Crocin’s effect on phenotype switching of J774A.1 macrophages depends on their polarization state during exposure

Document Type : Original Article

Authors

1 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Cardiovascular Diseases Research Center, Birjand University of Medical Sciences, Birjand, Iran

3 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

5 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Macrophages exhibit versatile phenotypes, with M1 macrophages releasing inflammatory cytokines and possessing microbicidal activities, while M2 macrophages release anti-inflammatory cytokines and contribute to tissue repair. The M1/M2 imbalance plays a significant role in various pathological processes. Crocin, known for its antioxidant properties and ability to eliminate free radicals, has been investigated for its potential anti-inflammatory effects. We examined the effect of the primary activation state of macrophages on their phenotype switching when exposed to crocin.
Materials and Methods: The crocin impact on macrophage viability was evaluated by MTT. TNF-α, IL-6, and IL-10 secretion, as well as Nos2/Arg1 ratio, were measured in cells treated with crocin or LPS+IFN-γ (M1 inducers), in cells concurrently treated with crocin and LPS+IFN-γ or in cells pretreated with crocin before M1 induction.
Results: Crocin did not show any toxicity at the concentration of 500 µM or lower. When uncommitted macrophages were exposed to crocin (25-100 µM), it elevated certain M1 activity indicators, including Nos2/Arg1 ratio and TNF-α secretion, but not IL-6. Crocin in concurrent treatment with LPS+IFN-γ prevented the increase in M1 indicators, Nos2/Arg1 ratio, and TNF-α secretion. However, pretreatment of cells with crocin before the addition of LPS+IFN-γ did not reverse M1 induction in macrophages; instead, it further increased the Nos2/Arg1 ratio and TNF-α secretion. IL-10 was not detectable in any of the experimental groups.
Conclusion: It appears that the modulatory effects of crocin on macrophage M1/M2 phenotype switching partly depend on the presence or absence of inflammatory mediators and, accordingly, the initial state of macrophage commitment.

