Some Solvent Fractions of the Fruits of Xylopia aethiopica Enhance Mitochondrial-Mediated Apoptosis in Rat Liver

  • Olufunso Olabode Olorunsogo Laboratories for Biomembrane Research and Biotechnology, Biochemistry Department
  • Oluwatosin Clara Obigade


The mitochondrial membrane Permeability Transition (mPT) pore has emerged as a promising target for drug development in diseases where there is dysregulation of apoptosis. In this study, the effects of various fractions of the methanol extract of Xylopia aethiopica (XA), a potent medicinal plant, were investigated on mitochondrial-mediated apoptosis. The methanol extract was partitioned in succession between dichloromethane, ethylacetate and methanol to obtain Dichloromethane (DFXA), Ethylacetate (EFXA) and Methanol (MFXA) fractions of the fruits. The effects of DFXA, the most potent fraction, on mPT pore, mitochondrial ATPase (mATPase), lipid peroxidation, DNA fragmentation, cytochrome c release and caspases 9 and 3 activities were estimated. Varying concentrations of DFXA (20, 60, 100, 140 and 180 μg/ml) significantly induced pore opening in the absence of calcium by 9.92, 12.36, 13.75, 14.92 and 15.47 folds, respectively. Similarly, Ca 2+-induced mPT pore opening was further enhanced by 8.3, 9.14, 10.5, 18.53 and 20.5 folds and mATPase activity was significantly elevated by these concentrations of DFXA. In contract, EFXA and MFXA did not have any significant effect at lower concentrations but induced pore opening at 180μg/ml by 1.69 and 6.4 folds, respectively. In vivo, DFXA caused the induction of mPT pore in the absence of calcium, activation of the activities of caspases 9 and 3, significant DNA fragmentation in a dose-dependent manner and also ameliorated ferrous-induced mitochondrial membrane lipid peroxidation. These findings reveal that the dichloromethane fraction of Xylopia aethjopica contains a bioactive agent capable of inducing mitochondrial-mediated apoptosis and may be useful for drug development in diseases where apoptosis is compromised


Acehan D, Jiang X, Morgan DG, Heuser JE, Wang X, Akey CW (2002). Three-dimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation. Mol Cell 9:423-32.
Adam-Vizi V. and Seregi A. (1982). Receptor independent stimulatory effect of noradrenaline on Na, K-ATPase in rat brain homogenate: Role of lipid peroxiadation. Biochem. Pharmacol. 31, 2231-2236.
Adaramoye OA, Sarkar J, Singh N, Meena S, Changkija B, Yadav PP, Kanojiya S and Sinha S (2011). Antiproliferative Action of Xylopia aethiopica Fruit Extract on Human Cervical Cancer Cells. Phytother. Res. DOI: 10.1002/ptr.3551.
Adedosu OT, Aejoke TT, Salako OO, Olorunsogo OO. (2012). Effects of extract of the leaves of Brysocarpus coccineus on rat liver mitochondrial membrane permeability transition (MMPT) pore. Afr J Med Med Sci. 41 Suppl:125-32.
Adisa RA, Olorunsogo OO. (2013). Robustaside B and para hydroxyphenol: Phenolic and antioxidant compounds purified from Cnestis ferruginea D.C induced membrane permeability transition in rat liver mitochondria. Molecular Medicine Reports 8.5 1493-1498.
Adisa, R., M. Choudhary, E. Adewoye and O. Olorunsogo, (2010). Hypoglycaemic and biochemical properties of Cnestis ferruginea. Afr. J. Tradit. Complement Altern Med., 7: 185-194.
Anyasor GN, Ajayi E.I., Saliu J.A., Ajagbona O., and Olrunsogo O.O. (2009). Artensunate opens mitochondrial membrane permeability transition (MMPT) pore. Annals of Trop. Med. Publ. Health 2, 37-41.
Appaix, F., Minatchy, M., Riva-Lavieille, C., Olivaires, J., Antonnson, B. and Saks, V. A. (2000). Rapid Spectrophotometric method for quantitation of cyctochrome c release from isolated mitochondria or permeabilized cells revisited. Biochimica et Biophysica Acta 1457: 175-181.
Bassir,O. (1963). Handbook of Practical Biochemistry. Ibadan University Press, Ibadan, Nigeria p 31
Belzacq AS, El Hamel C, Vieira HL, Cohen I, Haouzi D, Metivier D, et al, (2001). Adenine nucleotide translocator mediates the mitochondrial membrane permeabilization induced by Ionidamine, arsenite and CD437. Oncogene 20, 7579-7587.
Bernardi P, (2018). Why F-ATP synthase remains a strong candidate as the mitochondrial permeability transition pore. Frontiers in Physol. 9:(1543) 1-4
Bounival J, Quessy P, Martinol MG. (2009). Protective effects of resveratrol and quercetin against MPP+-induced oxidative stress act by modulating markers of apoptotic death in dopaminergic neurons. Cell. Mol. Neurobiol. 29, 1169-1180.
Briston T, Selwood AL, Szabadkai G and Duchen MR (2018). Mitochondrial permeability transition: a molecular lesion with multiple drug targets. Trends in pharmacological sciences 1570.
Del-Rio, A., B.G. Obdulio, J. Castillo, F.R. Main and A. Ortuno, (1997). Uses and properties of Citrus flavonoids. J. Agric. Food Chem., 45: 4505-15.
Ezekwesili, C.N., Nwodo, O.F.C., Eneh, F.U., Ogbunugafor, H.A., (2010). Investigation of the chemical composition and biological activity of Xylopia aethiopica Dunal (Annonacae). Afr. J. Biotech., 9(43): 7352-7356.
Fadeel B, Gleiss B, Hogstrand K, Chandra J, Wiedmer T, Sims PJ, Henter JI, Orrenius S and Samali A (1999). “Phosphatidylserine exposure during apoptosis is a cell-type-specific event and does not correlate with plasma membrane phospholipid scramblase expression. “Biochem Biophs Res Commun 266(2): 504-11.
Fleischer TC, Mensah ML, Mensat AY, Komiuga SY, Gbedemg SY, Skaltsa H, (2008). Antimicrobial activity of essential oils of Xylopia aethiopica. Afr J Trad Complement Altern Med, 4:391-393.
Fulda S, Sieverts H, Friesen C, Herr I, Debatin KM. (1997). The CD95 (APO-1/Fas) system mediayes drug induced apoptosis in neuroblastoma cells. Cancer Res. 57: 3823–3829.
Fulda S. (2010). Modulation of apoptosis by natural products for cancer therapy. Planta Med. 76:1075-9.
Huang L, Chen C, Zhang X, Li X, Chen Z, Yang C, Lianf X, Zhu G, Xu Z. neuroprotective effect of curcumin against ceredral ischemia-reperfusion via mediating autophagy and inflammation. J. Mol. Neurosci. 2018,64, 129-139.
Javadov S and Karmazyn M (2007) Mitochondrial permeability transition pore opening as an endpoint to initiate cell death and as a putative target for cardioprotection. Cell Physiol Biochem 20:1–22.
John-Dewole, O.O., Agunbiade, S.O., Alao O.O., and Arojojoye O.A. (2012). Phytochemical and antimicrobial studies of extract of the fruit of Xylopia aethiopica for medicinal importance. Journal of Biotechnology and Pharmaceutical Research Vol. 3(6), pp. 118-122.
Johnson D, Lardy H (1967). Isolation of liver or kidney mitochondria. Methods Enzymol 10:94-6.
Kerr JF, Wyllie AH, Currie AR (1972). “Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics”. Br J Cancer 26(4): 239-57.
Lapidus, R.G. and Sokolove, P.M. (1993). Spermine inhibition of permeability transition of isolated rat liver mitochondria; An investigation of mechanism. ArchBiochem Biophys 306(1):246-253.
Lardy HA, and Wellman H (1953). The catalytic effect of 2,4 dinitrophenol on adenosinetriphosphate hydrolysis by cell particles and soluble enzymes J. Biol. Chem. 201:351-370.
Lopez J and Tait C (2015). Mitochondrial apoptosis: killing cancer using the enemy within. British Journal of Cancer. 112(6).10.1038/bjc.2015.85.
Lowry, O.H., Rosenbrough, N.J., Farr, A.L. and Randall, R.J. (1951). Protein measurement with Folin phenol reagent. J Biol Chem193:265-275.
Marchetti P, Zamzami N, Joseph B, Schraen-Maschke S, Me´reau-Richard C, Costantini P, Me´tivier D, Susin SA, Kroemer G, and Formstecher P (1999). The Novel Retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphtalene Carboxylic Acid can Trigger Apoptosis through a Mitochondrial Pathway Independent of the Nucleus. Cancer res 59: 6257–6266.
Meier P, Finch A, and Evan G (2000). Apoptosis in development.’Nature 407(6805):796-801.
Odewusi AF, Opeyemi MO, Olayemi FO, Emikpe B, Ehigie LO, Adisa RA and Olorunsogo OO (2010). Effects of the leaf decoction of Momordica charantia (bitter melon) on Mitochondrial Membrane Permeability Transition Pore (MMPTP) and fertility in normal male albino rats. Afr.J.Med.Sci. 39, Suppl. 47-59.
Ogunkunle, A.T.S. and T.A. Ladejobi, (2006). Ethnobotanical and phytochemical studies on some species of Senna in Nigeria. Afr. J. Biotechnol., 5(21): 2020-2023.
Olorunsogo O.O and Malomo S.O (1985). Sensitivity of Oligomycin inhibited respiration of isolated rat liver mitochondriato perfluidone, a fluorinated arylalkylsulfonamide. Toxicology, 35 (3):231-40.
Olorunsogo, O.O., Bababunmi, E.A. and Bassir, O. (1979). Uncoupling effect of N-(phosphonomethyl)glycine on at liver mitochondria. Biochem. Pharm. 27:925- 927.
Oyebode OT, Odejide TT, Kukoyi AJ, Adebisi LA, Olorunsogo OO (2012). Effects of different fractions of Calliandra portoricensis root bark on isolated rat liver mitochondrial membrane permeability transition pore. Afr J Med Med Sci. 41:399-409.
Oyebode TO, Adebusuyi OT, Akintimehin SE, Olorunsogo OO (2017). Modulation of cytochrome C release and opening of mitochondrial permeability transition pore by Calliandra portoricensis (Benth) root bark methanol extract. European J Med Plants. 20(1):1-14. doi: 10.9734/ EJMP/2017/35211.
Ruberto G, Baratta M, Deans S, Dorman H. (2000). Antioxidant and antimicrobial activity of Foeniculum vulgare and Crithmum maritimum essential oils. Planta Med 66: 687 -693.
Salvesen GS and Renatus M (2002). Apoptosome: the seven-spoked death machine. “Dev Cell 2(3): 256-7
Selzer E, Thallinger C, Hoeller C, Oberkleiner P, Wacheck V, Pehamberger H, Jansen B (2002). Betulinic acid-induced Mcl-1 expression in human melanoma-mode of action and functional significance. Mol Med 8:877-884.
Vashney,R. and Kale, R.K. (1990). Effects of calmodulin Antagonism on radiation induced lipid peroxidation in microsomes. Int. J Rad.Biol. 58:733-743.
Wu B., Ootani A., Iwakiri R., Sakata Y., Fujise T., Amemori S., Yokoyama F., Tsunada S., Toda S. and Fujimoto K. (2006). T cell deficiency leads to liver carcinogenesis in azoxymethane-treated rats. Exp Biol Med 231:91-98.
Wyllie AH (1980). Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 284:555-556
Yusuf AZ, Zakir A, Shemau Z, Abdullahi M, Halima SA (2014). Phytochemical analysis of the methanol leaves extract of Paullinia pinnata linn. J. Pharmacogn. Phytother. 6(2):10-16.
Nutrition/Natural Product and Drug Development