African eggplant (<em>Solanum anguivi</em> Lam.) fruit with bioactive polyphenolic compounds exerts <em>in vitro</em> antioxidant properties and inhibits Ca<sup>2+</sup>-induced mitochondrial swelling
Objective
To evaluate the antioxidant and radical scavenging activities of Solanum anguivi fruit (SAG) and its possible effect on mitochondrial permeability transition pore as well as mitochondrial membrane potential (ΔΨm) isolated from rat liver.
Methods
Antioxidant activity of SAG was assayed by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, iron chelation and ability to inhibit lipid peroxidation in both liver and brain homogenate of rats. Also, the effect of SAG on mitochondrial membrane potential and mitochondrial swelling were determined. Identification and quantification of bioactive polyphenolics was done by HPLC-DAD.
Results
SAG exhibited potent and concentration dependent free radical-scavenging activity (IC50/DPPH=275.03±7.8 µg/mL). Reductive and iron chelation abilities also increase with increase in SAG concentration. SAG also inhibited peroxidation of cerebral and hepatic lipids subjected to iron oxidative assault. SAG protected against Ca (110 µmol/L)-induced mitochondrial swelling and maintained the ΔΨm. HPLC analysis revealed the presence of gallic acid [(17.54±0.04) mg/g], chlorogenic acid (21.90±0.02 mg/g), caffeic acid (16.64±0.01 mg/g), rutin [(14.71±0.03) mg/g] and quercetin [(7.39±0.05) mg/g].
Conclusions
These effects could be attributed to the bioactive polyphenolic compounds present in the extract. Our results suggest that SAG extract is a potential source of natural antioxidants that may be used not only in pharmaceutical and food industry but also in the treatment of diseases associated with oxidative stress.
Abstract
Objective
To evaluate the antioxidant and radical scavenging activities of Solanum anguivi fruit (SAG) and its possible effect on mitochondrial permeability transition pore as well as mitochondrial membrane potential (ΔΨm) isolated from rat liver.
Methods
Antioxidant activity of SAG was assayed by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, iron chelation and ability to inhibit lipid peroxidation in both liver and brain homogenate of rats. Also, the effect of SAG on mitochondrial membrane potential and mitochondrial swelling were determined. Identification and quantification of bioactive polyphenolics was done by HPLC-DAD.
Results
SAG exhibited potent and concentration dependent free radical-scavenging activity (IC50/DPPH=275.03±7.8 µg/mL). Reductive and iron chelation abilities also increase with increase in SAG concentration. SAG also inhibited peroxidation of cerebral and hepatic lipids subjected to iron oxidative assault. SAG protected against Ca (110 µmol/L)-induced mitochondrial swelling and maintained the ΔΨm. HPLC analysis revealed the presence of gallic acid [(17.54±0.04) mg/g], chlorogenic acid (21.90±0.02 mg/g), caffeic acid (16.64±0.01 mg/g), rutin [(14.71±0.03) mg/g] and quercetin [(7.39±0.05) mg/g].
Conclusions
These effects could be attributed to the bioactive polyphenolic compounds present in the extract. Our results suggest that SAG extract is a potential source of natural antioxidants that may be used not only in pharmaceutical and food industry but also in the treatment of diseases associated with oxidative stress.
Acknowledgments
The authors acknowledge the financial support of Educational Trust Fund, Nigeria and Doctoral fellowship in Professor Joao B.T. Rocha's Laboratory, Federal University of Santa Maria, Brazil. This research was funded by Educational Trust Fund, Nigeria with grant number ETF/ES/AST&D/AAU/AKUNGBA/1:2011.
Notes
Comments
Background
Mitochondrial permeability transition pore (MPTP) is activated due to oxidative stress. Antioxidants have been shown to prevent MPTP. The authors tried to established that polyphenolic compounds contained in fruit of SAG have antioxidant properties and are responsible for prevention of mitochondrial swelling which occur as a result of MPTP.
Research frontiers
The authors established that SAG fruit protected mitochondria membrane potential and prevented mitochondrial swelling and that the antioxidant capacity of SAG is responsible for this action. The HPLC analysis of SAG revealed the possible bioactive compounds responsible for these effects.
Related reports
This appears to be the first study on HPLC fingerprint, comprehensive antioxidants studies and effect of SAG fruit on mitochondrial membrane potential and mitochondrial swelling.
Innovations and breakthroughs
The antioxidant activity and HPLC fingerprint reveal the polyphenols from SAG fruit, prevention of mitochondria membrane permeability pore, and no effect on mitochondria membrane potential by fruit of SAG.
Applications
The study showed that the fruit of SAG is rich in antioxidants that can be used as food additives to prevent diseases associated with oxidative stress.
