacol Ther 144(2):226–233. https://doi.org/10.1016/j.pharmthera.2014.06.002
Shibata M, Yoshimura K, Furuya N, Koike M, Ueno T, Komatsu M, Arai H, Tanaka K, Kominami E, Uchiyama Y (2009) The MAP1-LC3 conjugation system is involved in lipid droplet formation. Biochem Biophys Res Commun 382(2):419–423. https://doi.org/10.1016/j.bbrc.2009.03.039
Su LY, Liu Q, Jiao L, Yao YG (2021) Molecular mechanism of neuroprotective effect of melatonin on morphine addiction and analgesic tolerance: an update. Mol Neurobiol 58(9):4628–4638. https://doi.org/10.1007/s12035-021-02448-0
Tong Y, Guo D, Lin S, Liang J, Yang D, Ma C, Shao F, Li M, Yu Q, Jiang Y, Li L, Fang J, Yu R, Lu Z (2021) SUCLA2-coupled regulation of GLS succinylation and activity counteracts oxidative stress in tumor cells. Mol Cell 81(11):2303–2316. https://doi.org/10.1016/j.molcel.2021.04.002
Wan J, Ma J, Anand V, Ramakrishnan S, Roy S (2015) Morphine potentiates LPS-induced autophagy initiation but inhibits autophagosomal maturation through distinct TLR4-dependent and independent pathways. Acta Physiol (Oxford, England) 214(2):189–199. https://doi.org/10.1111/apha.12506
Paxinos G, Watson C (2013) The rat brain in stereotaxic coordinates seventh edition. Academic Press, New York
Weinert BT, Schölz C, Wagner SA, Iesmantavicius V, Su D, Daniel JA, Choudhary C (2013) Lysine succinylation is a frequently occurring modification in prokaryotes and eukaryotes and extensively overlaps with acetylation. Cell Rep 4(4):842–851. https://doi.org/10.1016/j.celrep.2013.07.024
Williams JT, Ingram SL, Henderson G, Chavkin C, von Zastrow M, Schulz S, Koch T, Evans CJ, Christie MJ (2013) Regulation of μ-opioid receptors: desensitization, phosphorylation, internalization, and tolerance. Pharmacol Rev 65(1):223–254. https://doi.org/10.1124/pr.112.005942
Xiao ZP, Lv T, Hou PP, Manaenko A, Liu Y, Jin Y, Gao L, Jia F, Tian Y, Li P, Zhang JH, Hu Q, Zhang X (2021) Sirtuin 5-mediated lysine desuccinylation protects mitochondrial metabolism following subarachnoid hemorrhage in mice. Stroke 52(12):4043–4053. https://doi.org/10.1161/STROKEAHA.121.034850
Yang Y, Gibson GE (2019) Succinylation links metabolism to protein functions. Neurochem Res 44(10):2346–2359. https://doi.org/10.1007/s11064-019-02780-x
Yang L, Guan G, Lei L, Lv Q, Liu S, Zhan X, Jiang Z, Gu X (2018) Palmitic acid induces human osteoblast-like Saos-2 cell apoptosis via endoplasmic reticulum stress and autophagy. Cell Stress Chaperones 23(6):1283–1294. https://doi.org/10.1007/s12192-018-0936-8
Yang YH, Wu SF, Zhu YP, Yang JT, Liu JF (2022) Global profiling of lysine succinylation in human lungs. Proteomics 22(17):e2100381. https://doi.org/10.1002/pmic.202100381
Zaitsu K, Miyawaki I, Bando K, Horie H, Shima N, Katagi M, Tatsuno M, Bamba T, Sato T, Ishii A, Tsuchihashi H, Suzuki K, Fukusaki E (2014) Metabolic profiling of urine and blood plasma in rat models of drug addiction on the basis of morphine, methamphetamine, and cocaine-induced conditioned place preference. Anal Bioanal Chem 406(5):1339–1354. https://doi.org/10.1007/s00216-013-7234-1
Zorov DB, Juhaszova M, Sollott SJ (2014) Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev 94(3):909–950. https://doi.org/10.1152/physrev.00026.2013
Acknowledgements
The authors acknowledge Guiyang Beilong Biotechnology Co., Ltd., for providing the necessary reagents.
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This study was funded by the Science and Technology Program of Guizhou Provincial Health Commission (gzwkj2023-395) and Guizhou Province Science and Technology Plan Project (grant no.: Qianke He Foundation‐ZK[2021]General 508).
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WL provided the overall design of the experiment. SHL and YXH both performed the experiment and analyzed the data. YC and YYF provided helpful comments. SHL completed the manuscript with the help of WL. All authors have read and approved the manuscript. The authors declare that all data were generated in-house and that no paper mill was used.
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Yuanxin Huang and Sihui Lu share first authorship.
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Huang, Y., Lu, S., Chen, Y. et al. Morphine induces HADHA succinylation, while HADHA desuccinylation alleviates morphine tolerance by influencing autophagy. Naunyn-Schmiedeberg's Arch Pharmacol 397, 1589–1600 (2024). https://doi.org/10.1007/s00210-023-02697-x
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DOI: https://doi.org/10.1007/s00210-023-02697-x