Morphine enhances LPS‑induced macrophage apoptosis through a PPARγ‑dependent mechanism

Lin,Mingying; Deng,Keqiong; Li,Ya; Wan,Jing

doi:10.3892/etm.2021.10146

Morphine enhances LPS‑induced macrophage apoptosis through a PPARγ‑dependent mechanism

Authors:
- Mingying Lin
- Keqiong Deng
- Ya Li
- Jing Wan
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Affiliations: Department of Cardiology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, P.R. China
Published online on: May 3, 2021 https://doi.org/10.3892/etm.2021.10146
Article Number: 714
Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Morphine has been widely used for the treatment of pain and extensive studies have revealed a regulatory role for morphine in cell apoptosis. However, the molecular mechanisms underlying morphine‑mediated apoptosis remain to be fully elucidated. The present study aimed to investigate the effects of morphine on lipopolysaccharide (LPS)‑induced bone marrow‑derived macrophage (BMDM) apoptosis and to determine the role of the peroxisome proliferator‑activated receptor (PPAR)γ signaling pathway in this process. BMDMs were isolated from BALB/c mice and stimulated with LPS. Hoechst 33342 staining and flow cytometric analysis were performed to evaluate the effects of morphine on LPS‑induced apoptosis of BMDMs. Caspase activity assays were used to determine the involvement of the apoptosis pathway. The expression levels of caspase‑3, caspase‑8, caspase‑9 and PPARγ were analyzed using western blotting. Finally, GW9662, a specific PPARγ antagonist, was used to determine whether the regulatory effects of morphine on LPS‑induced BMDM apoptosis were PPARγ‑dependent. The results of the present study revealed that morphine increased the apoptosis of LPS‑stimulated BMDMs. Morphine upregulated the expression levels and activity of caspase‑3 in LPS‑stimulated BMDMs, but downregulated the expression levels and activity of caspase‑8. Morphine treatment also upregulated LPS‑induced PPARγ expression levels in BMDMs. Finally, the stimulatory effects of morphine on LPS‑induced apoptosis and caspase‑3/9 activation were markedly reduced by GW9662. In conclusion, the findings of the present study indicated that morphine significantly promoted LPS‑induced BMDM apoptosis and caspase‑3/9 activation. These results suggested that the intrinsic pathway of apoptosis may be involved in the proapoptotic effects of morphine on LPS‑stimulated BMDMs, which may be dependent, at least partially, on PPARγ activation.

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1	Puig S and Gutstein HB: Opioids: Keeping the good, eliminating the bad. Nat Med. 23:272–273. 2017.PubMed/NCBI View Article : Google Scholar
2	Tuerxun H and Cui J: The dual effect of morphine on tumor development. Clin Transl Oncol. 21:695–701. 2019.PubMed/NCBI View Article : Google Scholar
3	Plein LM and Rittner HL: Opioids and the immune system-friend or foe. Br J Pharmacol. 175:2717–2725. 2018.PubMed/NCBI View Article : Google Scholar
4	Bajic D, Commons KG and Soriano SG: Morphine-enhanced apoptosis in selective brain regions of neonatal rats. Int J Dev Neurosci. 31:258–266. 2013.PubMed/NCBI View Article : Google Scholar
5	Li Y, Li H, Zhang Y, Sun X, Hanley GA, LeSage G, Zhang Y, Sun S, Peng Y and Yin D: Toll-like receptor 2 is required for opioids-induced neuronal apoptosis. Biochem Bioph Res Commun. 391:426–430. 2010.PubMed/NCBI View Article : Google Scholar
6	Osmanloglu O, Yldirim MK, Akyuva Y, Yildizhan K and Naziroglu M: Morphine induces apoptosis, inflammation, and mitochondrial oxidative stress via activation of TRPM2 channel and nitric oxide signaling pathways in the hippocampus. Mol Neurobiol. 57:3376–3389. 2020.PubMed/NCBI View Article : Google Scholar