Peer review
This is a good study in which the authors established that SAG fruit has antioxidant activity, preventing mitochondria swelling and maintained mitochondria membrane potential. The results are very relevant because mitochondria swelling and a decrease of membrane potential has been implicated in many diseases associated with oxidative stress.
Footnotes
Foundation Project: Educational Trust Fund, Nigeria with grant number ETF/ES/AST&D/AAU/AKUNGBA/1:2011.
Conflict of interest statement: We declare that we have no conflict of interest.
References
- 1. Bolanos JP, Moro MA, Lizasoain I, Almeida AMitochondria and reactive oxygen and nitrogen species in neurological disorders and stroke: Therapeutic implications. Adv Drug Deliv Rev. 2009;61(14):1299–1315.[PubMed][Google Scholar]
- 2. Selivanov VA, Votyakova TV, Pivtoraiko VN, Zeak J, Sukhomlin T, Trucco M, et al. et al. Reactive oxygen species production by forward and reverse electron fluxes in the mitochondrial respiratory chain. Plos Comput Biol. 2011;7(3):1001115.
- 3. Valko M, Leibfritz D, Monco J, Cronin MT, Mazur M, Telser JFree radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 2007;39(1):44–84.[PubMed][Google Scholar]
- 4. Georgakilas AGOxidative stress, DNA damage and repair in carcinogenesis: Have we established a connection? Cancer Lett. 2012;327(1–2):3–4.[PubMed][Google Scholar]
- 5. Khansari N, Shakiba Y, Mahmoudi MChronic inflammation and oxidative stress as a major cause of age-related diseases and cancer. Recent Pat Inflamm Allergy Drug Discov. 2009;3(1):73–80.[PubMed][Google Scholar]
- 6. Khan MU, Cheema Y, Shahbaz AU, Ahokas RA, Sun Y, Gerling IC, et al. et al. Mitochondria play a central role in nonischemic cardiomyocyte necrosis: common to acute and chronic stressor states. Pflugers Arch. 2012;464(1):123–131.
- 7. Lindeque JZ, Levanets O, Louw R, van der Westhuizen FHThe involvement of metallothioneins in mitochondrial function and disease. Curr Protein Pept Sci. 2010;11(4):292–309.[PubMed][Google Scholar]
- 8. Perez-Alvarez S, Solesio ME, Manzanares J, Jordanand J, Galindo MFLactacystin requires reactive oxygen species and bax redistribution to induce mitochondria-mediated cell death. Br J Pharmacol. 2009;158(4):1121–1130.[Google Scholar]
- 9. Puntel RL, Roos DH, Folmer V, Nogueira CW, Galina A, Aschner M, et al. et al. Mitochondrial dysfunction induced by different organochalchogens is mediated by thiol oxidation and is not dependent of the classical mitochondrial permeability transition pore opening. Toxicol Sci. 2010;117(1):133–143.
- 10. Sivitz WI, Yorek MAMitochondrial dysfunction in diabetes: From molecular mechanisms to functional significance and therapeutic opportunities. Antioxid Redox Signal. 2010;12(4):537–577.[Google Scholar]
- 11. Corte CLD, Soares FAA, Aschner M, Rocha JBTDiphenyl diselenide prevents methylmercury-induced mitochondrial dysfunction in rat liver slices. Tetrahedron. 2012;68(5):10437–10443.[PubMed][Google Scholar]
- 12. Hung JC, Jong TK, En-Shyh LComparative antioxidant properties of water extracts from different parts of beefsteak plant (Perilla frutescens) J Food Drug Anal. 2009;17(6):489–496.[PubMed][Google Scholar]
- 13. Denton OA, Nwangburuka CCGenetic variability in eighteen cultivars of Solanum anguivi Lam. using principal component analysis (PCA) and single linkage cluster analysis (SLCA) Ann Biologic Res. 2011;2(4):62–67.[PubMed][Google Scholar]
- 14. Elekofehinti OO, Adanlawo IG, Saliu JA, Sodehinde SASaponins from Solanum anguivi fruits exhibit hypolipidemic potential in Rattus novergicus. Der Pharmacia Lett. 2012;4(3):811–814.[PubMed][Google Scholar]
- 15. Johnson M, Wesely EG, Selvan N, Chalini KComparative phytochemical and isoperoxidase studies on leaf and leaves derived callus of Solanum anguivi Lam. J Chem Pharm Res. 2010;2(4):899–906.[PubMed][Google Scholar]
- 16. Elekofehinti OO, Adanlawo IG, Fakoya A, Saliu JA, Sodehinde SAEffect of saponin from Solanum anguivi Lam. fruit on heart and kidney superoxide dismutase, catalase and malondialdehyde in rat. Curr Res J Biol Sci. 2012;4(4):530–533.[PubMed][Google Scholar]
- 17. Elekofehinti OO, Adanlawo IG, Komolafe K, Ejelonu OCSaponins from Solanum anguivi fruits exhibit antioxidant potential in Wistar rats. Ann Biol Res. 2012;3(7):3212–3217.[PubMed][Google Scholar]
- 18. Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OGDetermination of the total phenolic, flavonoid and praline contents in Burkina fasan honey, as well as their radical scavenging activity. Food Chem. 2005;91:571–577.[PubMed][Google Scholar]
- 19. Kamdem JP, Stefanello ST, Boligon AA, Wagner C, Kade IJ, Pereira RP, et al. et al. In vitro antioxidant activity of stem bark of Trichilia catigua Adr. Juss (Meliaceae) Acta Pharm. 2012;62:371–382.[PubMed]
- 20. Shanab SM, Mostafa SS, Shalaby EA, Mahmoud GIAqueous extracts of microalgae exhibit antioxidant and anticancer activities. Asian Pac J Trop Biomed. 2012;2(8):608–615.[Google Scholar]
- 21. Puntel RL, Nogueira CW, Rocha JBKrebs cycle intermediates modulate thiobarbituric reactive species (TBARS) production in rat brain in vitro. Neurochem Res. 2005;30(2):225–235.[PubMed][Google Scholar]
- 22. Kumar RS, Rajkapoor B, Perumal PAntioxidant activities of Indigofera cassioides Rottl. Ex. DC. using various in vitro assay models. Asian Pac J Trop Biomed. 2012;2(4):256–261.[Google Scholar]
- 23. Ohkawa H, Ohishi N, Yagi KAssay for lipid peroxide in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95(2):351–358.[PubMed][Google Scholar]
- 24. Puntel RL, Roos DH, Folmer V, Nogueira CW, Galina A, Aschner M, et al. et al. Mitochondrial dysfunction induced by different organochalchogens is mediated by thiol oxidation and is not dependent on the classical mitochondrial permeability transition pore opening. Toxicol Sci. 2010;117(1):133–143.
- 25. Votyakova TV, Reynolds IJCa-induced permeabilization promotes free radical release from rat brain mitochondria with partially inhibited complex I. J Neurochem. 2005;93(3):526–537.[PubMed][Google Scholar]
- 26. Bradford MMA rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1979;72:248.[PubMed][Google Scholar]
- 27. Sabir SM, Ahmad SD, Hamid A, Khan MQ, Athayde ML, Santos DB, et al. et al. Antioxidant and hepatoprotective activity of ethanolic extract of leaves of Solidago microglossa containing polyphenolic compounds. Food Chem. 2012;131(3):741–747.[PubMed]
- 28. Zahin M, Aqil F, Ahmad IThe in vitro antioxidant activity and total phenolic content of four indian medicinal plants. Int J Pharm Pharm Sci. 2009;1:88–95.[PubMed][Google Scholar]
- 29. Gulcin I, Bursal E, Sehitoglu MH, Bilsel M, Goren ACPolyphenol contents and antioxidant activity of lyophilized aqueous extract of propolis from Erzurum, Turkey. Food Chemi Toxicol. 2010;48(8–9):2227–2238.[PubMed][Google Scholar]
- 30. Thenmozhi A, Nagalakshmi K, Rao UQualitative analysis of phytochemicals, and comparative superoxide radical scavenging along with reducing potency of Solanum nigrum using various solvent extracts. Int J Green Pharm. 2011;5(4):318–324.[PubMed][Google Scholar]
- 31. Costa P, Goncalves S, Valentao P, Andrade PB, Coelho N, Romano A. Thymus lotocephalus wild plants and in vitro cultures produce different profiles of phenolic compounds with antioxidant activity. Food Chem. 2012;135:1253–1260.[PubMed]
- 32. Ikeda H, Kimura Y, Masaki M, Iwahashi HCaffeic acid inhibit the formation of 1-hydroxyethyl radical in the reaction mixture of rat liver microsomes with ethanol partly through its metal chelating activity. J Cli Biochem Nutr. 2011;48(3):187–193.[Google Scholar]
- 33. Elekofehinti OO, Kade IJAqueous extract of African egg plant inhibit Fe and SNP induced lipid peroxidation in rat's brain- In vitro. Der Pharma Lett. 2012;4(5):1352–1359.[PubMed][Google Scholar]
