|Year : 2017 | Volume
| Issue : 1 | Page : 43-48
The evaluation of quercetin and luteolin efficacy on cutaneous leishmaniasis in mice infected with Leishmania major
Nasrin Hamidizadeh1, Sara Ranjbar1, Qasem Asgari2, Gholamreza Hatam3
1 Molecular Dermatology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
2 Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
3 Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz; Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
|Date of Web Publication||21-Apr-2017|
Basic Sciences in Infectious Diseases Research Center, School of medicine, Shiraz University of Medical Sciences, Shiraz
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Introduction: Almost 1.5-2 million people are infected with leishmaniasis and 60,000 die due to the disease every year. The treatment of cutaneous leishmaniasis with the existing medications is not effective and accompanied with various side effects. Expensiveness and not being available in different forms are other limitations associated with these drugs. In addition to, parasite resistance restricts the use of these medications; therefore, identification of new treatment methods is highly essential. Humans have always used medicinal plants for the treatment of diseases. This study aimed to assess the impact of quercetin and luteolin on leishmaniasis wounds created on mice's tails. Materials and Methods: A total of 56 infected Balb/C mice were divided into eight groups, each containing seven animals. The first three groups received 3.5 mg/kg luteolin through oral, intradermal, and intraperitoneal routes. The other three groups received 14 mg/kg quercetin through oral, intradermal, and intraperitoneal routes. The treatments were administered twice a week for 4 weeks in comparison to meglumine antimonite as control groups. Results: The results showed that there was no statistically significant difference between the recovery of mice in the luteolin and quercetin groups and those in the meglumine antimonite group. However, the ratio of recovered Balb/C mice in the luteolin and quercetin group is 1.75-2 times more than meglumine group, respectively. Conclusion: Quercetin and luteolin seems to be candidate medications with fewer side effects for recovery from cutaneous leishmaniasis. They can also be used as complementary medications together with other standard drugs, such as meglumine, antimonite, and cryotherapy.
Keywords: Cutaneous leishmaniasis, luteolin, mice, quercetin
|How to cite this article:|
Hamidizadeh N, Ranjbar S, Asgari Q, Hatam G. The evaluation of quercetin and luteolin efficacy on cutaneous leishmaniasis in mice infected with Leishmania major. J Pharm Negative Results 2017;8:43-8
|How to cite this URL:|
Hamidizadeh N, Ranjbar S, Asgari Q, Hatam G. The evaluation of quercetin and luteolin efficacy on cutaneous leishmaniasis in mice infected with Leishmania major. J Pharm Negative Results [serial online] 2017 [cited 2018 Jan 16];8:43-8. Available from: http://www.pnrjournal.com/text.asp?2017/8/1/43/204905
| Introduction|| |
Cutaneous leishmaniasis is an endemic disease in Iran and across the world, which is common to both humans and animals. In Iran, leishmaniasis is created in mammals' cells by Leishmania parasite that belongs to Kinetoplastida order and Trypanosomatidae family. Up to now, 30 Leishmania species have been identified, out of which 21 are capable of infecting humans. Almost 1.5-2 million individuals are infected with leishmaniasis and 60,000 die due to disease every year. However, this disease has been neglected among tropical diseases (WHO, 2010). To date, nearly 12 million individuals around the world are infected with the parasite and the incidence of the infection is increasing in local areas (WHO, 2009).Leishmania tropica and Leishmania major are two of the most common Leishmania species distributed in more than 18 out of 31 provinces in Iran. Leishmania major is the most frequent species in Fars province, south of Iran. The number of cases infected by leishmaniasis in this region has risen sharply in recent years. During 2001-2008, the cases reported in Fars province comprised approximately 25% of the total number of cases (44,464 cases) with CL in the country. From 2005 to 2008, the number of cases increased from 192 to 484 in Shiraz, the capital of Fars province., A total of three species of Leishmania parasites were identified in this area, including L. major, L. tropica, and dermotropic L. infantum.
