Abstract
Honey bee glue (HBG) or propolis is produced by honey bees and used as cementing material for sealing within the hive which is considered as a waste by the beekeepers. Due to its umpteen biological properties, it is having the potential for drug development against infectious diseases. The management of leishmaniasis relied wholly on injectable chemotherapeutic agents with a lengthy regimen and no vaccine is available. Moreover, the growing drug resistance alarming the entire community about the upcoming menace. Hence, it is of utmost importance to discover and explore new compounds against Leishmania infection, which are effective, safe, and cheaper. The present study was aimed to evaluate the efficacy of Indian honey bee glue (HBG), a natural, resinous by-product of the apiculture industry against the promastigote and amastigote forms of the sensitive and resistant strains of Leishmania donovani. HBG was characterized through GC-MS screening and phytochemical (qualitative and quantitative) analysis was done. In vitro (anti-promastigote) and ex vivo (anti-amastigote) activities against sensitive and resistant strains of L. donovani were assessed. Cytotoxicity, ROS (reactive oxygen species) generation, cell membrane integrity, and cell cycle analysis were evaluated. Chemical characterization by GC-MS revealed the presence of different biologically active components in honey bee glue (HBG). Our results showed the activity of HBG with an IC50 of 27.95 ± 1.53 μg/mL and 29.43 ± 1.54 μg/mL against the sensitive and resistant L. donovani promastigotes respectively. An IC50 of 20.93 ± 1.35 μg/mL and 20.22 ± 1.53 μg/mL was observed against the sensitive and resistant amastigotes of L. donovani. Moreover, the cytotoxicity test against the human cell line showed a CC50 value of 811.30 ± 73.44 μg/mL. In addition to this, it enhanced the ROS production and oxidative stress exhibited by the altered levels of lipid peroxidation, superoxide radicals, lipid bodies, and cell membrane integrity within the parasitic cells and mediated their killing by arresting the DNA in the sub-G0/G1 phase of the cell cycle. The antiparasitic action together with GC-MS data revealed the importance of this by-product from the apiary industry. Therefore, HBG can be a good option to develop a new drug molecule against the deadly infection of Leishmania eventually.
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Abbreviations
- HBG:
-
Honey bee glue
- SSG:
-
Sodium stibogluconate
- L. donovani :
-
Leishmania donovani
- RPMI:
-
Roswell Park Memorial Institute
- DMSO:
-
Dimethyl sulfoxide
- PBS:
-
Phosphate-buffered saline
- GC-MS:
-
Gas chromatography-mass spectrometry
- ROS:
-
Reactive oxygen species
- VL:
-
Visceral leishmaniasis
- IC50 :
-
Inhibitory concentration (50%)
- CC50 :
-
Cytotoxic concentration (50%)
- SI:
-
Selectivity index
- B.O.D:
-
Biological oxygen demand
- h:
-
Hour
- min:
-
Minutes
- mL:
-
Millilitres
- μL:
-
Micro litres
- Abs:
-
Absorbance
References
Abdel Moneim AE (2016) Indigofera oblongifolia prevents lead acetate-induced hepatotoxicity, oxidative stress, fibrosis and apoptosis in rats. PLoS One 11:1–18. https://doi.org/10.1371/journal.pone.0158965
Airen B, Sarkar PA, Tomar U, Bishen KA (2018) Antibacterial effect of propolis derived from tribal region on Streptococcus mutans and lactobacillus acidophilus: an in vitro study. J Indian Soc Pedod Prev Dent. https://doi.org/10.4103/JISPPD.JISPPD_1128_17
