Toxic Effects of Unripe Carica papaya (Linn) Fruit Extract on Healthy Rat Liver Mitochondria
Abstract
The opening of the mitochondrial Permeability Transition (mPT) pore proceeds the activation of programmed cell death (apoptosis) and its functional status serves as marker of mitochondrial health. Unripe fruits of Carica papaya are used in the traditional treatment of several diseases. There is paucity of information on the level of safety of the consumption of the plant. The effects of crude Methanol Extract of Carica papaya (MECP) on the status of the mPT pore in healthy rat liver was investigated in this study. Mitochondrial FOF1 ATPase activity, mitochondrial permeability transition and mitochondrial lipid peroxidation as well as the release of cytochrome c were evaluated spectrophotometrically using standard methods. The MECP activated mPT pore opening in the absence of calcium in a concentration-dependent fashion. Specifically, induction folds of 3.1, 6.0, 9.1, 11.9 and 14.3 were recorded at 20, 60,100, 140and 180 µg/ml, respectively. In addition, MECP potentiated calcium-induced pore opening of the mPT pore in a concentration-dependent style by 22.5, 24.1, 25.0, 25.1 and 25.5 folds, respectively at 20, 60, 100, 140 and 180 µg/mL. Furthermore, mitochondrial ATPase activity was significantly (p < 0.001) stimulated at pH (7.4) while the extent of cytochrome c release increased by 5 and 7 folds respectively at the highest concentrations tested. Interestingly, Fe2+-induced mitochondrial lipid peroxidation was inhibited by varying concentrations of MECP. Specifically, significant (p < 0.001) reduction in levels of mitochondrial lipid peroxides were observed at 50, 100, 200, 300, 600 µg/ml MECP by 10, 22, 53, 74, 112 %, respectively. These findings indicate that unripe Carica papaya fruit extract contains bioactive compounds that cause mitochondrial injury via activation of the mitochondrial permeability transition pore opening in healthy liver cells. Hence, its use in the management of diseases should be approached with caution.
Rakotoarivelo NH, Rakotoarivony F, Ramarosandratana AV, Jeannoda VH, Kuhlman AR et al. Medicinal plants used to treat the most frequent diseases encountered in Ambalabe rural community, Eastern Madagascar. J Ethnobiol Ethnomed. 2015;11:68.
Petran M, Dragos D, Gilca M. Historical ethnobotanical review of medicinal plants used to treat children diseases in Romania (1860s–1970s). J Ethnobiol Ethnomed 2020;16:15.
Amadi CN, Orisakwe OE. Herb-Induced Liver Injuries in Developing Nations: An Update. Toxics 2018;6:24.
Lin NH, Yang HW, Su YJ, Chang CW. Herb induced liver injury after using herbal medicine: A systemic review and case-control study. Medicine (Baltimore) 2019;98:e14992.
Santana LF, Inada AC, Espirito Santo BLSD, Santana LF, Inada AC et al. Nutraceutical Potential of Carica papaya in Metabolic Syndrome. Nutrients 2019;11:1608.
Benarba B, Pandiella A. Medicinal Plants as Sources of Active Molecules against COVID-19. Front Pharmacol 2020;11:1189.
Singh S, Kumar S, Mathan, S, Tomar MS, Singh RK et al. Therapeutic application of Carica papaya leaf extract in the management of human diseases. DARU J Pharm Sci 2020;28:735-744.
Vij T, Prashar Y. A review on medicinal properties of Carica papaya Linn. Asian Pac J Trop Dis 2015;5:16.
Longdet I, Adoga E. Effect of methanolic leaf extract of Carica papaya on plasmodium berghei infection in albino mice. Eur J Med Plants 2017;20:1-7.
Saran P, Choudhary R. Drug bioavailability and traditional medicaments of commercially available papaya: A review. Afr J Agric Res 2013;8:3216-3223.
Shouket H, Ameen I, Tursunov O, Kholikova K, Pirimov O et al, Study on industrial applications of papain: A succinct review. IOP Conf. Ser. Earth Environ Sci 2020;614:012171.
Okon W, Ogri I, Igile GO, Atangwho I. Nutritional quality of raw and processed unripe Carica papaya fruit pulp and its contribution to dietary diversity and food security in some peasant communities in Nigeria. Int J Biol Chem Sci 2017;11:1000-1011.