In modern medicine, various medications are used to treat cutaneous leishmaniasis. For instance, antimonial (V) compounds, sodium stibogluconate, and N- methylglucamine are the first medications used for the treatment of leishmaniasis, which inhibit specific enzymes in the parasite's metabolism., However, these medications are accompanied with serious side-effects and their effectiveness is questionable. On the contrary, amphotericin and pentamidine comprise the second-line therapy of leishmaniasis, which are not effective permanently.,,,, In addition, they have serious toxic side effects, such as heart and kidney failure. Paromomycin is also used for the treatment of cutaneous leishmaniasis. Miltefosine, which has teratogenic side effects, has also been utilized for the treatment of kala azar  and cutaneous leishmaniasis since 2002. All these medications are expensive and not available in different forms. Additionally, parasite resistance further restricts the use of these drugs. Therefore, identification of new therapeutic methods is highly essential. Despite what has mentioned above, leishmaniasis still remains a general health concern all over the world.
Humans have always benefited from medicinal plants to treat diseases. Indeed, Iran has been home to many great scientists who commonly used medicinal plants. Nevertheless, most beneficial medicinal plants have not been identified in our country. The identified ones are also mainly used in traditional rather than modern medicine., Since 1994, many medicinal plants have been found to be effective in the treatment of leishmaniasis in vivo and in vitro. Some in-vivo and in-vitro studies were also conducted on the effects of medicinal plants on cutaneous leishmaniasis,,,,, as well as on leishmaniasis wounds in mice., The results of these studies indicated reduction in Leishmania growth, number, and leishmaniasis wound diameter.
Quercetin and luteolin are the main members of flavonoids family, which are found in many fruits, vegetables, olive oil, red wine, tea, and beeswax and are active against some Leishmania species.,,,
In addition to various therapeutic uses, quercetin inhibits development of leukemia cells, Ehrlich ascites tumor cells, and other ascites.,,, It also stimulates creation of breaches in specific locations on mammal cells' DNAs through topoisomerase in vitro., In fact, topoisomerase inhibition results in cell death. Evidence has demonstrated that luteolin and quercetin inhibited the catalyzer activity of DNA topoisomerase II in leishmaniasis. Thus, it can be speculated that flavonoids exert their antileishmaniasis effects through topoisomerase inhibition, which is similar to pentamidine and anticancer medications, such as etoposides. Then, topoisomerase inhibition stimulates disruption of cell cycle and results in L. donovani promastigotes apoptosis. In this context, researchers examined the treatment potentiality of these two flavonoids in animal models. The findings revealed that luteolin is similar to quercetin, but in lower concentrations, considerably decreases spleen parasites. Luteolin seems to be nontoxic and, consequently, might be accompanied with better antileishmaniasis effects in case used together with classical treatment methods, acts anti-inflammatory and anticarcinogenic. Therefore, the present study aims to assess the impact of quercetin and luteolin on leishmaniasis wounds on mice's tails.
| Materials and Methods|| |
Luteolin and quercetin powder were obtained from Sigma-Aldrich Company, Germany. In this study, 56 Balb/C mice (4-6-weeks-old, 15 g) were bought from Pasteur Institute, Karaj, Iran. After making sure about the mice's conditions through completion of work with laboratory animals' checklist, the mice were divided into eight groups, each containing seven animals. Then, amastigotes from L. major parasite from infected mice were intradermally injected to the mice's tails. After appearance of wounds, the wound areas were photographed. The first three groups received 3.5 mg/kg luteolin, whereas the second three groups received 14 mg/kg quercetin through oral, intradermal, and intraperitoneal routes twice a week for 4 weeks. The wounds' status was followed up by taking photos every week for 15 weeks. The control group also received 20 mg/kg meglumine antimonite, intraperitoneally, five times a week, and the placebo group received phosphate-buffered saline (PBS), intraperitoneally, twice a week for 4 weeks. The results were also evaluated by Fisher's exact test due to the small number of mice in each group.
| Results|| |
According to [Table 1], Balb/C mice were divided into the therapeutic groups and the status of their recovery and death are as follows.
Although equal number of mice was entered into the study groups, some mice were excluded due to small nodules, which was hard to be confirmed in photos.