Alemayehu B, Alemayehu M (2017) Leishmaniasis: a review on parasite, vector and reservoir host. Health Sci J 11:1–6. https://doi.org/10.21767/1791-809x.1000519
Alotaibi A, Ebiloma GU, Williams R, Alenezi S, Donachie AM, Guillaume S, Igoli JO, Fearnley J, de Koning HP, Watson DG (2019) European propolis is highly active against trypanosomatids including Crithidia fasciculata. Sci Rep 9:1–10. https://doi.org/10.1038/s41598-019-47840-y
Antwi CA, Amisigo CM, Adjimani JP, Gwira TM (2019) In vitro activity and mode of action of phenolic compounds on Leishmania donovani. PLoS Negl Trop Dis 13:1–22. https://doi.org/10.1371/journal.pntd.0007206
Asfaram S, Fakhar M, Keighobadi M, Akhtari J (2020) Promising anti-protozoan activities of Propolis (bee glue) as natural product: A review. Acta Parasitol. https://doi.org/10.1007/s11686-020-00254-7
Bankova V, Bertelli D, Borba R, Conti BJ, da Silva Cunha IB, Danert C, Eberlin MN, Falcão IS, Isla MI, Moreno MIN, Papotti G, Popova M, Santiago KB, Salas A, Sawaya ACHF, Schwab NV, Sforcin JM, Simone-Finstrom M, Spivak M et al (2019) Standard methods for Apis mellifera propolis research. J Apic Res 58:1–49. https://doi.org/10.1080/00218839.2016.1222661
Brown HL, Roberts AEL, Cooper R, Jenkins RE (2016) A review of selected bee products as potential anti-bacterial, anti-fungal, and anti-viral agents. Med Res Arch 4:1–20 https://esmed.org/MRA/mra/article/view/887
Bueno-Silva B, Marsola A, Ikegaki M, Alencar SM, Rosalen PL (2017) The effect of seasons on Brazilian red propolis and its botanical source: chemical composition and antibacterial activity. Nat Prod Res 31:1318–1324. https://doi.org/10.1080/14786419.2016.1239088
Cataneo AHD, Tomiotto-Pellissier F, Miranda-Sapla MM, Assolini JP, Panis C, Kian D, Yamauchi LM, Colado Simão AN, Casagrande R, Pinge-Filho P, Costa IN, Verri WA, Conchon-Costa I, Pavanelli WR (2019) Quercetin promotes antipromastigote effect by increasing the ROS production and anti-amastigote by upregulating Nrf2/HO-1 expression, affecting iron availability. Biomed Pharmacother 113:108745. https://doi.org/10.1016/j.biopha.2019.108745
Choudhari MK, Haghniaz R, Rajwade JM, Paknikar KM (2013) Anticancer activity of Indian stingless bee propolis: an in vitro study. Evid Based Complement Alternat Med. https://doi.org/10.1155/2013/928280
Chouhan G, Islamuddin M, Want MY, Abdin MZ, Ozbak HA, Hemeg HA, Sahal D, Afrin F (2015) Apoptosis mediated leishmanicidal activity of Azadirachta indica bioactive fractions is accompanied by Th1 immunostimulatory potential and therapeutic cure in vivo. Parasit Vectors 8:1–24. https://doi.org/10.1186/s13071-015-0788-3
Cortes S, Bruno de Sousa C, Morais T, Lago J, Campino L (2020) Potential of the natural products against leishmaniasis in Old World - a review of in-vitro studies. Pathog Glob Health 114:170–182. https://doi.org/10.1080/20477724.2020.1754655
Crane E (1999) The world history of beekeeping and honey hunting, 1st edn. Taylor & Francis Group, New York
Croft SL, Olliaro P (2011) Leishmaniasis chemotherapy-challenges and opportunities. Clin Microbiol Infect 17:1478–1483. https://doi.org/10.1111/j.1469-0691.2011.03630.x
da Silva Rodrigues JH, Miranda N, Volpato H, Ueda-Nakamura T, Nakamura CV (2019) The antidepressant clomipramine induces programmed cell death in Leishmania amazonensis through a mitochondrial pathway. Parasitol Res 118:977–989. https://doi.org/10.1007/s00436-018-06200-x
