Abstract
Rotula aquatica belongs to the family Boraginaceae, and is reported to contain baunerol, steroids and alkaloids. In Ayurveda, R. aquatica has been used for the treatment of various diseases such as diabetes, treatment of piles, venereal disease, and cancer. The current study aims to investigate the anti-inflammatory effect of methanolic extract of R. aquatica (MERA) in RAW 264.7 cells. The cytotoxicity of MERA was analyzed by MTT assay. The total cyclooxygenase (COX) activity, 5-lipoxygenase (5-LOX) activity, myeloperoxidase activity, inducible nitric oxide synthase activity, nitrate level and reactive oxygen species production were studied in LPS-stimulated RAW 264.7 cells. The gene level expression of cyclooxygenase-2 (COX-2), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) were also evaluated in this study. The MERA did not show any cytotoxicity at different concentrations (6.25–100 µg/ml). MERA (100 μg/ml) inhibited total COX and 5-LOX activity at 50.53 and 62.03%, respectively, besides significantly (p < 0.05) diminished nitrate and ROS generation, when compared with LPS control. Moreover, MERA down-regulated the mRNA expressions of inflammatory marker genes like TNF-α, IL-6, and COX-2 against LPS stimulation. Our results demonstrate that MERA is able to attenuate inflammatory response, possibly via ROS and NO suppression, inhibiting the production of arachidonic acid metabolites and modulation of proinflammatory mediators and cytokines release.
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References
Axelrod B, Cheesbrough TM, Laakso S (1981) Lipoxygenase from soybeans. EC 1.13.11.12 Linoleate: oxygen oxidoreductase. Methods Enzymol 71:441–451. https://doi.org/10.1016/0076-6879(81)71055-3
Bistrian B (2007) Systemic response to inflammation. Nutr Rev 65:S170–S172
Bose S, Kim H (2013) Evaluation of in vitro anti-inflammatory activities and protective effect of fermented preparations of rhizoma atractylodis macrocephalae on intestinal barrier function against lipopolysaccharide insult. eCAM 2013:363076. https://doi.org/10.1155/2013/363076
Breckwoldt MO, Chen JW, Stangenberg L et al (2008) Tracking the inflammatory response in stroke in vivo by sensing the enzyme myeloperoxidase. Proc Natl Acad Sci USA 105:18584–18589. https://doi.org/10.1073/pnas.0803945105
Bryan NS, Grisham MB (2007) Methods to detect nitric oxide and its metabolites in biological samples. Free Radical Biol Med 43:645–657. https://doi.org/10.1016/j.freeradbiomed.2007.04.026
Christina AJM, Najumadeen NAH, Kumar SV et al (2008) Antilithiatic effect of Melia azedarach on ethylene glycol-induced nephrolithiasis in rats. Pharm Biol 44:480–485
Dillingh MR, van Poelgeest EP, Malone KE et al (2014) Characterization of inflammation and immune cell modulation induced by low-dose LPS administration to healthy volunteers. J Inflamm 11:28. https://doi.org/10.1186/s12950-014-0028-1
Epstein FH, Barnes PJ, Karin M (1997) Nuclear factor-κB—a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med 336:1066–1071. https://doi.org/10.1056/NEJM199704103361506
Esposito E, Cuzzocrea S (2009) TNF-alpha as a therapeutic target in inflammatory diseases, ischemia-reperfusion injury and trauma. Curr Med Chem 16:3152–3167
Fujiwara N, Kobayashi K (2005) Macrophages in inflammation. Curr Drug Targets Inflamm Allergy 4:281–286
García JJ, López-Pingarrón L, Almeida-Souza P et al (2014) Protective effects of melatonin in reducing oxidative stress and in preserving the fluidity of biological membranes: a review. J Pineal Res 56:225–237. https://doi.org/10.1111/jpi.12128
Girard-Lalancette K, Pichette A, Legault J (2009) Sensitive cell-based assay using DCFH oxidation for the determination of pro- and antioxidant properties of compounds and mixtures: analysis of fruit and vegetable juices. Food Chem 115:720–726. https://doi.org/10.1016/j.foodchem.2008.12.002