7	Weng HL and Wang MJ: Effects of microRNA-338-3p on morphine-induced apoptosis and its underlying mechanisms. Mol Med Rep. 14:2085–2092. 2016.PubMed/NCBI View Article : Google Scholar
8	Singh R, Letai A and Sarosiek K: Regulation of apoptosis in health and disease: The balancing act of BCL-2 family proteins. Nat Rev Mol Cell Biol. 20:175–193. 2019.PubMed/NCBI View Article : Google Scholar
9	Gonzalez L and Trigatti BL: Macrophage apoptosis and necrotic core development in atherosclerosis: A rapidly advancing field with clinical relevance to imaging and therapy. Can J Cardiol. 33:303–312. 2017.PubMed/NCBI View Article : Google Scholar
10	Davis TME, Peters KE and Lipscombe R: Apoptosis inhibitor of macrophage and diabetic kidney disease. Cell Mol Immunol. 16(521)2019.PubMed/NCBI View Article : Google Scholar
11	Li Z and Weinman SA: Regulation of hepatic inflammation via macrophage cell death. Semin Liver Dis. 38:340–350. 2018.PubMed/NCBI View Article : Google Scholar
12	George L, Ramasamy T, Sirajudeen KNS and Manickam V: LPS-induced apoptosis is partially mediated by hydrogen sulphide in RAW 264.7 murine macrophages. Immunol Invest. 48:451–465. 2019.PubMed/NCBI View Article : Google Scholar
13	Xaus J, Comalada M, Valledor AF, Lloberas J, López-Soriano F, Argilés JM, Bogdan C and Celada A: LPS induces apoptosis in macrophages mostly through the autocrine production of TNF-alpha. Blood. 95:3823–3831. 2000.PubMed/NCBI
14	Du P, Li SJ, Ojcius DM, Li KX, Hu WL, Lin X, Sun AH and Yan J: A novel Fas-binding outer membrane protein and lipopolysaccharide of Leptospira interrogans induce macrophage apoptosis through the Fas/FasL-caspase-8/-3 pathway. Emerg Microbes Infec. 7(135)2018.PubMed/NCBI View Article : Google Scholar
15	Bhat RS, Bhaskaran M, Mongia A, Hitosugi N and Singhal PC: Morphine-induced macrophage apoptosis: Oxidative stress and strategies for modulation. J Leukocyte Biol. 75:1131–1138. 2004.PubMed/NCBI View Article : Google Scholar
16	Ahmadian M, Suh JM, Hah N, Liddle C, Atkins AR, Downes M and Evans RM: PPARγ signaling and metabolism: The good, the bad and the future. Nat Med. 19:557–566. 2013.PubMed/NCBI View Article : Google Scholar
17	Wang SB, Dougherty EJ and Danner RL: PPARγ signaling and emerging opportunities for improved therapeutics. Pharmacol Res. 111:76–85. 2016.PubMed/NCBI View Article : Google Scholar
18	Mahmood DFD, Jguirim-Souissi I, Khadija EH, Blondeau N, Diderot V, Amrani S, Slimane MN, Syrovets T, Simmet T and Rouis M: Peroxisome proliferator-activated receptor gamma induces apoptosis and inhibits autophagy of human monocyte-derived macrophages via induction of cathepsin L: Potential role in atherosclerosis. J Biol Chem. 286:28858–28866. 2011.PubMed/NCBI View Article : Google Scholar
19	Billiet L, Furman C, Larigauderie G, Copin C, Page S, Fruchart JC, Brand K and Rouis M: Enhanced VDUP-1 gene expression by PPARgamma agonist induces apoptosis in human macrophage. J Cell Physiol. 214:183–191. 2008.PubMed/NCBI View Article : Google Scholar
20	Chinetti G, Griglio S, Antonucci M, Torra IP, Delerive P, Majd Z, Fruchart JC, Chapman J, Najib J and Staels B: Activation of proliferator-activated receptors alpha and gamma induces apoptosis of human monocyte-derived macrophages. J Biol Chem. 273:25573–25580. 1998.PubMed/NCBI View Article : Google Scholar
21	de Guglielmo G, Kallupi M, Scuppa G, Stopponi S, Demopulos G, Gaitanaris G and Ciccocioppo R: Analgesic tolerance to morphine is regulated by PPARγ. Br J Pharmacol. 171:5407–5416. 2014.PubMed/NCBI View Article : Google Scholar
22	Weischenfeldt J and Porse B: Bone marrow-derived macrophages (BMM): Isolation and applications. CSH Protoc 2008: pdb.prot5080, 2008.