Dantas AP, Barbosa HS, De Castro SL (2003) Biological and ultrastructural effects of the anti-microtubule agent taxol against Trypanosoma cruzi. J Submicrosc Cytol Pathol 35:287–294 PMID: 14690177 Cite
Dantas AP, Salomão K, Barbosa HS, De Castro SL (2006) The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells. Mem Inst Oswaldo Cruz 101:207–211. https://doi.org/10.1590/S0074-02762006000200013
Das R, Roy A, Dutta N, Majumder HK (2008) Reactive oxygen species and imbalance of calcium homeostasis contributes to curcumin induced programmed cell death in Leishmania donovani. Apoptosis 13:867–882. https://doi.org/10.1007/s10495-008-0224-7
Das A, Das MC, Das N, Bhattacharjee S (2017a) Evaluation of the antileishmanial potency, toxicity and phytochemical constituents of methanol bark extract of Sterculia villosa. Pharm Biol 55:998–1009. https://doi.org/10.1080/13880209.2017.1285946
Das A, Jawed JJ, Das MC, Sandhu P, De UC, Dinda B, Akhter Y, Bhattacharjee S (2017b) Antileishmanial and immunomodulatory activities of lupeol, a triterpene compound isolated from Sterculia villosa. Int J Antimicrob Agents 50:512–522. https://doi.org/10.1016/j.ijantimicag.2017.04.022
De MacEdo Silva ST, Visbal G, Godinho JLP, Urbina JA, De Souza W, Rodrigues JCF (2018) In vitro antileishmanial activity of ravuconazole, a triazole antifungal drug, as a potential treatment for leishmaniasis. J Antimicrob Chemother 73:2360–2373. https://doi.org/10.1093/jac/dky229
Demir S, Aliyazicioglu Y, Turan I, Misir S, Mentese A, Yaman SO, Akbulut K, Kilinc K, Deger O (2016) Antiproliferative and proapoptotic activity of Turkish propolis on human lung cancer cell line. Nutr Cancer 68:165–172. https://doi.org/10.1080/01635581.2016.1115096
Dikhit MR, Kumar A, Das S, Dehury B, Rout AK, Jamal F, Sahoo GC, Topno RK, Pandey K, Das VNR, Bimal S, Das P (2017) Identification of potential MHC class-II-restricted epitopes derived from Leishmania donovani antigens by reverse vaccinology and evaluation of their CD4+ T-cell responsiveness against visceral leishmaniasis. Front Immunol. https://doi.org/10.3389/fimmu.2017.01763
do Nascimento TG, da Silva PF, Azevedo LF, da Rocha LG, de Moraes Porto ICC, e Moura TFAL, Basílio-Júnior ID, Grillo LAM, Dornelas CB, da Silva Fonseca EJ, de Jesus Oliveira E, Zhang AT, Watson DG (2016) Polymeric nanoparticles of Brazilian red Propolis extract: preparation, characterization, Antioxidant and Leishmanicidal Activity. Nanoscale Res Lett. https://doi.org/10.1186/s11671-016-1517-3
El-Khadragy M, Alolayan EM, Metwally DM, El-Din MFS, Alobud SS, Alsultan NI, Alsaif SS, Awad MA, Moneim AEA (2018) Clinical efficacy associated with enhanced antioxidant enzyme activities of silver nanoparticles biosynthesized using Moringa oleifera leaf extract, against cutaneous leishmaniasis in a murine model of Leishmania major. Int J Environ Res Public Health 15:1037. https://doi.org/10.3390/ijerph15051037
Ezeonu CS, Ejikeme CM (2016) Qualitative and quantitative determination of phytochemical contents of indigenous Nigerian softwoods. New J Sci 2016:1–9. https://doi.org/10.1155/2016/5601327
Ghorai N, Ghorai N, Chakraborty S, Gucchait S, Saha SK, Biswas S (2012) Estimation of total Terpenoids concentration in plant tissues using a monoterpene. Linal stand reag Protoc Exch:1–7. https://doi.org/10.1038/protex.2012.055
