We examined the effects of a chloroform extract of Hyptis albida (CHA) on inflammatory responses in mouse lipopolysaccharide (LPS) induced peritoneal macrophages. Our findings indicate that CHA inhibits LPS-induced production of tumor necrosis factor (TNF-α) and interleukin-6 (IL-6). During the process, levels of cyclooxygenase-2 (COX-2), nitric oxide synthase (iNOS), and nitric oxide (NO) increased in the mouse peritoneal macrophages; however, the extract suppressed them significantly. These results provide novel insights into the anti-inflammatory actions of CHA and support its potential use in the treatment of inflammatory diseases. 1. Introduction Inflammation is an immediate response to many injuries produced by pathogens, noxious stimuli such as chemicals, or physical injury. Inflammation involves the activation and recruitment of phagocytes (macrophages, neutrophils), NK cells, the complement system, and the secretion of cytokines such as IL-1β, IL-6, and TNF-α by activated cells that are essential for the host defense system. Inflammatory disorders are treated using conventional anti-inflammatory drugs such as steroidal anti-inflammatory drugs and nonsteroidal anti-inflammatory drugs (NSAIDs) [1]. However, their prolonged use may produce adverse effects [2]. Thus, it is important to develop new anti-inflammatory agents with fewer adverse effects. Natural products can be a source of active metabolites that can serve as an alternate approach to anti-inflammatory drugs [3]. The genus Hyptis consists of approximately 400 species distributed from Southern United States to Argentina [4]. Plants in this genus have great economical and ethnopharmacological importance [5]. They have been used in folk medicine for the treatment of various disorders such as gastrointestinal disorders, skin infections, nasal congestion, fever, cramps, inflammation, and pain [5–8]. The genus Hyptis has many species that are important in Mexican folk medicine. In particular, Hyptis albida is commonly used in remedies for the treatment of gastrointestinal disturbances, skin infections, rheumatism, cramps, and muscular pains [9, 10]. Three triterpene lactones and five flavonoids have been isolated from an acetone extract [11] and the anti-inflammatory activity of a chloroform extract was reported by Pérez et al. [12]. The present investigation was carried out to assess the anti-inflammatory activity of a chloroform extract using murine macrophages stimulated with LPS. 2. Materials and Methods 2.1. Plant Material Aerial parts of H. albida were collected in Guadalcazar, San Luis
References
[1]
R. Gautam and S. M. Jachak, “Recent developments in anti-inflammatory natural products,” Medicinal Research Reviews, vol. 29, no. 5, pp. 767–820, 2009.
[2]
M. C. Allison, A. G. Howatson, C. J. Torrance, F. D. Lee, and R. I. Russell, “Gastrointestinal damage associated with the use of nonsteroidal antiinflammatory drugs,” The New England Journal of Medicine, vol. 327, no. 11, pp. 749–754, 1992.
[3]
M. S. Butler, “Natural products to drugs: natural product derived compounds in clinical trials,” Natural Product Reports, vol. 22, no. 2, pp. 162–195, 2005.
[4]
R. M. Harley, “Revision of generic limits in Hyptis Jacq. (Labiatae) and its allies,” Botanical Journal of the Linnean Society, vol. 98, no. 2, pp. 87–95, 1988.
[5]
C. R. P. Franco, ？. R. Antoniolli, A. G. Guimar？es et al., “Bioassay-guided evaluation of antinociceptive properties and chemical variability of the essential oil of Hyptis fruticosa,” Phytotherapy Research, vol. 25, no. 11, pp. 1693–1699, 2011.
[6]
M. D. Bispo, R. H. V. Mour？o, E. M. Franzotti et al., “Antinociceptive and antiedematogenic effects of the aqueous extract of Hyptis pectinata leaves in experimental animals,” Journal of Ethnopharmacology, vol. 76, no. 1, pp. 81–86, 2001.
[7]
A. X. Bueno, A. T. S. Moreira, F. T. Silva, C. S. Estevam, and M. Marchioro, “Effects of the aqueous extract from Hyptis pectinata leaves on rodent central nervous system,” Revista Brasileira De Farmacognosia, vol. 16, no. 3, pp. 317–323, 2006.
[8]
C. R. P. Franco, P. B. Alves, D. M. Andrade et al., “Essential oil composition and variability in Hyptis fruticosa,” Brazilian Journal of Pharmacognosy, vol. 21, no. 1, pp. 24–32, 2011.
[9]
M. Martínez, Catálogo De Nombres Vulgares Y Científicos De Plantas Mexicanas, Fondo de Cultura Económica, México, 1979.
[10]
J. L. Díaz, Uso De Las Plantas Medicinales De México, Instituto Mexicano para el Estudio de las Plantas Medicinales A.C., México, 1976.
[11]
R. Pereda-Miranda and G. Delgado, “Triterpenoids and flavonoids from Hyptis albida,” Journal of Natural Products, vol. 53, no. 1, pp. 182–185, 1990.
[12]
S. Pérez, B. L. Hernández, M. A. Zavala, G. E. Morales, and N. Cárdenas, “Antiinflammatory activity of Hyptis albida,” Journal of Medicinal Plants Research, vol. 6, no. 43, pp. 5582–5585, 2012.
[13]
X. A. Domínguez, Métodos De Investigación Fitoquímica, Ed Limusa México, 1973.
[14]
A. Elmann, S. Mordechay, H. Erlank, A. Telerman, M. Rindner, and R. Ofir, “Anti-Neuroinflammatory effects of the extract of Achillea fragrantissima,” BMC Complementary and Alternative Medicine, vol. 11, article 98, 2011.
[15]
K. S. Ahn, E. J. Noh, H. L. Zhao, S. H. Jung, S. S. Kang, and Y. S. Kim, “Inhibition of inducible nitric oxide synthase and cyclooxygenase II by Platycodon Grandiflorum saponins via suppression of nuclear factor-κB activation in RAW 264.7 cells,” Life Sciences, vol. 76, no. 20, pp. 2315–2328, 2005.
[16]
A. Sánchez-González, D. Granados-Sánchez, and R. Simón-Nabor, “Medicinal uses of plants by the otomi towership nicolas flores, hidalgo México,” Revista Chapingo Series Horticulture, vol. 14, no. 3, pp. 271–279, 2008.
[17]
N. T. Dung, V. K. Bajpai, J. I. Yoon, and S. C. Kang, “Anti-inflammatory effects of essential oil isolated from the buds of Cleistocalyx operculatus (Roxb.) Merr and Perry,” Food and Chemical Toxicology, vol. 47, no. 2, pp. 449–453, 2009.
[18]
D. B. Reddy and P. Reddanna, “Chebulagic acid (CA) attenuates LPS-induced inflammation by suppressing NF-κB and MAPK activation in RAW 264.7 macrophages,” Biochemical and Biophysical Research Communications, vol. 381, no. 1, pp. 112–117, 2009.
[19]
P.-H. Park, H. S. Kim, X. Y. Jin et al., “KB-34, a newly synthesized chalcone derivative, inhibits lipopolysaccharide-stimulated nitric oxide production in RAW 264.7 macrophages via heme oxygenase-1 induction and blockade of activator protein-1,” European Journal of Pharmacology, vol. 606, no. 1–3, pp. 215–224, 2009.
[20]
S.-I. Kanno, A. Shouji, A. Tomizawa et al., “Inhibitory effect of naringin on lipopolysaccharide (LPS)-induced endotoxin shock in mice and nitric oxide production in RAW 264.7 macrophages,” Life Sciences, vol. 78, no. 7, pp. 673–681, 2006.
[21]
I. Posadas, M. C. Terencio, I. Guillén et al., “Co-regulation between cyclo-oxygenase-2 and inducible nitric oxide synthase expression in the time-course of murine inflammation,” Naunyn-Schmiedeberg's Archives of Pharmacology, vol. 361, no. 1, pp. 98–106, 2000.
[22]
S. Moncada, R. M. J. Palmer, and E. A. Higgs, “Nitric oxide: physiology, pathophysiology, and pharmacology,” Pharmacological Reviews, vol. 43, no. 2, pp. 109–142, 1991.
[23]
D. Salvemini, Z.-Q. Wang, P. S. Wyatt et al., “Nitric oxide: a key mediator in the early and late phase of carrageenan-induced rat paw inflammation,” British Journal of Pharmacology, vol. 118, no. 4, pp. 829–838, 1996.
[24]
K. L. Davis, E. Martin, I. V. Turko, and F. Murad, “Novel effects of nitric oxide,” Annual Review of Pharmacology and Toxicology, vol. 41, pp. 203–236, 2001.
[25]
A. R. Amin, P. Vyas, M. Attur et al., “The mode of action of aspirin-like drugs: effect on inducible nitric oxide synthase,” Proceedings of the National Academy of Sciences of the United States of America, vol. 92, no. 17, pp. 7926–7930, 1995.
[26]
D. C. Rockey, J. J. Chung, C. M. McKee, and P. W. Noble, “Stimulation of inducible nitric oxide synthase in rat liver by hyaluronan fragments,” Hepatology, vol. 27, no. 1, pp. 86–92, 1998.
[27]
Q. Li and I. M. Verma, “NF-κB regulation in the immune system,” Nature Reviews Immunology, vol. 2, no. 10, pp. 725–734, 2002.
[28]
S. F. Liu and A. B. Malik, “NF-κB activation as a pathological mechanism of septic shock and inflammation,” American Journal of Physiology, vol. 290, no. 4, pp. L622–L645, 2006.
[29]
W. L. Smith, R. Michael Garavito, and D. L. DeWitt, “Prostaglandin endoperoxide H syntheses (cyclooxygenases)-1 and -2,” The Journal of Biological Chemistry, vol. 271, no. 52, pp. 33157–33160, 1996.
[30]
M. K. O'Banion, V. D. Winn, and D. A. Young, “cDNA cloning and functional activity of a glucocorticoid-regulated inflammatory cyclooxygenase,” Proceedings of the National Academy of Sciences of the United States of America, vol. 89, no. 11, pp. 4888–4892, 1992.
[31]
H. R. Herschman, “Prostaglandin synthase,” Biochimica et Biophysica Acta, vol. 1299, no. 1, pp. 125–140, 1996.
[32]
J. Clària, “Cyclooxigenase-2 biology,” Current Pharmaceutical Desing, vol. 9, pp. 2177–2190, 2003.
[33]
S. Zanotti, A. Kumar, and A. Kumar, “Cytokine modulation in sepsis and septic shock,” Expert Opinion on Investigational Drugs, vol. 11, no. 8, pp. 1061–1075, 2002.
[34]
M. A. West, S. C. Seatter, J. Bellingham et al., “Mechanisms of reprogrammed macrophage endotoxin signal transduction after lipopolysaccharide pretreatment,” Surgery, vol. 118, no. 2, pp. 220–228, 1995.
[35]
D. R. Hodge, E. M. Hurt, and W. L. Farrar, “The role of IL-6 and STAT3 in inflammation and cancer,” European Journal of Cancer, vol. 41, no. 16, pp. 2502–2512, 2005.
[36]
S. Mukhopadhyay, J. R. Hoidal, and T. K. Mukherjee, “Role of TNFα in pulmonary pathophysiology,” Respiratory Research, vol. 7, article 125, 2006.
[37]
T. Geiser, K. Atabai, P.-H. Jarreau, L. B. Ware, J. Pugin, and M. A. Matthay, “Pulmonary edema fluid from patients with acute lung injury augments in vitro alveolar epithelial repair by an IL-1β-dependent mechanism,” American Journal of Respiratory and Critical Care Medicine, vol. 163, no. 6, pp. 1384–1388, 2001.
[38]
P. M. Ridker, C. H. Hennekens, J. E. Buring, and N. Rifai, “C-reactive protein and other markers of inflammation in the prediction of cardiovascular disease in women,” The New England Journal of Medicine, vol. 342, no. 12, pp. 836–843, 2000.
[39]
S. Verma, S.-H. Li, M. V. Badiwala et al., “Endothelin antagonism and interleukin-6 inhibition attenuate the proatherogenic effects of C-reactive protein,” Circulation, vol. 105, no. 16, pp. 1890–1896, 2002.
[40]
B. Tarighi, T. Kurum, M. Demir, and S. N. Azcan, “The effects of nebivolol on fibrinolytic parameters in mild and moderate hypertensive patients,” Canadian Journal of Cardiology, vol. 23, no. 8, pp. 651–655, 2007.
[41]
D. Q. Falca？o and F. S. Menezes, “Revis？o etnofarmacológica, farmacológica e química do gênero Hyptis,” Revista Brasileira De Farmacognosia, vol. 84, no. 3, pp. 69–74, 2003.
[42]
Y. M. Fan, L. Z. Xu, J. Gao et al., “Phytochemical and antiinflammatory studies on Terminalia catappa,” Fitoterapia, vol. 75, no. 3-4, pp. 253–260, 2004.
[43]
N. Suh, T. Honda, H. J. Finlay et al., “Novel triterpenoids suppress inducible nitric oxide synthase (iNOS) and inducible cyclooxygenase (COX-2) in mouse macrophages,” Cancer Research, vol. 58, no. 4, pp. 717–723, 1998.
