Anti-leishmanial, immunomodulatory and anti-oxidative activity of quercetin against cutaneous leishmaniasis caused by Leishmania major

Abstract

Objective: To evaluate the in vitro and in vivo efficacy of quercetin and its immunomodulatory and anti-oxidative activity against Leishmania major ( L. major). Methods: L. major promastigotes and amastigotes were incubated with different concentrations of quercetin to estimate EC₅₀. For in vivo study, the base of tails of mice was infected with L. major. After developing ulcers in the inoculation site, mice were treated with 50 mg/kg quercetin orally for 28 consecutive days. The wound-healing potential of quercetin was evaluated by histopathological analysis of tissue sections stained by hematoxylin and eosin as well as Masson's trichrome. In addition, the levels of tumor necrosis factor-α, interleukin-6, malondialdehyde, and adiponectin, the ferric reducing ability of plasma, as well as superoxide dismutase and glutathione peroxidase activities were measured. Results: The EC₅₀ values of quercetin against L. major promastigotes and intracellular amastigotes were 0.27 and 0.85 µM, respectively. Histopathological analysis showed that fewer inflammatory cells, more fibroblasts, and more collagen deposition were observed in tissue sections of quercetin-treated mice. In addition, treatment with quercetin markedly increased glutathione peroxidase activity, the ferric reducing ability of plasma and adiponectin levels while decreasing malondialdehyde, interleukin-6, and tumor necrosis factor-α levels. Conclusions: Quercetin shows anti-leishmanial activity, immunomodulatory, anti-oxidative, and anti-inflammatory effects. Therefore, it may be further explored as an effective drug in treating leishmaniasis.

Keywords

Leishmania major / Cutaneous leishmaniosis / Wound healing / Quercetin / Immunomodulator

Acknowledgments

The authors would like to thank the authorities of the Veterinary School, Shiraz University, for their kind cooperation. We also thank the INSF (grant number 96006039) for its financial support.

Conflict of interest statement

The authors declare there is no conflict of interest.

Funding

The study was financially supported by INSF (grant number 96006039).

Authors’ contributions

SB and AO conceived and designed the study. EB and SB analyzed the data. EB and SB interpreted the data. SB and EB drafted the manuscript. SB, EB, and AO critically revised the manuscript for intellectual content. All authors contributed to the article and approved the submitted version.

References

[1] Mittra B, Saha A, Chowdhury AR, Pal C, Mandal S, Mukhopadhyay S, et al. Luteolin, an abundant dietary component is a potent anti-leishmanial agent that acts by inducing topoisomerase II-mediated kinetoplast DNA cleavage leading to apoptosis. Mol Med 2000; 6(6): 527.

[2] Akbari M, Oryan A, Hatam G. Application of nanotechnology in treatment of leishmaniosis: A review. Acta Trop 2017; 172: 86-90.

[3] Daneshbod Y, Oryan A, Davarmanesh M, Shirian S, Negahban S, Aledavood A, et al. Clinical, histopathologic, and cytologic diagnosis of mucosal leishmaniosis and literature review. Arch Pathol Lab Med 2011; 135(4): 478-482.

[4] Nylén S, Eidsmo L. Tissue damage and immunity in cutaneous leishmaniosis. Parasite Immunol 2012; 34(12): 551-561.

[5] Rahmanian V, Bokaie S, Haghdoost A, Barouni M. Predicting cutaneous leishmaniasis using SARIMA and Markov switching models in Isfahan, Iran: A time-series study. Asian Pac J Trop Med 2021; 14(2): 83-93.

[6] Fonseca-Silva F, Inacio JD, Canto-Cavalheiro MM, Almeida-Amaral EE. Reactive oxygen species production and mitochondrial dysfunction contribute to quercetin induced death in Leishmania amazonensis. PLoS One 2011; 6(2): e14666.

[7] Tasdemir D, Kaiser M, Brun R, Yardley V, Schmidt TJ, Tosun F, et al. Antitrypanosomal and antileishmanial activities of flavonoids and their analogues: In vitro, in vivo, structure-activity relationship, and quantitative structure-activity relationship studies. Antimicrob Agents Chemother 2006; 50(4): 1352-1364.

[8] Manjolin LC, dos Reis MBG, do Carmo Maquiaveli C, Santos-Filho OA, da Silva ER. Dietary flavonoids fisetin, luteolin and their derived compounds inhibit arginase, a central enzyme in Leishmania (Leishmania) amazonensis infection. Food Chem 2013; 141(3): 2253-2262.

[9] Chondrogianni N, Kapeta S, Chinou I, Vassilatou K, Papassideri I, Gonos ES. Anti-aging and rejuvenating effects of quercetin. Exp Gerontol 2010; 45(10): 763-771.

[10] Ertuğ PU, Aydinoglu F, Ozturk OG, Singirik E, Ögülener N. Comparative study of the quercetin, ascorbic acid, glutathione and superoxide dismutase for nitric oxide protecting effects in mouse gastric fundus. Eur J Pharmacol 2013; 698(1-3): 379-387.

[11] Lee GH, Lee SJ, Jeong SW, Kim HC, Park GY, Lee SG, et al. Anti-oxidative and antiinflammatory activities of quercetin-loaded silica nanoparticles. Colloids Surf B Biointerfaces 2016; 143: 511-517.

[12] Lin YS, Tsai PH, Kandaswami CC, Cheng CH, Ke FC, Lee PP, et al. Effects of dietary flavonoids, luteolin, and quercetin on the reversal of epithelial-mesenchymal transition in A431 epidermal cancer cells. Cancer Sci 2011; 102(10): 1829-1839.

[13] Sen G, Biswas D, Ray M, Biswas T. Albumin-quercetin combination offers a therapeutic advantage in the prevention of reduced survival of erythrocytes in visceral leishmaniosis. Blood Cells Mol Dis 2007; 39(3): 245-254.

[14] Belkhelfa-Slimani R, Djerdjouri B. Caffeic acid and quercetin exert caspases-independent apoptotic effects on Leishmania major promastigotes, and reactivate the death of infected phagocytes derived from BALB/c mice. Asian Pac J Trop Biomed 2017; 7(4): 321-331.

[15] da Silva ER, do Carmo Maquiaveli C, Magalhães PP. The leishmanicidal flavonols quercetin and quercitrin target Leishmania amazonensis arginase. Exp Parasitol 2012; 130(3): 183-188.

[16] Bemani E, Oryan A, Bahrami S. Effectiveness of amiodarone in treatment of cutaneous leishmaniasis caused by Leishmania major. Exp Parasitol 2019; 205: 107747.

[17] Lyu SY, Park WB. Production of cytokine and NO by RAW 264.7 macrophages and PBMC in vitro incubation with flavonoids. Arch Pharm Res 2005; 28(5): 573.

[18] Benaim G, Casanova P, Hernandez-Rodriguez V, Mujica-Gonzalez S, Parra-Gimenez N, Plaza-Rojas L, et al. Dronedarone, an amiodarone analog with improved anti-Leishmania mexicana efficacy. Antimicrob Agents Chemother 2014; 58(4): 2295-2303.

[19] Bahrami S, Esmaeilzadeh S, Zarei M, Ahmadi F. Potential application of nanochitosan film as a therapeutic agent against cutaneous leishmaniosis caused by L. major. Parasitol Res 2015; 114(12): 4617-4624.

[20] Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Anal Biochem 1996; 239(1): 70-76.

[21] Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem 1966; 16(2): 359-364.

[22] Abdoli A, Maspi N, Ghaffarifar F. Wound healing in cutaneous leishmaniosis: A double edged sword of IL-10 and TGF-β. Comp Immunol Microbiol Infect Dis 2017; 51: 15-26.

[23] Oryan A, Shirian S, Tabandeh MR, Hatam GR, Randau G, Daneshbod Y. Genetic diversity of Leishmania major strains isolated from different clinical forms of cutaneous leishmaniosis in southern Iran based on minicircle kDNA. Infect Genet Evol 2013; 19: 226-231.

[24] Scorza BM, Carvalho EM, Wilson ME. Cutaneous manifestations of human and murine leishmaniosis. Int J Mol Sci 2017; 18(6): 1296.

[25] Oryan A. Plant-derived compounds in treatment of leishmaniosis. Iran J Vet Res 2015; 16(1): 1-19.

[26] Sundar S, Olliaro PL. Miltefosine in the treatment of leishmaniosis: Clinical evidence for informed clinical risk management. Ther Clin Risk Manag 2007; 3: 733-740.

[27] Sousa-Batista AJ, Poletto FS, Philipon CIMS, Guterres SS, Pohlmann AR, Rossi-Bergmann B. Lipid-core nanocapsules increase the oral efficacy of quercetin in cutaneous leishmaniosis. Parasitology 2017; 144(13): 1769-1774.

[28] Sakthianandeswaren A, Elso CM, Simpson K, Curtis JM, Kumar B, Speed TP, et al. The wound repair response controls outcome to cutaneous leishmaniosis. Proc Natl Acad Sci 2005; 102(43): 15551-15556.

[29] Ashcroft GS, Jeong MJ, Ashworth JJ, Hardman M, Jin W, Moutsopoulos N, et al. Tumor necrosis factor-alpha (TNF-α) is a therapeutic target for impaired cutaneous wound healing. Wound Repair Regen 2012; 20(1): 38-49.

[30] Ivonise F, Cibele A, Olívia B, Clarissa BA, Lucas PC, Roque PA, et al. Epidemiologic and immunologic findings for the subclinical form of Leishmania braziliensis infection. Clin Infect Dis 2002; 34(11): 54-58.

[31] Rostamian M, Niknam HM. Role of higher levels of post-challenge antibodies in protective vaccination against Leishmania tropica infection of BALB/c mice. Asian Pac J Trop Biomed 2020; 10(12): 532-539.

[32] Araujo CF, Oliveira IB, Silveira MB, Ribeiro-Dias F. Disseminated cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis in human immunodeficiency virus (HIV)-infected patients: A report of two cases. Asian Pac J Trop Med 2021; 14(6): 281-284.

[33] Padigel UM, Alexander J, Farrel JP. The role of interleukin-10 in susceptibility of BALB/c mice to infection with Leishmania mexicana and Leishmania amazonensis. J Immunol 2003; 171(7): 3705-3710.

[34] Liu J, Li X, Yue Y, Li J, He T, He Y. The inhibitory effect of quercetin on IL-6 production by LPS-stimulated neutrophils. Cell Mol Immunol 2005; 2(6): 455-460.

[35] Kaplanski G, Marin V, Montero-Julian F, Mantovani A, Farnarier C. IL-6: A regulator of the transition from neutrophil to monocyte recruitment during inflammation. Trends Immunol 2003; 24(1): 25-29.

[36] Oliveira F, Bafica A, Rosato AB, Favali CB, Costa JM, Cafe V, et al. Lesion size correlates with Leishmania antigen-stimulated TNF-levels in human cutaneous leishmaniosis. Am J Trop Med Hyg 2011; 85(1): 70-73.

[37] Liew F, Li Y, Millott S. Tumor necrosis factor-alpha synergizes with IFN-gamma in mediating killing of Leishmania major through the induction of nitric oxide. J Immunol 1990; 145(12): 4306-4310.

[38] Hakimi S, Rivière S, Del Giudice P, Dereure J, Le Quellec A. Localized cutaneous leishmaniosis due to Leishmania infantum in a patient treated with infliximab. Dermatology 2010; 220(1): 63-65.

[39] Gopalakrishnan A, Ram M, Kumawat S, Tandan S, Kumar D. Quercetin accelerated cutaneous wound healing in rats by increasing levels of VEGF and TGF-β1. Indian J Exp Biol 2016; 54: 187-195.

[40] Bruun JM, Lihn AS, Verdich C, Pedersen SB, Toubro S, Astrup A, et al. Regulation of adiponectin by adipose tissue-derived cytokines: In vivo and in vitro investigations in humans. Am J Physiol Endocrinol Metab 2003; 285(3): 527-533.

[41] Robinson K, Prins J, Venkatesh B. Clinical review: Adiponectin biology and its role in inflammation and critical illness. Crit Care 2011; 15(2): 221.

[42] Shibata S, Tada Y, Asano Y, Hau CS, Kato T, Saeki H, et al. Adiponectin regulates cutaneous wound healing by promoting keratinocyte proliferation and migration via the ERK signaling pathway. J Immunol 2012; 189(6): 3231-3241.

[43] Wijnant GJ, Van Bocxlaer K, Yardley V, Murdan S, Croft SL. Efficacy of paromomycin-chloroquine combination therapy in experimental cutaneous leishmaniasis. Antimicrob Agents Chemother 2017; 61(8). doi: 10.1128/AAC.00358-17.

[44] Carneiro PP, Conceição J, Macedo M, Magalhães V, Carvalho EM, Bacellar O. The role of nitric oxide and reactive oxygen species in the killing of Leishmania braziliensis by monocytes from patients with cutaneous leishmaniosis. PLoS One 2016; 11(2): e0148084.

[45] Jafari M, Shirbazou S, Norozi M. Induction of oxidative stress in skin and lung of infected BALB/C mice with Iranian strain of Leishmania major (MRHO/IR/75/ER). Iran J Parasitol 2014; 9(1): 60.