Goyal DK, Keshav P, Kaur S (2021a) Immune induction by adjuvanted Leishmania donovani vaccines against the visceral leishmaniasis in BALB/c mice. Immunobiology 226:152057. https://doi.org/10.1016/j.imbio.2021.152057
Goyal DK, Keshav P, Kaur S (2021b) Adjuvanted vaccines driven protection against visceral infection in BALB/c mice by Leishmania donovani. Microb Pathog 151:104733. https://doi.org/10.1016/j.micpath.2021.104733
Goyal DK, Keshav P, Kaur S (2021c) Adjuvant effects of TLR agonist gardiquimod admixed with Leishmania vaccine in mice model of visceral leishmaniasis. Infect Genet Evol 93:104947. https://doi.org/10.1016/j.meegid.2021.104947
Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P, Wasunna MK, Bryceson ADM (2002) Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2:494–501. https://doi.org/10.1016/S1473-3099(02)00347-X
Hasanah N, Astuty H (2019) Influence of the combination of Pasak bumi root and propolis extracts on parasitemia levels in mice infected with plasmodium berghei. Int J Appl pharm 11:268–271. https://doi.org/10.22159/ijap.2019.v11s1.015
Kasote DM (2017) Propolis: a neglected product of value in the Indian beekeeping sector. Bee World 94:80–83. https://doi.org/10.1080/0005772x.2017.1345223
Kathuria M, Bhattacharjee A, Sashidhara KV, Singh SP, Mitra K (2014) Induction of mitochondrial dysfunction and oxidative stress in Leishmania donovani by orally active clerodane diterpene. Antimicrob Agents Chemother 58:5916–5928. https://doi.org/10.1128/AAC.02459-14
Kaye P, Scott P (2011) Leishmaniasis : complexity at the host – pathogen interface. Nat Publ Gr 9:604–615. https://doi.org/10.1038/nrmicro2608
Kaye PM, Cruz I, Picado A, Van Bocxlaer K, Croft SL (2020) Leishmaniasis immunopathology—impact on design and use of vaccines, diagnostics and drugs. Semin Immunopathol 42:247–264. https://doi.org/10.1007/s00281-020-00788-y
Keshav P, Goyal DK, Kaur S (2021a) Antileishmanial potential of immunomodulator gallic acid against experimental murine visceral leishmaniasis. Paarasite Immunol. https://doi.org/10.1111/pim.12875
Keshav P, Goyal DK, Kaur S (2021b) GC – MS screening and antiparasitic action of Putranjiva roxburghii leaves against sensitive and resistant strains of Leishmania donovani. J Parasit Dis:1–12. https://doi.org/10.1007/s12639-021-01388-9
Keshav P, Goyal DK, Kaur S (2021c) Promastigotes of Leishmania donovani exhibited sensitivity towards the high altitudinal plant Cicer microphyllum. Curr Res Parasitol Vector Borne Dis 1:100040. https://doi.org/10.1016/j.crpvbd.2021.100040
Kumar S, Pandey AK (2013) Chemistry and biological activities of flavonoids: an overview. Sci World J:1–16 https://doi.org/10.1155/2013/162750
Lee SJ, Zhang J, Choi AMK, Kim HP (2013) Mitochondrial dysfunction induces formation of lipid droplets as a generalized response to stress. Oxidative Med Cell Longev 2013:1–10. https://doi.org/10.1155/2013/327167
Mehta A, Shaha C (2006) Mechanism of metalloid-induced death in Leishmania spp.: role of iron, reactive oxygen species, Ca2+, and glutathione. Free Radic Biol Med 40:1857–1868. https://doi.org/10.1016/j.freeradbiomed.2006.01.024
Misra P, Kumar A, Khare P, Gupta S, Kumar N, Dube A (2009) Pro-apoptotic effect of the landrace Bangla Mahoba of Piper betle on Leishmania donovani may be due to the high content of eugenol. J Med Microbiol 58:1058–1066. https://doi.org/10.1099/jmm.0.009290-0