[44]
S. Y. Ryu, M.-H. Oak, S.-K. Yoon et al., “Anti-allergic and anti-inflammatory triterpenes from the herb of Prunella vulgaris,” Planta Medica, vol. 66, no. 4, pp. 358–360, 2000.
[45]
S. Shishodia, S. Majumdar, S. Banerjee, and B. B. Aggarwal, “Ursolic acid inhibits nuclear factor-κB activation induced by carcinogenic agents through suppression of IκBα kinase and p65 phosphorylation: correlation with down-regulation of cyclooxygenase 2, matrix metalloproteinase 9, and cyclin D1,” Cancer Research, vol. 63, no. 15, pp. 4375–4383, 2003.
[46]
T. Ishikawa, R. D. S. Donatini, I. E. C. Diaz, M. Yoshida, E. M. Bacchi, and E. T. M. Kato, “Evaluation of gastroprotective activity of Plinia edulis (Vell.) Sobral (Myrtaceae) leaves in rats,” Journal of Ethnopharmacology, vol. 118, no. 3, pp. 527–529, 2008.
[47]
S. J. Lee, R. H. Son, H. W. Chang, S. S. Kang, and H. P. Kim, “Inhibition of arachidonate release from rat peritoneal macrophage by biflavonoids,” Archives of Pharmacal Research, vol. 20, no. 6, pp. 533–538, 1997.
[48]
W. J. Kwak, T. C. Moon, C. X. Lin et al., “Papyriflavonol a from Broussonetia papyrifera inhibits the passive cutaneous anaphylaxis reaction and has a secretory phospholipase A2-inhibitory activity,” Biological and Pharmaceutical Bulletin, vol. 26, no. 3, pp. 299–302, 2003.
[49]
D. S. Jang, M. Cuendet, M. E. Hawthorne et al., “Prenylated flavonoids of the leaves of Macaranga conifera with inhibitory activity against cyclooxygenase-2,” Phytochemistry, vol. 61, no. 7, pp. 867–872, 2002.
[50]
R. Landolfi, R. L. Mower, and M. Steiner, “Modification of platelet function and arachidonic acid metabolism by bioflavonoids. Structure-activity relations,” Biochemical Pharmacology, vol. 33, no. 9, pp. 1525–1530, 1984.
[51]
T. Horie, M. Tsukayama, H. Kourai et al., “Syntheses of 5,6,7- and 5,7,8-trioxygenated 3',4'-dihydroxyflavones having alkoxy groups and their inhibitory activities against arachidonate 5-lipoxygenase,” Journal of Medicinal Chemistry, vol. 29, no. 11, pp. 2256–2262, 1986.
[52]
G. R. Reddy, N. Ueda, T. Hada et al., “A prenylflavone, artonin E, as arachidonate 5-lipoxygenase inhibitor,” Biochemical Pharmacology, vol. 41, no. 1, pp. 115–118, 1991.
[53]
M. J. Laughton, P. J. Evans, M. A. Moroney, J. R. S. Hoult, and B. Halliwell, “Inhibition of mammalian 5-lipoxygenase and cyclo-oxygenase by flavonoids and phenolic dietary additives. Relationship to antioxidant activity and to iron ion-reducing ability,” Biochemical Pharmacology, vol. 42, no. 9, pp. 1673–1681, 1991.
[54]
W. Krol, Z. P. Czuba, M. D. Threadgill, B. D. M. Cunningham, and G. Pietsz, “Inhibition of nitric oxide ( ) production in murine macrophages by flavones,” Biochemical Pharmacology, vol. 50, no. 7, pp. 1031–1035, 1995.
[55]
H. Sadowska-Krowicka, E. E. Mannick, P. D. Oliver et al., “Genistein and gut inflammation: role of nitric oxide,” Proceedings of the Society for Experimental Biology and Medicine, vol. 217, no. 3, pp. 351–357, 1998.
[56]
Y. Geng, B. Zhang, and M. Lotz, “Protein tyrosine kinase activation is required for lipopolysaccharide induction of cytokines in human blood monocytes,” Journal of Immunology, vol. 151, no. 12, pp. 6692–6700, 1993.
[57]
J. Y. Cho, P. S. Kim, J. Park et al., “Inhibitor of tumor necrosis factor-α production in lipopolysaccharide-stimulated RAW264.7 cells from Amorpha fruticosa,” Journal of Ethnopharmacology, vol. 70, no. 2, pp. 127–133, 2000.