Jarisarapurin W, Sanrattana W, Chularojmontri L, Kunchana K, Wattanapitayakul S. Antioxidant Properties of Unripe Carica papaya Fruit Extract and Its Protective Effects against Endothelial Oxidative Stress. Evid Based Complement Altern Med 2019;1-15.
Karemore MN, Avari JG. Herbal medicines used during pregnancy, childbirth and postpartum care. Int J Pharm Sci Res 2017;8:5326-5335.
Pandey S, Cabot P, Shaw P, Hewavitharana A. Anti-inflammatory and immunomodulatory properties of Carica papaya. J Immuno Toxicol 2016;13:590-602.
Norahmad NA, Mohd Abd Razak MR, Mohmad Misnan N. Md Jelas NH, Sastu UR et al. Effect of freeze-dried Carica papaya leaf juice on inflammatory cytokines production during dengue virus infection in AG129 mice. BMC Complement Altern Med 2019;19:44.
Kumari S, Kumar R, Singh V. Contraceptive effects of aqueous extract of Carica papaya (Linn.) seed on seminal profile of swiss albino mice. Int J Sci Res 2015;6:1963-1965.
Nwaehujor OC, Ode OJ, Ekwere MR, Udegbunam RI. Anti-fertility effects of fractions from Carica papaya (Pawpaw) Linn. methanol root extract in male Wistar rats, Arab J Chem 2019;12:1563-1568.
Soib HH, Ismail HF, Husin F, Abu Bakar MH, Yaakob H et al. Bioassay-guided different extraction techniques of Carica papaya (Linn.) Leaves on in vitro wound-healing activities. Molecules 2020;25:517.
Nafiu AB, Rahman MT. Anti-inflammatory and antioxidant properties of unripe papaya extract in an excision wound model. Pharm Biol 2014;53:662-671.
Farina V, Tinebra I, Perrone A, Sortino G, Palazzolo E et al. Physicochemical, nutraceutical and sensory traits of six papaya (Carica papaya L.) cultivars grown in greenhouse conditions in the mediterranean climate. Agronomy 2020;10:501.
Aruoma O, Somanah J, Bourdon E, Rondeau P, Bahorun T. Diabetes as a risk factor to cancer: functional role of fermented papaya preparation as phytonutraceutical adjunct in the treatment of diabetes and cancer. Mutat Res 2014;768:60-68.
Sagnia B, Fedeli D, Casetti R, Montesano C, Falcioni G et al. Antioxidant and anti- inflammatory activities of extracts from Cassia alata, Eleusine indica, Eremomastax speciosa, Carica papaya and Polyscias fulva medicinal plants collected in Cameroon. PLoS One 2014:9:1-10.
Oyebode O, Odejide T, Kukoyi A, Adebisi L, Olorunsogo O. Effects of Different Fractions of Calliandra portoricensis Root Bark on Isolated Rat Liver Mitochondrial Membrane Permeability Transition Pore. Afr J Med Med Sci 2012;41:399-409.
Olanlokun J, Oyebode O, Olorunsogo O. Effects of vacuum liquid chromatography (chloroform) fraction of the stem bark of Alstonia boonei on mitochondrial membrane permeability transition pore. J Basic Clin Pharm 2017;84:221-225.
Olowofolahan AO, Oyebode OT, Olorunsogo OO. GCMS analysis of partially purified chloroform sub fractions of methanol extract of drymaria cordata (Linn) and their effects on mitochondrial membrane permeability transition pore. Arch Bas App Med 2020;8:3-11.
Halestrap A, Clarke S, Javadov S. Mitochondrial permeability transition pore opening during myocardial reperfusion- a target for cardioprotection. Cardiovasc Res 2004;61:372-385.
Halestrap A, Richardson A. The mitochondrial permeability transition: a current perspective on its identity and role in ischaemia/ reperfusion injury. J Mol Cell Cardiol 2015;78:129-141.
Oyebode O, Akinyelu J, Oamen E, Olorunsogo O. Methanol fraction of Calliandra portoricensis root bark activates caspases via alteration in mitochondrial viability in vivo. J. HerbMed Pharmacol 2018;7:251-258.
Crompton M. The mitochondrial permeability transition pore and its role in cell death. Biochem J 1999;341:233-249.
Bauer T, Murphy E. Role of mitochondrial calcium and the permeability transition pore in regulating cell death. Circ Res 2020;126:280-293.