As it is displayed in [Table 2], there is no statistically significant difference between the recovery of mice in luteolin and quercetin groups and those in meglumine antimonate (cutaneous leishmaniasis standard drug) and placebo groups (p-value = 0.353).
|Table 2: The comparison of recovery and mortality of Balb/C mice between four therapeutic groups including luteolin, quercetin, meglumine antimonate, and placebo groups|
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In [Table 3], luteolin is compared with meglumine. There is no statistically significant difference in cutaneous leishmaniasis recovery between the Balb/C mice in luteolin group and those in meglumine antimonate group (P value = 0.606).
|Table 3: The comparison of recovery and mortality of Balb/C mice between luteolin and meglumine antimonate groups|
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[Table 4] also shows no significant difference between quercetin and meglumine antimonate in the recovery of Balb/C mice from cutaneous leishmaniasis (P value = 1.00).
|Table 4: The comparison of recovery and mortality of Balb/C mice between querctin and meglumine antimonate groups|
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| Disussion|| |
Every year, 1.5-2 million individuals are infected with leishmaniasis. Cutaneous leishmaniasis can be treated by antimonial (V) compounds,, amphotericin, pentamidine,,, paramomycin, and miltefosine, which have toxic side effects, such as heart and kidney failure and teratogenic complications. In addition, they are expensive, are not available in different forms, and are not effective permanently.,, Overall, 60,000 patients with cutaneous leishmaniasis die annually. Therefore, identification of effective medications with lower side effects is of utmost importance. In this context, medicinal plants are accompanied with lower side effects. The impact of medicinal plants on leishmaniasis in mice has been confirmed in both in-vivo and in-vitro studies. In this study, we assess the effects of quercetin and luteolin on leishmaniasis wounds on mice's tails.
Quercetin and luteolin belong to flavonoid family and exist in many fruits, vegetables, olive oil, red wine, tea, and beeswax.
Quercetin inhibits development of leukemia cells, Ehrlich ascites tumor cells, and other ascites. It also stimulates creation of breaches in specific locations on mammal cells' DNAs through topoisomerase in vitro. Topoisomerase inhibition, in turn, results in cell death.,, In-vitro investigations have indicated that luteolin and quercetin exert their antileishmaniasis effects through inhibition of the catalyzer activity of DNA topoisomerase II, which stimulates disruption of cell cycle and results in L. donovani promastigotes apoptosis. Further, luteolin similar to quercetin, but in lower concentrations, considerably decreased spleen parasites and had better antileishmaniasis effects in case used together with classical treatment methods.
Luteolin and quercetin are L. donovani topoisomerase I inhibitors, topoisomerase inhibition leads to apoptosis. Ethanol extract of Guazima ulmifolia is a powerful antileishmanial compound due to its quercetin. An in-vitro study by Tasdemir et al. showed that quercetin and its derivatives have strong antiparasitic effect on L. donovani. The substantial inhibitory effect of luteolin and quercetin on the growth of promastigotes and amastigotes was shown in another in vitro study. Quercetin and luteolin can have antileishmanial effect on L. amazonensis by inhibiting arginase enzyme and can be used as oral complementary medicines for treatment of Leishmania.
The recent study on the effect of luteolin and quercetin on Balb/C mice's recovery from cutaneous leishmaniasis showed that there is no statistically significant difference between mice's recovery in the luteolin and quercetin groups and that of those in the meglumine antimonate group [Table 2]. But the ratio of recovered Balb/C mice in the quercetin and luteolin groups is 1.75-2 times more than those in meglumine antimonate group [Table 3], [Table 4], and [Table 5]. This result can be due to low sample size. It is likely that if the sample size enlarges, significant differences will be observed between different groups in response to treatment. Moreover, no significant difference was observed between the treatment methods (data not shown).
|Table 5: The comparison of recovery and mortality of Balb/C mice between luteolin and quercetin groups and those in the meglumine antimonate group|
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However, the comparison of the drug intraperitoneal administration methods indicates that the percentage of mortality in those mice which received luteolin and quercetin through intraperitoneal routes was lower as compared with those in control group (intraperitoneal meglumine antimonate). This research showed that luteolin and quercetin can be effective in cutaneous leishmaniasis recovery. The use of these two drugs as effective medications for cutaneous leishmaniasis treatment requires more studies with larger sample sizes. Additionally, these medications accompanied with classical treatment methods, such as meglumine antimonate, and cryotherapy can be more effective.
| Conclusion|| |
Cutaneous leishmaniasis is a complex disease and it is not possible to find a quite effective medication for it easily. According to previous studies on the antileishmanisis effects of luteolin and quercetin as well as the effect of these two drugs on cutaneous leishmaniasis recovery found in this study, these two medicines can be used as candidate medicines with fewer or without side effects for patients' recovery from cutaneous leishmaniasis, provided that more extensive studies with larger sample sizes will be conducted or they may be used as complementary medicines, accompanied with other standard treatment methods. Thus, simultaneous administration of these two medications might be accompanied with better results. They can also be used together with other standard treatment, such as meglumine antimonate and cryotherapy.
The authors would like to thank Mrs. Shahrbanou Naderi, The Vice Chancellors of Research of Shiraz University of Medical Sciences for financial support, Shiraz, Iran, Department of Parasitology and Mycology, Shiraz University of Medical Sciences and Omid Kouhi, Center of Comparative and Experimental Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Lainson R, Shaw JJ, Silveira FT. Dermal and visceral leishmaniasis and their causative agents. Trans R Soc Trop Med Hyg 1987;81:702-3.
Ashford RW. The leishmaniases as emerging and reemerging zoonoses. Int J Parasitol 2000;30:1269-81.
Shirzadi MR. Neglected tropical diseases innovative and intensified disease management leishmaniasis control. WHO Headquarters report of the consultative meeting on cutaneous leishmaniasis. April, 2007
Razmjou S, Hejazy H, Motazedian MH, Baghaei M, Emamy M, Kalantary M. A new focus of zoonotic cutaneous leishmaniasis in Shiraz, Iran. Trans R Soc Trop Med Hyg 2009;103:727-30.
Oshaghi MA, Rasoolian M, Shirzadi MR, Doosti LS. First report on isolation of Leishmania tropica from sandflies of a classical urban cutaneous leishmaniasis focus in southern Iran. Exp Parasitol 2010;126:445-50.
Pourmohammadi B, Motazedian M, Hatam G, Kalantari M, Habibi P, Sarkar B. Comparison of three methods for diagnosis of cutaneous leishmaniasis. Iran J Parasitol 2010;5:1-8.
Kobets T, Grekov I, Lipoldova M. Leishmaniasis: prevention, parasite detection and treatment. Curr Med Chem 2012;19:1443-74.
Berman JD, Gallalee JV, Best JM. Sodium stibogluconate (Pentostam) inhibition of glucose catabolism via the glycolytic pathway, and fatty acid beta-oxidation in Leishmania
mexicana amastigotes. Biochem Pharmacol 1987;36:197-201.
Wyllie S, Cunningham ML, Fairlamb AH. Dual action of antimonial drugs on thiol redox metabolism in the human pathogen Leishmania
donovani. J Biol Chem 2004;279:39925-32.
Pourmohammadi B, Motazedian MH, Handjani F, Hatam GHR, Habibi S, Sarkari B. Glucantime efficacy in the treatment of zoonotic cutaneous leishmaniasis. Southeast Asian J Trop Med Public Health 2011;42:502-8.
Croft SL. Recent developments in the chemotherapy of leishmaniasis. Trends Pharmacol Sci 1988;9:376-81.
Olliaro PL, Bryceson AD. Practical progress and new drugs for changing patterns of leishmaniasis. Parasitol Today 1993;9:323-8.
Bryceson ADM. Therapy in man. The Leishmaniase in Biology and Medicine. London: Academic Press; 1987. p. 848-907
Mebrahtu Y, Lawyer P, Githure J, Were JB, Muigai R, Hendricks L, et al. Visceral leishmaniasis unresponsive to pentostam caused by Leishmania
tropica in Kenya. Am J Trop Med Hyg 1989 Sep;41:289-94.
Anon TDR News. World Health Organization 1990;
el-Safi SH, Murphy AG, Bryceson AD, Neal RA. A double-blind clinical trial of the treatment of cutaneous leishmaniasis with paromomycin ointment. Trans R Soc Trop Med Hyg 1990;84:690-1.
Sundar S, Jha TK, Thakur CP, Engel J, Sindermann H, Fischer C, et al. Oral miltefosine for Indian visceral leishmaniasis. N Engl J Med 2002;347:1739-46.
Soto J, Toledo J, Gutierrez P, Nicholls RS, Padilla J, Engel J, et al. Treatment of American cutaneous leishmaniasis with miltefosine, an oral agent. Clin Infect Dis 2001;33:E57-61.
Shariatifar N, Chamanzari H, Ghanay M. The study of flos plant on progmastigote in culture. Horizon Med Sci 2006;11:5-9.
Mohseni N, Sajjadi SE, Eskandarian AA, Yousefi HA, Mansurian M, Shokoohinia Y, et al. Natural anti-leishmaniasis compounds in traditional Iranian medicine. J Islam Iran Trad Med 2012;3:41-50.
Iwu MM, Jackson JE, Schuster BG. Medicinal plants in the fight against leishmaniasis. Parasitol Today 1994;10:65-8.
Azizi K, Shahidi-Hakak F, Asgari Q, Hatam GR, Fakoorziba MR, Miri R, et al. In vitro
efficacy of ethanolic extract of Artemisia absinthium (Asteraceae) against Leishmania
major L. using cell sensitivity and flow cytometry assays. J Parasit Dis 2016;40:735-40.
Moein M, Hatam G, Taghavi-Moghadam R, Zarshenas MM. Antileishmanial Activities of Greek Juniper (Juniperus excelsa M.Bieb.) Against Leishmania
major Promastigotes. J Evid Based Complementary Altern Med 2017;22:31-6.
Barati M, Sharifi I, Sharififar F. Antileishmanial activity of Artemisia aucheri, Ferula asa-foetid and Gossypium hirsutum extracts on Leishmania
major promastigotes in vitro
. HBI-J Army Univ 2010;8:166-72.
Fata A, Rakhshandeh H, Berenji F, Jalalianfard A. Treatment of cutaneous leishmaniasis in murine model by alcoholic extract of berberis vulgaris. Iran J Parasitol 2006;1:39-42.
Kazemi E, Talari S, Hooshyar H. The effect of an alcoholic extract of Berberis Vulgaris on Cutaneous leishmaniasis (L. major) in Balb/C mice. Sci J School Public Health Institute Public Health Res 2007;5:35-42.
Herrmann K. Flavonols and flavonones in food plants: a review. J Food Technol 1976;11:433-48.
Sen G, Mukhopadhyay S, Ray M, Biswas T. Quercetin interferes with iron metabolism in Leishmania
donovani and targets ribonucleotide reductase to exert leishmanicidal activity. J Antimicrob Chemother 2008;61:1066-75.
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;6:527-41.
Muzitano MF, Tinoco LW, Guette C, Kaiser CR, Rossi-Bergmann B, Costa SS. The antileishmanial activity assessment of unusual flavonoids from Kalanchoe pinnata. Phytochemistry 2006;67:2071-7.
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;50:1352-64.
Suolinna EM, Buchsbaum RN, Racker E. The effect of flavonoids on aerobic glycolysis and growth of tumor cells. Cancer Res 1975;35:1865-72.
Castillo MH, Perkins E, Campbell JH, Doerr R, Hassett JM, Kandaswami C, et al. The effects of the bioflavonoid quercetin on squamous cell carcinoma of head and neck origin. Am J Surg 1989;158:351-5.
Bibby MC, Double JA. Flavone acetic acid-from laboratory to clinic and back. Anticancer Drugs 1993;4:3-17.
Yamashita Y, Kawada S, Fujii N, Nakano H. Induction of mammalian DNA topoisomerase II dependent DNA cleavage by antitumor antibiotic streptonigrin. Cancer Res 1990;50:5841-4.
Austin CA, Patel S, Ono K, Nakane H, Fisher LM. Site-specific DNA cleavage by mammalian DNA topoisomerase II induced by novel flavone and catechin derivatives. Biochem J 1992;282:883-9.
Del Bino G, Bruno S, Yi PN, Darzynkiewicz Z. Apoptotic cell death triggered by camptothecin or teniposide. The cell cycle specificity and effects of ionizing radiation. Cell Prolif 1992;25:537-48.
Seelinger G, Merfort I, Wölfle U, Schempp CM. Anti-carcinogenic effects of the flavonoid luteolin. Molecules 2008;13:2628-51.
Das BB, Sen N, Roy A, Dasgupta SB, Ganguly A, Mohanta BC, et al. Differential induction of Leishmania
donovani bi-subunit topoisomerase I-DNA cleavage complex by selected flavones and camptothecin: activity of flavones against camptothecin-resistant topoisomerase I. Nucleic Acids Res 2006;34:1121-32.
Manjolin LC, dos Reis MB, Maquiaveli Cdo 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;141:2253-62.
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]