Flavonoid-Rich Fraction of Ocimum gratissimum Attenuates Lipopolysaccharide-Induced Sickness Behavior, Inflammatory and Oxidative Stress in Mice

Abstract

Purpose Ocimum gratissimum L. leaves has been traditionally used for management of febrile illnesses and symptoms typified of sickness behavior. In this work we investigated the modulatory effect of flavonoid-rich fraction of O. gratissimum leaves (EAFOg) on sickness behavior, inflammatory and oxidative stress responses in LPS-challenged mice.

Method O. gratissimum leaf was first extracted with n-hexane, chloroform and methanol, and EAFOg was obtained by ethylacetate partitioning of a sequentially resultant methanol extract. The effect of EAFOg (25–100 mg/kg) on acute LPS-induced neurobehavioral impairment in an open field test (OFT) and depressive-like behavior in forced swimming test (FST) was investigated. Serum nitrite and TNF-α, as well as myeloperoxidase (MPO), malondialdehyde (MDA), and reduced glutathione (GSH) levels were determined in liver and brain tissues.

Result EAFOg prevented the reduction in locomotor and rearing activity in OFT and the increase in immobility time in FST. The fraction significantly attenuated the elevation of serum TNF- α and nitrite levels. EAFOg reversed LPS-induced increase in MDA, MPO, and nitrite levels and attenuated GSH depletion in liver and brain tissues of mice.

Conclusion Flavonoid-rich fraction of O. gratissimum leaf demonstrated significant modulation of LPS-induced sickness behavior, inflammatory and oxidative stress response in mice. This suggests an important therapeutic strategy in slowing down LPS-mediated hepatic and neuronal disease processes.

Key words

Ocimum - sickness behaviour - lipopolysaccharides - flavonoids

• References

• 1 Dantzer R. Cytokine-induced sickness behavior: Mechanisms and implications. Ann N Y Acad Sci 2001; 933: 222-234
• 2 Viljoen M, Panzer A. Sickness behavior: Causes and Effects. SA Fam Pract 2003; 45: 15-18
• 3 Moon ML, McNeil LK, Freund GG. Macrophages make me sick: How macrophage activation states influence sickness behavior. Psychoneuroendocrinol 2011; 36: 1431-1440
• 4 Dantzer R, O'Connor JC, Freund GG. et al. From inflammation to sickness and depression: When the immune system subjugates the brain. Nat Rev Neurosci 2008; 9: 46e56
• 5 McCusker RH, Kelley KW. Immune-neural connections: how the immune system's response to infectious agents influences behavior. J Exp Biol. 2013; 216: 84e98
• 6 Araki R, Hiraki Y, Nishida S. et al. Gomisin N ameliorates lipopolysaccharide-induced depressive-like behaviors by attenuating inflammation in the hypothalamic paraventricular nucleus and central nucleus of the amygdala in mice. J Pharmacological Sci 2016; 132: 138-144
• 7 Becskei C, Riediger T, Hernádfalvy N. et al. Inhibitory effects of lipopolysaccharide on hypothalamic nuclei implicated in the control of food intake. Brain Behav Immun 2008; 22: 56-64
• 8 Biswas SK. Does the interdependence between oxidative stress and inflammation explain the antioxidant paradox?. Oxidative Medicine and Cellular Longevity 2016 Article ID 5698931 9
• 9 Flohé L, Brigelius-Flohé R, Saliou C. et al. “Redox regulation of NF-κB activation,”. Free Rad Biol Med 1997; 22: 1115-1126
• 10 Prabhu KS, Loboi R, Shirwaikar AA. et al. Ocimum gratissimum: A review of its chemical, pharmacological and ethnomedicinal properties. The Open Complement Med J 2009; 1: 1-15
• 11 El Said F, Sofowora EA, Malcolm SA. et al. An investigation into the efficacy of Ocimum gratissimum as used in Nigerian native medicine. Planta Medica 1969; 17: 195 -200
• 12 Iwu MM. Handbook of African Medicinal Plants. CRC Press; Boca Raton, Florida: 1993
• 13 Ajibesin KK, Ekpo BA, Bala DN. et al. Ethnobotanical survey of Akwa Ibom State of Nigeria. J Ethnopharmacol 2008; 115: 387-408
• 14 Ajaiyeoba EO, Oladepo O, Fawole OI. et al. Cultural categorization of febrile illnesses in correlation with herbal remedies used for treatment in Southwestern Nigeria. J Ethnopharmacol 2003; 85: 179-185
• 15 Ebong OO, Ajaiyeoba EO, Ononiwu IM. et al. Contribution of indigenous health caregivers to the herbal management of febrile illnesses in Rivers state, South-south, Nigeria. W Afri J Pharmacol Drug Res 2005; 21: 48-54
• 16 Paton A. A synopsis of Ocimum L. (Labiatae) in Africa. Kew Bulletin 1992; 473: 403-435
• 17 Grayer RJ, Kite GC, Abou-Zaid M. et al. The application of atmospheric pressure chemical ionisation liquid chromatography-Mass Spectrometry In the chemotaxonomic study of flavonoids: Characterisation of flavonoids from Ocimum gratissimum L. Var gratissimum. Phytochem Analy 2002; 11: 257-267
• 18 Ola SS, Catia G, Marzia I. et al. HPLC/DAD/MS characterisation and analysis of flavonoids and cynnamoil derivatives in four Nigerian green-leafy vegetables. Food Chem 2009; 115: 1568-1574
• 19 Gupta A, Vij G, Sharma S. et al. Curcumin, a polyphenolic antioxidant, attenuates chronic fatigue syndrome in murine water immersion stress model. Immunobiol 2009; 214: 33-39
• 20 Ajayi AM, Ologe MO, Ben-Azu B. et al. Ocimum gratissimum Linn. Leaf extract inhibits free radical generation and suppressed inflammation in carrageenan-induced inflammation models in rats. J Basic Clin Physiol Pharmacol 2017; 28: 531-541
• 21 Granato D, Karnopp AR, Van Ruth SM. Characterization and comparison of phenolic composition, antioxidant capacity and instrumental taste profile of Juices from different botanical origins. J Sci, Food and Agric 2015; 95: 1997-2006
• 22 Heinrich M, Gibbons S. Ethnopharmacology in drug discovery: an analysis of its role and potential contribution. J Pharm and Pharmacol 2001; 53: 425 -432
• 23 Ajayi AM, Martins DTO, Balogun SO. et al. Ocimum gratissimum L. leaf flavonoid-rich fraction suppress LPS-induced inflammatory response in RAW 264.7 macrophages and peritonitis in mice. J Ethnopharmacol 2017; 204: 169-178
• 24 NIH. Guide for the Care and use of laboratory animals. National Academic Press; 1996
• 25 Custodio CS, Mello BSF, Cordeiro RC. et al. Time course of the effects of lipopolysaccharide on prepulse inhibition and brain nitrite content in mice. Euro J Pharmacol 2013; 713: 31-38
• 26 Ribeiro DE, Maiolini VM, Soncini R. et al. Inhibition of nitric oxide synthase accentuate endotoxin-induced behaviour in mice. Pharmacol, Bioch Beh 2008; 103: 535-540
• 27 de Paiva VN, Lima SN, Fernandes MM. et al. Prostaglandins mediate depressive-like behaviour induced by endotoxin in mice. Behav Brain Res 2010; 215: 146-151
• 28 Bailey KR, Crawley JN. Methods of Behavior Analysis in Neuroscience. 2nd Florida: CRC Press; 2009
• 29 Soncini R, Souzaa DF, Neves AP. et al. Dipyrone attenuates sickness response to lipopolysaccharide in mice. Neurosci Lett 2012; 516: 114-118
• 30 Green LC, Wagner DA, Godowsky J. et al. Analysis of nitrate, nitrite and [15 N] nitrate in biological fluids. Anal Biochem 1982; 126: 131-138
• 31 Sin YM, Pook SH, Tan TM. et al. Changes in glutathione and its associated enzymes during carrageenan-induced acute inflammation in mice. Comp Biochem Physiol 1997; 116: 191-195
• 32 Nagababu E, Rifkind JM, Sesikeran B. et al. Assessment of antioxidant activities of eugenol by in vitro and in vivo methods. Methods in Mol Biol (Clifton, N.J.) 2010; 610: 165-180
• 33 Gornall AG, Bradwill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem 1949; 77: 167-182
• 34 Bradley PP, Christensen RD, Rothestein G. Cellular and extracellular myeloperoxidase in pyogenic inflammation. Blood 1982; 60: 618-622
• 35 Godbout JP, Chen J, Abraham J. et al. Exaggerated neuroinflammation and sickness behavior in aged mice following activation of the peripheral innate immune system. FASEB J. 2005; 19: 1329-1331
• 36 Ohgi Y, Futamura T, Kikuchi T. et al. Effects of antidepressants on alternations in serum cytokines and depressive-like behavior in mice after lipopolysaccharide administration. Pharmacol. Biochem. Behav 2013; 103: 853-859
• 37 Konsman JP, Parnet P, Dantzer R. Cytokine-induced sickness behavior: mechanisms and implications. Trends Neurosci 2002; 25: 154-159
• 38 Yamagata K, Matsumura K, Inoue W. et al. Co-expression of microsomal-type prostaglandin E synthase with cyclooxygenase-2 in brain endothelial cells of rats during endotoxin-induced fever. J Neurosci 2001; 21: 2669-2677
• 39 Bluthe RM, Laye S, Michaud B. et al. Role of interleukin-1beta and tumour necrosis factor-alpha in lipopolysaccharide induced sickness behavior: a study with interleukin-1 type I receptor deficient mice. Eur J Neurosci 2000; 12: 4447-4456
• 40 Engeland CG, Nielsen DV, Kavaliers M. et al. Locomotor activity changes following lipopolysaccharide treatment in mice: A multivariate assessment of behavioral tolerance. Physiol Behav. 2001; 72: 481-491
• 41 Krishna S, Dodd CA, Filipov NM. Behavioral and monoamine perturbations in adult male mice with chronic inflammation induced by repeated peripheral lipopolysaccharide administration. Behav Brain Res. 2016; 302: 279-290
• 42 Swiergiel AH, Dunn AJ. Effects of Interleukin-1β and Lipopolysaccharide on Behavior of Mice in the Elevated Plus-Maze and Open Field Tests. Pharmacol Biochem Beh 2007; 86: 651-659
• 43 Dunn AJ, Swiergiel AH. Effects of interleukin-1 and endotoxin in the forced swim and tail suspension tests in mice. Pharmacol Biochem Behav 2005; 81: 688-693
• 44 Harrison NA, Brydon L, Walker C. et al. Inflammation causes mood changes through alterations in subgenual cingulate activity and mesolimbic connectivity. Biol Psychiatry 2009; 66: 407-414
• 45 Eisenberger N, Inagaki T, Mashal N. et al. Inflammation and social experience: an inflammatory challenge induces feelings of social disconnection in addition to depressed mood. Brain Behav Immun 2010; 24: 558-563
• 46 Harada S, Imaki T, Chikada N. Distinct distribution and time-course changes in neuronal nitric oxide synthase and inducible NOS in the paraventricular nucleus following lipopolysaccharide injection. Brain Res 1999; 821: 322-332
• 47 Connor TJ, O'Sullivan J, Nolan Y. et al. Inhibition of constitutive nitric oxide production increases the severity of lipopolysaccharide-induced sickness behaviour: A role for TNF-alpha. Neuroimmunomodulation 2002; 10: 367-378
• 48 Reino DC, Pisarenko V, Palange D. et al. Trauma hemorrhagic shock-induced lung injury involves a gut-lymph-induced TLR4 pathway in mice. PLoS One 2011; 6: e14829 10.1371/journal.pone.0014829
• 49 Klebanoff SJ. Myeloperoxidase: Friend and foe. J Leuk Biol 2005; 77: 598-625
• 50 van Antwerpen P, Dufrasne F, Lequeux M. et al. Inhibition of the myeloperoxidase chlorinating activity by non-steroidal anti-inflammatory drugs: Flufenamic acid and its 5-chloro-derivative directly interact with a recombinant human myeloperoxidase to inhibit the synthesis of hypochlorous acid. Eur J Pharmacol 2007; 570: 235-243
• 51 Johri A, Beal FM. Antioxidants in Huntington's disease. Biochim Biophys Acta 2012; 1822: 664-674
• 52 Valko M, Leibfritz D, Moncol J. et al. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007; 39: 44-84
• 53 Pandya CD, Howell KR, Pillai A. Antioxidants as potential therapeutics for neuropsychiatric disorders. Prog. Neuro-Psychopharmacol Biol Psychiatry 2013; 46: 214-223
• 54 Peristeris P, Clark BD, Gatti S. et al. N-acetylcysteine and glutathione as inhibitors of tumor necrosis factor production. Cell Immunol 1992; 140: 390-399
• 55 Villa P, Saccani A, Sica A. et al. Glutathione Protects Mice from Lethal Sepsis by Limiting Inflammation and Potentiating Host Defense. J Infect Dis 2002; 185: 1115-1120
• 56 Abdel-Salam OME, Youness ER, Mohammed NA. et al Neuroprotective and hepatoprotective effects of micronized purified flavonoid fraction (Daflon®) in Lipopolysaccharide-treated rats. Drug Discov & Therap 2012; 6: 306-314