Morales-Yuste M, Morillas-Márquez F, Martín-Sánchez J, Valero-López A, Navarro-Moll MC (2010) Activity of (−)α-bisabolol against Leishmania infantum promastigotes. Phytomedicine 17:279–281. https://doi.org/10.1016/j.phymed.2009.05.019
Mukherjee D, Singh CB, Dey S, Mandal S, Ghosh J, Mallick S, Hussain A, Swapana N, Ross SA, Pal C (2016) Induction of apoptosis by zerumbone isolated from Zingiber zerumbet (L.) smith in protozoan parasite Leishmania donovani due to oxidative stress. Braz J Infect Dis 20:48–55. https://doi.org/10.1016/j.bjid.2015.10.002
Mutoro CN, Kinyua J, Ng’Ang’A J, Kariuki D, Ingonga JM, Anjili CO (2018) Efficacy of the combination of crude extracts of Solanum nigrum and on plumbago capensis Leishmania major [version 1; peer review: 1 approved, 1 approved with reservations]. F1000Research. https://doi.org/10.12688/F1000RESEARCH.15955.1
Naik DG, Mujumdar AM, Vaidya HS (2013) Anti-inflammatory activity of propolis from Maharashtra, India. J Apic Res 52:35–43. https://doi.org/10.3896/IBRA.1.52.2.06
Oliveira ACP, Shinobu CS, Longhini R, Franco SL, Svidzinski TIE (2006) Antifungal activity of propolis extract against yeasts isolated from onychomycosis lesions. Mem Inst Oswaldo Cruz. https://doi.org/10.1590/S0074-02762006000500002
Onal G, Kutlu O, Gozuacik D, Emre SD (2017) Lipid droplets in health and disease. Lipids Health Dis 16:1–15. https://doi.org/10.1186/s12944-017-0521-7
Panche AN, Diwan AD, Chandra SR (2016) Flavonoids: An overview. J Nutr Sci. https://doi.org/10.1017/jns.2016.41
Park YS, Jung ST, Kang SG, Heo BG, Arancibia-Avila P, Toledo F, Drzewiecki J, Namiesnik J, Gorinstein S (2008) Antioxidants and proteins in ethylene-treated kiwifruits. Food Chem 107:640–648. https://doi.org/10.1016/j.foodchem.2007.08.070
Paulino N, Abreu SRL, Uto Y, Koyama D, Nagasawa H, Hori H, Dirsch VM, Vollmar AM, Scremin A, Bretz WA (2008) Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis. Eur J Pharmacol 587:296–301. https://doi.org/10.1016/j.ejphar.2008.02.067
Ramnath S, Venkataramegowda S, Singh C (2015) Chemical composition of bee Propolis collected from different regions in India by GCMS analysis. Int J Pharmacogn Phytochem 30:1319–1328
Romanha AJ, de Castro SL, Soeiro MDNC, Lannes-Vieira J, Ribeiro I, Talvani A, Bourdin B, Blum B, Olivieri B, Zani C, Spadafora C, Chiari E, Chatelain E, Chaves G, Calzada JE, Bustamante JM, Freitas-Junior LH, Romero LI, Bahia MT, Lotrowska M, Soares M, Andrade SG, Armstrong T, Degrave W, Andrade ZDA (2010) In vitro and in vivo experimental models for drug screening and development for Chagas disease. Mem Inst Oswaldo Cruz 105:233–238. https://doi.org/10.1590/S0074-02762010000200022
Roque ALR, Jansen AM (2014) Wild and synanthropic reservoirs of Leishmania species in the Americas. Int J Parasitol Parasites Wildl 3:251–262. https://doi.org/10.1016/j.ijppaw.2014.08.004
Roy S, Dutta D, Satyavarapu EM, Yadav PK, Mandal C, Kar S, Mandal C (2017) Mahanine exerts in vitro and in vivo antileishmanial activity by modulation of redox homeostasis. Sci Rep 7:1–16. https://doi.org/10.1038/s41598-017-03943-y
Schnitzler P, Neuner A, Nolkemper S, Zundel C, Nowack H, Sensch KH, Reichling J (2010) Antiviral activity and mode of action of propolis extracts and selected compounds. Phyther Res. https://doi.org/10.1002/ptr.2868