Afzan A, Abdullah NR, Halim SZ, Rashid BA, Semail RH et al. Repeated Dose 28-Days Oral Toxicity Study of Carica papaya L. leaf extract in sprague dawley rats. Molecules 2012;17:4326-4342.
Kanadi MA, Alhassan A, Ngwen A, Yaradua A. Nasir A et al. Acute toxicity studies and phytochemical constituents of different solvents extracts of carica papaya Seeds. Asian Journal of Research in Botany 2019;2:1-9.
Kharisma Y, Rachmawati M, Indriyanti RA, Respati T. Unripe Papaya (Carica papaya L.) Aqueous Extract (UPAE) for the acute toxicity test on fatty liver changes. J Phys Conf Ser 2020; 1469:012013.
Mohammad I. Gold nanoparticle: An efficient carrier for MCP I of Carica papaya seeds extract as an innovative male contraceptive in albino rats. J Drug Deliv Sci Technol 2019;52: 942-956.
Johnson D, Lardy H. Isolation of liver or kidney mitochondria. Methods Enzymol 1967;10:94-96.
Olorunsogo O, Bababunmi E, Bassir O. Uncoupling effects of N-phosphonomethyl glycine on rat liver mitochondria. Biochem Pharm 1979;27:925-927.
Lapidus R, Sokolove P. Inhibition by Spermine of the inner membrane permeability transtion of isolated rat heart mitochondria. FEBS Lett 1992;3:314-318.
Lowry O, Rosenbrough N, Farr A, Randall R, Protein measurement with protein phenol reagent. J Biol Chem 1951;193:265-275.
Olorunsogo O, Malomo S. Sensitivity of oligomycin-inhibited respiration of isolated rat liver mitochondria to perfluidone, a fluorinated arylalkylsulfonamide. Toxicology 1985;35:231-240.
Bassir O. Handbook of Practical Biochemistry. Ibadan University press, Ibadan 1963; p 13.
Ruberto G, Baratta M, Deans S, Dorman H. Antioxidant and antimicrobial activity of Foeniculum vulgare and Crithmum maritimum essential oils. Planta Med 2000;66:687-693.
Appaix F, Minatchy M, Riva-Lavieille C, Olivares J, Antonsson B et al. Rapid spectrophotometric method for quantitation of cytochrome c release from isolated mitochondria or permeabilized cells revisited. Biochim Biophys Acta 2000;1457:175-181.
Naoi M, Wu Y, Shamoto-Nagai M, Maruyama W. Mitochondria in neuroprotection by phytochemicals: bioactive polyphenols modulate mitochondrial apoptosis system, function and structure. Int J Mol Sci 2019;20:2451.
Bernardi P. Why F-ATP synthase remains a strong candidate as the mitochondrial permeability transition pore. Front Physiol 2018;9:1543.
Gerle C. Mitochondrial F-ATP synthase as the permeability transition pore. Pharmacol Res 2020;160:105081.
Kalani K, Yan SF, Yan SS. Mitochondrial permeability transition pore: a potential drug target for neurodegeneration. Drug Discov Today 2018;23:1983-1989.
Baines C, Gutierrez-Aguilar M. The still uncertain identity of the channel-forming unit(s) of the mitochondrial permeability transition pore. Cell Calcium 2018;73:121-130.
Chinopoulos C. Mitochondrial permeability transition pore: Back to the drawing board. Neurochem Int 2018;117:49-54.
Bhosale G, Duchen MR. Investigating the mitochondrial permeability transition pore in disease phenotypes and drug screening. Curr Protoc Pharmacol 2019;85:e59.
Juárez-Rojop IE, Tovilla-Zárate CA, Aguilar-Domínguez DE, Roa-de la Fuente LF, Lobato-García CE, et al. Phytochemical screening and hypoglycemic activity of Carica papaya leaf in streptozotocin-induced diabetic rats. Rev Bras Farmacogn 2014;24:341-347.
Seidlmayer L, Gomez-Garcia M, Blatter L, Pavlov E, Dedkova E. Inorganic polyphosphate is a potent activator of the mitochondrial permeability transition pore in cardiac myocytes. J Gen Physiol 2012;139:321-331.
Files | ||
Issue | Vol 6, No 2, 2021 | |
Section | Research Article(s) | |
DOI | https://doi.org/10.18502/tim.v6i2.6789 | |
Keywords | ||
Mitochondria Carica papaya Permeability transition FOF1 ATPase activity |
Rights and permissions | |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |