Abstrakt
Background: Ocimum gratissimum Linn. leaves have been shown to be useful in arthritis as well as related inflammatory oxidative conditions. The study investigated the cytotoxicity, membrane stabilizing and antiarthritic effects of sequential methanol extract of the leaves.
Methods: Powdered leaves were sequentially extracted with n-hexane, chloroform and methanol to obtain a resultant methanol extract (MEOg). The extract was tested for cytotoxicity using Chinese harmster ovary cell (CHO-k1) and murine macrophages (RAW 264.7) cell lines. Membrane stabilizing effect was tested by heatinduced erythrocytes haemolysis assay. Antiarthritic properties were tested by egg-albumin, formalin-induced inflammation and carrageenan/kaolin induced monoarthritis models in rats. Thereafter, biomarkers of oxidative stress were evaluated using standard biochemical assays.
Results: The extract did not reduce viability of CHO-k1 and RAW 264.7 cells. MEOg showed significant (p <0.05) membrane stabilizing properties. MEOg at 100, 200, and 400 mg/kg dose-dependently reduced egg albumin-induced paw oedema by 24.0, 32.2, and 37.8%, respectively. Similarly, MEOg significantly (p < 0.05) decreased paw thickness compared to control in formalin-induced arthritis in rats. Furthermore, MEOg dose-dependently showed anti-inflammatory activity which was evident with decrease in paw and knee swelling, and decreased inflammatory pain in carrageenan/kaolin-induced monoarthritis in rats. The treatment decreased plasma TBARS and NO, and increased GSH and SOD levels.
Conclusion: These findings support the ethno-pharmacological use of Ocimum gratissimum leaves for ameliorating inflammation and pain of arthritis.
Keywords: Cytotoxicity, Anti-arthritic, Ocimum gratissimum, carrageenan, kaolin
Résumé
Contexte: Ila été montré que lesfeuilles d’ocimumgratissimumLinn. sont utiles dans l’arthrite ainsi que dans les conditions oxydatives inflammatoires associées.L’étude a étudié la propriété cytotoxique, la stabilisation membranaire et les effets antiarthritiques de l’extrait séquentiel de méthanol des feuilles.
Méthodes: Les feuilles en poudre ont été extraites séquentiellement avec du n-hexane, du chloroforme et du méthanol pour obtenir un extrait méthanoïque résultant (MEOg).L’extrait a été testé pour sa propriété cytotoxique en utilisant deslignées cellulaires d’ovaire harmonique chinois(CHO-k1) et de macrophages murins (RAW 264.7).L’effet de stabilisation de la membrane a été testé par untest d’hémolyse des érythrocytes induit par la chaleur. Les propriétésantiarthritiquesont été testées par desmodèles d’albumine d’œuf, de l’inflammation induite par le formol et demono arthrite induite par la carraghénane/kaolinchez les rats.Par la suite, les bio-marqueurs du stress oxydatif ont été évalués en utilisant des dosages biochimiques standards.
Résultats: L’extrait n’a pas réduit la viabilité des cellules CHO-k1 et RAW 264.7.MEOg amontré despropriétés de stabilisation membranaires significatives (p<0,05).La MEOgà 100, 200 et 400 mg/kg réduit de façon dose-dépendante de lapatte d’œdème induitpar l’albumine de l’œufde 24,0, 32,2 et 37,8%, respectivement.De même, MEOg significativement (p <0,05) réduit
l’épaisseur de la patte par rapport au contrôle dans l’arthrite induite par le formol chez les rats.En outre,MEOg a dosedépendamment montré une activité anti-inflammatoire qui était évidente avec la diminution du gonflement des pattes et des genoux, et une diminution de la douleur inflammatoire dans lamono-arthriteinduite par la carraghénane /kaolinchez les rats.Le traitement a diminué les concentrations plasmatiques de TBARS et de NO et a augmenté les niveaux de GSH et de SOD.
Conclusion: Ces résultats soutiennent l’utilisation ethno-pharmacologique des feuilles d’ocimumgratissimumpour améliorer l’inflammation et la douleur de l’arthrite.
Mot-clés: Propriété cytotoxique, Antiarthritique, Ocimumgratissimum, carraghénane, kaolin
Correspondence: Prof. O.G. Ademowo, Department of Pharmacology and Therapeutics, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Ibadan, Nigeria. E-mail: ademowo g@yahoo.com
Reference
Mobasheri A. Intersection of Inflammation and Herbal Medicine in the Treatment of Osteoarthritis. Curr Rheum Reports.2012;14: 604-616.
Akinpelu AO, Maduagwu SM, Odole AC and Alonge TO. Prevalence and Pattern of knee osteoarthritis in a North Eastern Nigerian rural community. East Afri. Orthopaedic J. 2010; 5: 48-54.
Adebusoye LA, Ogunbode AM and Alonge TO. Magnitude of knee osteoarthritis and associated risk factors among adult patients presenting in a family practice clinic in Nigeria. J Med Trop. 2013; 15:144-150.
Breedveld FC. Current and Future management approaches for rheumatoid arthritis. Arthritis Res. 2002; 4: 16-21.
Recio MC, Andújar I and Ríos JL. Anti-Inflammatory Agents from Plants/ : Progress and Potential. Curr Medicinal Chem. 2012; 19: 2088-2103.
Curtis JR, Chastek B, Becker L, et al. Cost and effectiveness of biologics for rheumatoid arthritis in a commercially insured population. JManag Care Spec Pharm.2015; 21: 318-329.
Orwa C, Mutua A, Kindt R, Jamnadass R and Anthony S. Agroforestree Database: A tree reference and selection guide version 4.0. World Agroforestry Centre, Kenya. http://www.feedipedia.org/node/1650, 2009.
Aiyeloja AA and Bello OA. Ethnobotanical potential of Common herbs in Nigeria: A case study of Enugu State. Edu Res and Rev. 2006; 1: 16-22.
Iwu MM. Handbook of African Medicinal Plants. CRC Press, Boca Raton, Florida. 1993.
Ehiagbonare JE. Macropropagation of Ocimumgratissimum L: A multipurpose medicinal plant in Nigeria. Afri J Biotech.2007; 6: 013-014.
Ajayi AM, Tanayen JK, Ezeonwumelu JOC, et al. Anti-inflammatory, Anti-nociceptive and total polyphenolic content of hydroethanolic extract of Ocimum gratissimum L. Leaves. Afr J Med MedSci, 2014; 43s: 215-224.
NIH. Guide for the Care and use of laboratory animals. National Academic Press. 1996.
Zimmermann M. Ethical Guidelines for Investigations of Experimental Pain in Conscious Animals. Pain, 1983; 16:109-110.
Nakayima GR, Caton MC, Nova MP and Parandoosh Z. Assessment of the Alamar Blue assay for cellular growth and viability in vitro. J Immunol Methods. 1997; 204: 205-208.
Brown JH, Mackey HK and Rigglio DA. A Novel in vitro Assay for Anti-Inflammatory Agents Based on Stabilization of Erythrocytes. Exptal Biol Med. 1967; 125: 837.
Anosike CA, Obidoa O and Ezeanyika LU. Membrane stabilization as a mechanism of the anti-inflammatory activity of methanol extract of garden egg (Solanum aethiopicum). DARU J Pharmaceutical Sci. 2012; 20:76.
Akah PA and Nwambie IA. Evaluation of Nigerian traditional medicinal plants used for rheumatoid disorders. J Ethnopharamacol. 1994; 42: 179-182.
Sluka KA and Westlund KN. Behavioral and immunohistochemical changes in an experimental arthritis model in rats. Pain.1993; 55: 367-377.
Larsen JJ and Arnt J. Reduction in locomotor activity of arthritic rats as parameter for chronic pain: effect of morphine, acetylsalicylic acid and citalopram. Acta Pharmacol Toxicol. 1985; 57: 345-351.
Sin YM, Pook SH, Tan TM, et al. Changes in gluthathione and its associated enzymes during carrageenan-induced acute inflammation in mice. Comp Biochem Physiol. 1997; 116: 191-195.
Nagababu E, Rifkind JM, Sesikeran B and Lakshmaiah N. Assessment of antioxidant activities of eugenol by in vitro and in vivo methods. Methods in MolBiol (Clifton, N.J.) 2010; 610: 165-180.
Misra HP and Fridovich I. The Role of Superoxide Anion in the Autooxidation of epinephrine and a simple assay for Superoxide Dismutase. J Biol Chem.1972; 247: 3170-3175.
Gornall AG, Bradwill CJ and David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem.1949; 77:167-182.
Ahmed AS, McGaw LJ and Eloff JN. Evaluation of pharmacological activities, cytotoxicity and phenolic composition of four Maytenus species used in Southern African traditional medicine to treat intestinal infections and diarrhoeal diseases. BMC Complem and Alter Med. 2013; 13: 100.
Kpadonou Kpoviessi BGH, Kpoviessi SDS, Yayi Ladekan E, et al. In vitro antitrypanosomal and antiplasmodial activities of crude extracts and essential oils of Ocimum gratissimum Linn from Benin and influence of vegetative stage. J Ethnopharmacol. 2014;155: 1417-1423.
Stromstedt AA, Felth J and Bohlin. Bioassays in Natural Product Research – Strategies and Method in the search for anti-inflammatory and antimicrobial activity. Phytochemical analy. 2014; 25: 13-28.
Ignarro LJ. Effects of anti-inflammatory drugs on the stability of rat liver lysosomes in vitro. Biochem Pharmacol.1971; 20: 2847-2860.
Adzu B, Amos S, Dzarma S, Muazzam I and Gamaniel KS. Pharmacological evidence favouring the folkloric use of Diospyros mespiliformis Hochst in the relief of pain and fever. J. Ethnopharmacol.2001; 82: 191-195.
Greenwald RA. Animal models for evaluation of arthritic drugs. Method Find Clinical Pharmacol 1991; 13:75-83.
Akindele AJ and Adeyemi OO. Anti-inflammatory activity of the aqueous leaf extract of Byrsocarpus coccineus. Fitoterapia, 2007; 78: 25-28.
Bianchi M, Martucci C, Biella G, Ferrario P and Sacerdote P. Increased substance P and tumor necrosis factor-alpha level in the paws following formalin injection in rat tail. Brain Res. 2004; 1019:255-258.
Kim HD, Cho HR, Moon SB, et al. Effect of exopolymers from Aureobasidum pullulans on formalin-induced chronic paw inflammation in mice. J Microbiol Biotechnol. 2006; 16:1954-1960.
Kim BH, Kim M, Yin CH, et al. Inhibition of the signalling kinase JAK3 alleviates inflammation in monoarthritic rats. Br. J Pharmacol, 2011; 164: 106-118.
DiRosa M, Giroud JP and Willoughby DA. Studies of the acute inflammatory response induced in Rats in different sites by carrageenan and turpentine. J Pathol. 1971; 104: 15-29.
Salvemini DZQ, Wang PS, Wyatt DM, et al. Nitric oxide: a key mediator in the early and late phase of carrageenan-induced rat paws inflammation. Bri J Pharmacol.1996; 118: 829-838.
Egan CG, Lockhart JC, Ferrell WR, Day SM and Mclean JS. Pathophysiological basis of acute inflammatory hyperemia in the rat knee: roles of cyclooxygenase 1- and -2. J Physiol. 2002; 539: 579-587.
Ren K and Dubner R. Inflammatory Models of Pain and Hyperalgesia. ILAR J.1999; 40: 111-118.
Ogunnaike BF, Okutachi IR, Anucha ES, et al. Comparative anti-inflammatory activities of Jatropha curcas, Ocimum gratissimum and Solanum scabrum leaves. J Nat Prod Plant Resour. 2013; 3: 59-66.
Madhu KD and Harindran J. Anti-arthritic potential of Ocimum gratissimum L. In collagen induced arthritic Sprague-Dawley rats. Biomed Aging Pathol.2014; 4:191–196.
Zhang L, Zhang X and Westlund KN. Restoration of spontaneous exploratory behaviours with an intrathecal NMDA receptor antagonist or a PKC inhibitor in rats with acute pancreatitis. Pharmacol, Biochem, Behav. 2004; 77: 145-153.
Labuda CJ and Fuchs PN. A comparison of chronic aspartame exposure to aspirin on inflammation, hyperalgesia and open field activity following carrageenan-induced monoarthritis. Life Sci. 2001; 69: 443-454.
Millecamps M, Jourdan D, Leger S, et al. Circadian pattern of spontaneous behaviour in monarthritic rats: a novel global approach to evaluation of chronic pain and treatment effectiveness. Arthritis Rheum. 2005; 52: 3470-3478.
Matson DJ, Broom DC, Baldassari J, Kehne J and Cortright DN. Inflammation induced reduction of spontaneous activity by adjuvant: a novel model to study the effect of analgesics in rats. J Pharm Exp Ther. 2006; 320: 194-201.
Omote K, Hazama K, Kawamata T, et al. Peripheral Nitric Oxide In Carrageenan–Induced Inflammation. Brain Res. 2001; 912:171-175.
Bihani GV, Rojatkar SR and Bodhanker SL. Anti-arthritic activity of methanol extract of Cyathocline purpurea (Whole Plant) in Freund’s complete adjuvant –induced arthritis in rats. Biomed Aging Pathol. 2014; 4:197-206.