23	Wan J, Xiao Z, Chao S, Xiong S, Gan X, Qiu X, Xu C, Ma Y and Tu X: Pioglitazone modulates the proliferation and apoptosis of vascular smooth muscle cells via peroxisome proliferators-activated receptor-gamma. Diabetol Metab Syndr. 6(101)2014.PubMed/NCBI View Article : Google Scholar
24	Pazár B, Ea HK, Narayan S, Kolly L, Bagnoud N, Chobaz V, Roger T, Lioté F, So A and Busso N: Basic calcium phosphate crystals induce monocyte/Macrophage IL-1β secretion through the NLRP3 inflammasome in vitro. J Immunol. 186:2495–2502. 2011.PubMed/NCBI View Article : Google Scholar
25	Li CG, Yan L, Mai FY, Shi ZJ, Xu LH, Jing YY, Zha QB, Ouyang DY and He XH: Baicalin inhibits NOD-like receptor family, pyrin containing domain 3 inflammasome activation in murine macrophages by augmenting protein kinase a signaling. Front Immunol. 8(1409)2017.PubMed/NCBI View Article : Google Scholar
26	Guo S, Sun X, Cheng J, Xu H, Dan J, Shen J, Zhou Q, Zhang Y, Meng L, Cao W and Tian Y: Apoptosis of THP-1 macrophages induced by protoporphyrin IX-mediated sonodynamic therapy. Int J Nanomedicine. 8:2239–2246. 2013.PubMed/NCBI View Article : Google Scholar
27	Fulda S and Debatin KM: Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 25:4798–4811. 2006.PubMed/NCBI View Article : Google Scholar
28	Brentnall M, Rodriguez-Menocal L, De Guevara RL, Cepero E and Boise LH: Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol. 14(32)2013.PubMed/NCBI View Article : Google Scholar
29	Kurokawa M and Kornbluth S: Caspases and kinases in a death grip. Cell. 138:838–854. 2009.PubMed/NCBI View Article : Google Scholar
30	Yenikomshian HA, Curtis EE, Carrougher GJ, Qiu Q, Gibran NS and Mandell SP: Outpatient opioid use of burn patients: A retrospective review. Burns. 45:1737–1742. 2019.PubMed/NCBI View Article : Google Scholar
31	Maher DP, Walia D and Heller NM: Morphine decreases the function of primary human natural killer cells by both TLR4 and opioid receptor signaling. Brain Behavior Immunity. 83:298–302. 2020.PubMed/NCBI View Article : Google Scholar
32	Kroemer G and Martin SJ: Caspase-independent cell death. Nat Med. 11:725–730. 2005.PubMed/NCBI View Article : Google Scholar
33	Shapouri-Moghaddam A, Mohammadian S, Vazini H, Taghadosi M, Esmaeili SA, Mardani F, Seifi B, Mohammadi A, Afshari JT and Sahebkar A: Macrophage plasticity, polarization, and function in health and disease. J Cell Physiol. 233:6425–6440. 2018.PubMed/NCBI View Article : Google Scholar
34	Javadi S, Ejtemaeimehr S, Keyvanfar HR, Moghaddas P, Aminian A, Rajabzadeh A, Mani AR and Dehpour AR: Pioglitazone potentiates development of morphine-dependence in mice: Possible role of NO/cGMP pathway. Brain Res. 1510:22–37. 2013.PubMed/NCBI View Article : Google Scholar
35	Zhao L, Zhu Y, Wang D, Chen M, Gao P, Xiao W, Rao G, Wang X, Jin H, Xu L, et al: Morphine induces Beclin 1-and ATG5-dependent autophagy in human neuroblastoma SH-SY5Y cells and in the rat hippocampus. Autophagy. 6:386–394. 2010.PubMed/NCBI View Article : Google Scholar