Keywords

Main Subjects

References

1. Davies LC, Jenkins SJ, Allen JE, Taylor PR. Tissue-resident macrophages. Nat Immunol 2013; 14:986-995.
2. Martinez FO, Gordon S. The M1 and M2 paradigm of macrophage activation: Time for reassessment. F1000Prime Rep 2014; 6:13-25.
3. Aras S, Zaidi MR. TAMeless traitors: Macrophages in cancer progression and metastasis. Br J cancer 2017; 117:1583-1591.
4. Mills C. M1 and M2 macrophages: Oracles of health and disease. Crit Rev Immunol 2012; 32:463-488.
5. Jayasingam SD, Citartan M, Thang TH, Mat Zin AA, Ang KC, Ch’ng ES. Evaluating the polarization of tumor-associated macrophages into M1 and M2 phenotypes in human cancer tissue: Technicalities and challenges in routine clinical practice. Front Oncol 2020; 9:1512-1520.
6. Kieler M, Hofmann M, Schabbauer G. More than just protein building blocks: How amino acids and related metabolic pathways fuel macrophage polarization. FEBS J 2021; 288:3694-3714.
7. Röszer T. Adipose tissue immunometabolism and apoptotic cell clearance. Cells 2021; 10:2288-2303.
8. Askari VR, Shafiee-Nick R. The protective effects of β-caryophyllene on LPS-induced primary microglia M1/M2 imbalance: A mechanistic evaluation. Life Sci 2019; 219:40-73.
9. Liu L-L, Li J-M, Su W-J, Wang B, Jiang C-L. Sex differences in depressive-like behaviour may relate to imbalance of microglia activation in the hippocampus. Brain Behav Immun 2019; 81:188-197.
10. Rahmanian-Devin P, Rakhshandeh H, Baradaran Rahimi V, Sanei-Far Z, Hasanpour M, Memarzia A, et al. Intraperitoneal Lavage with Crocus sativus Prevents Postoperative-Induced Peritoneal Adhesion in a Rat Model: Evidence from Animal and Cellular Studies. Oxid Med Cell Longev 2021;2021:5945101-5945122.
11. Yu T, Zhao L, Huang X, Ma C, Wang Y, Zhang J, et al. Enhanced activity of the macrophage M1/M2 phenotypes and phenotypic switch to M1 in periodontal infection. J Periodontol 2016; 87:1092-1102.
12. Pellegrini C, Fornai M, Colucci R, Tirotta E, Blandini F, Levandis G, et al. Alteration of colonic excitatory tachykininergic motility and enteric inflammation following dopaminergic nigrostriatal neurodegeneration. J Neuroinflammation 2016; 13:1-13.
13. El-Gayar S, Thüring-Nahler hr Pfeilschifter J, Röllinghoff M, Bogdan C. Translational control of inducible nitric oxide synthase by IL-13 and arginine availability in inflammatory macrophages. J Immunol 2003; 171:4561-4568.
14. Mills CD. Anatomy of a discovery: M1 and m2 macrophages. Front Immunol 2015; 6:212-223.
15. Mantovani A, Garlanda C, Locati M. Macrophage diversity and polarization in atherosclerosis: A question of balance. Arterioscler Thromb Vasc Biol 2009; 29:1419-1423.
16. Watzl B. Anti-inflammatory effects of plant-based foods and of their constituents. Int J Vitam Nutr Res 2008; 78:293-298.
17. Voutilainen S, Nurmi T, Mursu J, Rissanen TH. Carotenoids and cardiovascular health. Am J Clin Nutr 2006; 83:1265-1271.
18. Alavizadeh SH, Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: A comprehensive review. Food Chem Toxicol 2014; 64:65-80.
19. Nam KN, Park Y-M, Jung H-J, Lee JY, Min BD, Park S-U, et al. Anti-inflammatory effects of crocin and crocetin in rat brain microglial cells. Eur J Pharmacol 2010; 648:110-116.
20. Funes SC, Rios M, Escobar‐Vera J, Kalergis AM. Implications of macrophage polarization in autoimmunity. Immunology 2018; 154:186-195.
21. Bhutia YD, Babu E, Ganapathy V. Interferon-γ induces a tryptophan-selective amino acid transporter in human colonic epithelial cells and mouse dendritic cells. Biochim Biophys Acta 2015; 1848:453-462.
22. Choi J-W, Kwon M-J, Kim I-H, Kim Y-M, Lee MK, Nam T-J. Pyropia yezoensis glycoprotein promotes the M1 to M2 macrophage phenotypic switch via the STAT3 and STAT6 transcription factors. Int J Mol Med 2016; 38:666-674.
23. Ye S, Xu hr Jin J, Yang M, Wang C, Yu Y, et al. The E3 ubiquitin ligase neuregulin receptor degradation protein 1 (Nrdp1) promotes M2 macrophage polarization by ubiquitinating and activating transcription factor CCAAT/enhancer-binding Protein β (C/EBPβ). J Biol Chem 2012; 287:26740-26748.
24. Zeinali M, Zirak MR, Rezaee SA, Karimi G, Hosseinzadeh H. Immunoregulatory and anti-inflammatory properties of Crocus sativus (Saffron) and its main active constituents: A review. Iran J Basic Med Sci 2019; 22:334-344.