Sengupta S, Chowdhury S, BoseDasgupta S, Wright CW, Majumder HK (2011) Cryptolepine-induced cell death of Leishmania donovani Promastigotes is augmented by inhibition of autophagy. Mol Biol Int 2011:1–12. https://doi.org/10.4061/2011/187850
Shaha C (2006) Apoptosis in Leishmania species & its relevance to disease pathogenesis. Indian J Med Res 123:233–244
Sparg SG, Light ME, Van Staden J (2004) Biological activities and distribution of plant saponins. J Ethnopharmacol 94:219–243. https://doi.org/10.1016/j.jep.2004.05.016
Strober W (2001) Trypan blue exclusion test of cell viability. Curr Protoc Immunol. https://doi.org/10.1002/0471142735.ima03bs21
Sullivan DJ (2012) Cinchona alkaloids: quinine and quinidine. In: Staines H, Krishna S (eds) Treatment and prevention of malaria. Milestones in drug therapy. In: treatment and prevention of malaria: antimalarial drug chemistry, action and use, vol 41. Springer, Basel, pp 45–68. https://doi.org/10.1007/978-3-0346-0480-2_3
Tavakoli P, Shaddel M, Yakhchali M, Raoufi N, Shamsi H, Dastgheib M (2020) Antileishmanial effects of Propolis against Leishmania major in vitro and in vivo. Ann Mil Heal Sci Res 18:1–5. https://doi.org/10.5812/amh.100630
Tiwari P, Kumar B, Kaur M, Kaur G, Kaur H (2011) Phytochemical screening and extraction: a review. Int Pharm Sci 1:98–106
Tomiotto-Pellissier F, Alves DR, Miranda-Sapla MM, de Morais SM, Assolini JP, da Silva Bortoleti BT, Gonçalves MD, Cataneo AHD, Kian D, Madeira TB, Yamauchi LM, Nixdorf SL, Costa IN, Conchon-Costa I, Pavanelli WR (2018) Caryocar coriaceum extracts exert leishmanicidal effect acting in promastigote forms by apoptosis-like mechanism and intracellular amastigotes by Nrf2/HO-1/ferritin dependent response and iron depletion: Leishmanicidal effect of Caryocar coriaceum leaf ex. Biomed Pharmacother 98:662–672. https://doi.org/10.1016/j.biopha.2017.12.083
Zhou SH, Fang ZX, Lü Y, Chen JC, Liu DH, Ye XQ (2009) Phenolics and antioxidant properties of bayberry (Myrica rubra Sieb. Et Zucc.) pomace. Food Chem 112:394–399. https://doi.org/10.1016/j.foodchem.2008.05.104
Acknowledgments
The authors are thankful to the DST-CSIC, Post Graduate Institute of Medical Education and Research, Chandigarh for providing instrumentation facility. Sophisticated Analytical Instrument Facilities (SAIF)/ Central Instrumentation Laboratory (CIL), Panjab University, Chandigarh is acknowledged for GC-MS analysis.
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This work was financially supported by University Grants Commission, New Delhi under the grants, UGC-BSR JRF/SRF Fellowship [F.No.25–1/2014–15(BSR)/7–150/2007(BSR)], UGC-JRF/SRF Fellowship (3643/NET-DEC.2012) and Department of Science and Technology, GOI, New Delhi under the grant DST-FIST (SR/FST/LSI-545/2012(C)).
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DKG and PK carried out the experimental work and wrote the manuscript. DKG, PK, and SK designed the study. SK supervised the experimental study and reviewed the manuscript. All authors read and approved the final manuscript.
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Goyal, D.K., Keshav, P. & Kaur, S. Antiparasitic potential of Indian honey bee glue against strains of Leishmania donovani sensitive and resistant to synthetic antileishmanial. Biologia 76, 3841–3854 (2021). https://doi.org/10.1007/s11756-021-00897-5
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DOI: https://doi.org/10.1007/s11756-021-00897-5