36	Koob GF: Neurobiology of opioid addiction: Opponent process, hyperkatifeia, and negative reinforcement. Biol Psychiat. 87:44–53. 2020.PubMed/NCBI View Article : Google Scholar
37	Task Force on the management of ST-segment elevation acute myocardial infarction of the European Society of Cardiology (ESC). Steg PG, James SK, Atar D, Badano LP, Blömstrom-Lundqvist C, Borger MA, Di Mario C, Dickstein K, Ducrocq G, et al: ESC guidelines for the management of acute myocardial infarction in patients presenting with ST-segment elevation. Eur Heart J. 33:2569–2619. 2012.PubMed/NCBI View Article : Google Scholar
38	O'Gara PT: 2013 ACCF/AHA guideline for the management of ST-elevation myocardial infarction: A report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines (vol 127, pp e362, 2013). Circulation. 128:e362–e425. 2013.PubMed/NCBI View Article : Google Scholar
39	Murphy GS, Szokol JW, Marymont JH, Avram MJ and Vender JS: Opioids and cardioprotection: The impact of morphine and fentanyl on recovery of ventricular function after cardiopulmonary bypass. J Cardiothor Vasc Anesth. 20:493–502. 2006.PubMed/NCBI View Article : Google Scholar
40	Tanaka K, Kersten JR and Riess ML: Opioid-induced cardioprotection. Curr Pharm Design. 20:5696–5705. 2014.PubMed/NCBI View Article : Google Scholar
41	Rentoukas I, Giannopoulos G, Kaoukis A, Kossyvakis C, Raisakis K, Driva M, Panagopoulou V, Tsarouchas K, Vavetsi S, Pyrgakis V and Deftereos S: Cardioprotective role of remote ischemic periconditioning in primary percutaneous coronary intervention enhancement by opioid action. JACC Cardiovasc Interv. 3:49–55. 2010.PubMed/NCBI View Article : Google Scholar
42	Wang Y, Wang L, Li JH, Zhao HW and Zhang FZ: Morphine alleviates myocardial ischemia/reperfusion injury in rats by inhibiting TLR4/NF-κB signaling pathway. Eur Rev Med Pharmaco. 23:8616–8624. 2019.PubMed/NCBI View Article : Google Scholar
43	Chen ZL, Li TZ and Zhang BX: Morphine postconditioning protects against reperfusion injury in the isolated rat hearts. J Surg Res. 145:287–294. 2008.PubMed/NCBI View Article : Google Scholar
44	Oppi S, Nusser-Stein S, Blyszczuk P, Wang X, Jomard A, Marzolla V, Yang K, Velagapudi S, Ward LJ, Yuan XM, et al: Macrophage NCOR1 protects from atherosclerosis by repressing a pro-atherogenic PPARγ signature. Eur Heart J. 41:995–1005. 2020.PubMed/NCBI View Article : Google Scholar
45	Subramanian M, Thorp E and Tabas I: Identification of a non-growth factor role for GM-CSF in advanced atherosclerosis promotion of macrophage apoptosis and plaque necrosis through IL-23 signaling. Circ Res. 116:e13–e24. 2015.PubMed/NCBI View Article : Google Scholar
46	Linton MF, Babaev VR, Huang JS, Linton EF, Tao H and Yancey PG: Macrophage apoptosis and efferocytosis in the pathogenesis of atherosclerosis. Circ J. 80:2259–2268. 2016.PubMed/NCBI View Article : Google Scholar