25. Hofkens W, Schelbergen R, Storm G, van den Berg WB, van Lent PL. Liposomal targeting of prednisolone phosphate to synovial lining macrophages during experimental arthritis inhibits M1 activation but does not favor M2 differentiation. PloS One 2013; 8:e54016-e54026.
26. Bakshi HA, Hakkim FL, Sam S. Molecular mechanism of crocin induced caspase mediated MCF-7 cell death: In vivo toxicity profiling and ex vivo macrophage activation. Asian Pac J Cancer Prev 2016; 17:1499-1506.
27. Xi L, Qian Z. Pharmacological properties of crocetin and crocin (digentiobiosyl ester of crocetin) from saffron. Nat Prod Commun 2006; 1:65-75.
28. Ghiasian M, Khamisabadi F, Kheiripour N, Karami M, Haddadi R, Ghaleiha A, et al. Effects of crocin in reducing DNA damage, inflammation, and oxidative stress in multiple sclerosis patients: A double‐blind, randomized, and placebo‐controlled trial. J Biochem Mol Toxicol 2019; 33:e22410.
29. Abdi hr Aganj Z, Hosseinzadeh H, Mosaffa F. Crocin restores the balance of Th1/Th2 immune cell response in ConA-treated human lymphocytes. Pharmacol Rep 2022;74:513-522.
30. Oh W-J, Jung U, Eom H-S, Shin H-J, Park H-R. Inhibition of lipopolysaccharide-induced proinflammatory responses by Buddleja officinalis extract in BV-2 microglial cells via negative regulation of NF-κB and ERK1/2 signaling. Molecules 2013; 18:9195-9206.
31. Zhang Y, Choksi S, Chen K, Pobezinskaya Y, Linnoila I, Liu Z-G. ROS play a critical role in the differentiation of alternatively activated macrophages and the occurrence of tumor-associated macrophages. Cell Res 2013; 23:898-914.
32. Mohammadi E, Mehri S, Bostan HB, Hosseinzadeh H. Protective effect of crocin against d-galactose-induced aging in mice. Avicenna J Phytomed 2018; 8:14-23.
33. Omidkhoda SF, Mehri S, Heidari S, Hosseinzadeh H. Protective effects of crocin against hepatic damages in D-galactose aging model in rats. Iran J Pharm Res 2020; 19:440-450.
34. Shen Z. Crocin inhibits the melanoma invasion and metastasis by regulating the polarization phenotype of macrophage. Open Access Library Journal 2019; 6:1-11.
35. Piao M, Cao hr He N, Yang B, Dong W, Xu M, et al. Berberine inhibits intestinal polyps growth in Apc (min/+) mice via regulation of macrophage polarization. Evid Based Complement Alternat Med 2016;2016:5137505-5137512.
36. Zhang Y, Qiu Z, Qiu Y, Su T, Qu P, Jia A. Functional regulation of ginsenosides on myeloid immunosuppressive cells in the tumor microenvironment. Integr Cancer Ther 2019; 18:1534735419886655-1534735419886663.
37. Saw CLL, Yang AY, Cheng DC, Boyanapalli SS-S, Su Z-Y, Khor TO, et al. Pharmacodynamics of ginsenosides: antioxidant activities, activation of Nrf2, and potential synergistic effects of combinations. Chem Res Toxicol 2012; 25:1574-1580.
38. Behrouz V, Sohrab G, Hedayati M, Sedaghat M. Inflammatory markers response to crocin supplementation in patients with type 2 diabetes mellitus: A randomized controlled trial. Phytother Res 2021; 35:4022-4031.
39. Behrouz V, Dastkhosh A, Hedayati M, Sedaghat M, Sharafkhah M, Sohrab G. The effect of crocin supplementation on glycemic control, insulin resistance and active AMPK levels in patients with type 2 diabetes: A pilot study. Diabetol Metab Syndr 2020; 12:1-9.
40. Saberi‐Karimian M, Safarian‐Bana H, Mohammadzadeh E, Kazemi T, Mansoori A, Ghazizadeh H, et al. A pilot study of the effects of crocin on high‐density lipoprotein cholesterol uptake capacity in patients with metabolic syndrome: A randomized clinical trial. Biofactors 2021; 47:1032-1041.
41. Kermani T, Kazemi T, Molki S, Ilkhani K, Sharifzadeh G, Rajabi O. The efficacy of crocin of saffron (Crocus sativus L.) on the components of metabolic syndrome: A randomized controlled clinical trial. J Res Pharm Pract 2017; 6:228-232.
42. Dastkhosh A, Behrouz V, Sohrab G, Sedaghat M. Effects of crocin supplementation on lipid profile and oxidative stress in patients with type 2 diabetes: A randomized clinical trial.  2022.
43. Yao Y, Xu X-H, Jin L. Macrophage polarization in physiological and pathological pregnancy. Front Immunol 2019; 10:792-804.
Iranian Journal of Basic Medical Sciences
Volume 26, Issue 12
December 2023
Pages 1431-1437

Files

Share

How to cite

Statistics