- 34. Omololu PA, Rocha JB, Kade IJAttachment of rhamnosyl glucoside on quercetin confers potent iron-chelating ability on its antioxidant properties. Exp Toxicol Pathol. 2011;63(3):249–255.[PubMed][Google Scholar]
- 35. Kaurinovic B, Popovic M, Vlaisavljevic S, Schwartsova H, Vojinovic-Miloradov MAntioxidant profile of Trifolium pratense L. Molecules. 2012;17(9):11156–11172.[Google Scholar]
- 36. Nair SS, Nithyakala CM, Rozario RV, Jennifer J, Somashekharaiah BVBiochemical characterization of selected plant species and investigation of phytochemicals for in-vitro antioxidant activity. Int J Pharmacogn Phytochem Res. 2012;4(3):127–133.[PubMed][Google Scholar]
- 37. Mehta A, Prabhakar M, Kumar P, Deshmukh R, Sharma PLExcitotoxicity: Bridge to various triggers in neurodegenerative disorders. Eur J Pharmacol. 2013;698(1–3):6–18.[PubMed][Google Scholar]
- 38. Costa RO, Ferreiro E, Oliveira CR, Pereira CMFInhibition of mitochondrial cytochrome c oxidase potentiates Aβ-induced ER stress and cell death in cortical neurons. Mol Cellular Neurosci. 2013;52:1–8.[PubMed][Google Scholar]
- 39. Eckmann J, Eckert SH, Leuner K, Muller WE, Eckert GPMitochondria: mitochondrial membranes in brain ageing and neurodegeneration. Int J Biochem Cell Biol. 2013;45(1):76–80.[PubMed][Google Scholar]
- 40. Javadov S, Karmazyn M, Escobales NMitochondrial permeability transition pore opening as a promising therapeutic target in cardiac diseases. J Pharmacol Exp Ther. 2007;330:670–678.[PubMed][Google Scholar]
- 41. Norenberg MD, Rao KVThe mitochondrial permeability transition in neurologic disease. Neurochem Int. 2007;50(7–8):983–997.[Google Scholar]
- 42. Tarba CRelationship between membrane potential, calcium fluxes and matrix swelling in rat liver mitochondria; effect of ethanol feeding. Ann Romanian Soc Cell Biol. 2009;14:9–27.[PubMed][Google Scholar]
- 43. Ge R, Ma WH, Li YL, Li QSApoptosis induced neurotoxicity of Di-n-butyl-di-(4-chlorobenzohydroxamato) Tin (IV) via mitochondria-mediated pathway in PC12 cells. Toxicol In Vitro. 2013;27(1):92–102.[PubMed][Google Scholar]
- 44. Selimovic D, Porzig BB, El-Khattouti A, Badura HE, Ahmad M, Ghanjati F, et al. et al. Bortezomib/proteasome inhibitor triggers both apoptosis and autophagy-dependent pathways in melanoma cells. Cell Signal. 2013;25(1):308–318.[PubMed]
- 45. Greetham D, Kritsiligkou P, Watkins RH, Carter Z, Parkin J, Grant CMOxidation of the yeast mitochondrial thioredoxin promotes cell death. Antioxid Redox Signal. 2013;18:375–385.[Google Scholar]
- 46. Sloan RC, Moukdar F, Frasier CR, Patel HD, Bostian PA, Lust RM, et al. et al. Mitochondrial permeability transition in the diabetic heart: Contributions of thiol redox state and mitochondrial calcium to augmented reperfusion injury. J Mol Cell Cardiol. 2012;52(5):1009–1018.[PubMed]
- 47. Petronilli V, Sileikyte J, Zulian A, Dabbeni-Sala F, Jori G, Gobbo S, et al. et al. Switch from inhibition to activation of the mitochondrial permeability transition during hematoporphyrin-mediated photooxidative stress. Unmasking pore-regulating external thiols. Biochim et Biophys Acta. 2009;1787:897–904.[PubMed]
- 48. Halestrap AP, Clarke SJ, Javadov SAMitochondrial permeability transition pore opening during myocardial reperfusion-a target for cardioprotection. Cardiovasc Res. 2004;61(3):372–385.[PubMed][Google Scholar]
- 49. Wu PF, Zhang Z, Wang F, Chen JGNatural compounds from traditional medicinal herbs in the treatment of cerebral ischemia/reperfusion injury. Acta Pharmacol Sin. 2010;31(12):1523–1531.[Google Scholar]
- 50. Javadov S, Karmazyn MMitochondrial permeability transition pore opening as an endpoint to initiate cell death and as a putative target for cardioprotection. Cell Physiol Biochem. 2007;20(1–4):1–22.[PubMed][Google Scholar]
- 51. Madesh M, Hajnoczky GVDAC-dependent permeabilization of the outer mitochondrial membrane by superoxide induces rapid and massive cytochrome c release. J Cell Biol. 2001;155(6):1003–1016.[Google Scholar]