Heo S-J, Jang J, Ye B-R et al (2014) Chromene suppresses the activation of inflammatory mediators in lipopolysaccharide-stimulated RAW 264.7 cells. Food Chem Toxicol 67:169–175. https://doi.org/10.1016/j.fct.2014.02.023
Hinz B, Brune K (2002) Cyclooxygenase-2–10 years later. J Pharmacol Exp Therap 300:367–375
Kang Y-J, Mbonye UR, DeLong CJ et al (2007) Regulation of intracellular cyclooxygenase levels by gene transcription and protein degradation. Prog Lipid Res 46:108–125. https://doi.org/10.1016/j.plipres.2007.01.001
Klampfer L (2011) Cytokines, inflammation and colon cancer. Curr Cancer Drug Targets 11:451–464. https://doi.org/10.2174/156800911795538066
Kwang Seok A, Aggarwal BB (2005) Transcription factor NF-κB: a sensor for smoke and stress signals. In: Annals of the New York Academy of Sciences, pp 218–233
Lanata A, Aquatica R, Singh LS et al (2011) Comparative study of anthelmintic activity between aqueous extract of Areva lanata and Rotula aquatica lour. Asian J Pharmacy Life Sci ISSN 1:2231–4423
Lawrence T, Willoughby DA, Gilroy DW (2002) Anti-inflammatory lipid mediators and insights into the resolution of inflammation. Nat Rev Immunol 2:787–795. https://doi.org/10.1038/nri915
Leskovac A, Joksic G, Pasti I et al (2013) The antiradical, anti-inflammatory and anti-genotoxic potential of herbal preparation Chlamyfin. Maced J Chem Chem Eng 32:227–237. https://doi.org/10.20450/MJCCE.2013.192
Lin C-Y, Lee C-H, Chang Y-W et al (2014) Pheophytin a inhibits inflammation via suppression of LPS-induced nitric oxide synthase-2, prostaglandin E2, and interleukin-1β of macrophages. Int J Mol Sci 15:22819–22834. https://doi.org/10.3390/ijms151222819
Martel-Pelletier J, Lajeunesse D, Reboul P, Pelletier J-P (2003) Therapeutic role of dual inhibitors of 5-LOX and COX, selective and non-selective non-steroidal anti-inflammatory drugs. Ann Rheum Dis 62:501–509
Mogensen TH (2009) Pathogen recognition and inflammatory signaling in innate immune defenses. Clin Microbiol Rev 22:240–73, Table of Contents. https://doi.org/10.1128/CMR.00046-08
Molloy RG, Mannick JA, Rodrick ML (1993) Cytokines, sepsis and immunomodulation. Br J Surg 80:289–297
Nastasijević B, Lazarević-Pašti T, Dimitrijević-Branković S et al (2012) Inhibition of myeloperoxidase and antioxidative activity of Gentiana lutea extracts. J Pharm Biomed Anal 66:191–196. https://doi.org/10.1016/j.jpba.2012.03.052
Niu X, Xing W, Li W et al (2012) Isofraxidin exhibited anti-inflammatory effects in vivo and inhibited TNF-α production in LPS-induced mouse peritoneal macrophages in vitro via the MAPK pathway. Int Immunopharmacol 14:164–171. https://doi.org/10.1016/j.intimp.2012.06.022
Patil S, Narayanan S, Eibl G, Jolly CI (2004) Evaluation of antimitotic activity of Rotula aquatica (Lour): a traditional herb used in treatment of cancer. Indian J Exp Biol 42:893–899
Peskar BM (2001) Role of cyclooxygenase isoforms in gastric mucosal defence. J Physiol Paris 95:3–9
Pulli B, Ali M, Forghani R et al (2013) Measuring myeloperoxidase activity in biological samples. PLoS ONE 8:e67976. https://doi.org/10.1371/journal.pone.0067976
Rathore B, Ali Mahdi A, Nath Paul B et al (2007) Indian herbal medicines: possible potent therapeutic agents for rheumatoid arthritis. J Clin Biochem Nutr 41:12–17. https://doi.org/10.3164/jcbn.2007002
Ricciotti E, Fitzgerald GA (2011) Prostaglandins and inflammation. Arterioscler Thromb Vasc Biol 31:986–1000. https://doi.org/10.1161/ATVBAHA.110.207449
Saklatvala J, Dean J, Clark A (2003) Control of the expression of inflammatory response genes. In: Biochemical Society symposium, pp 95–106
Salter M, Duffy C, Garthwaite J, Strijbos PJ (1996) Ex vivo measurement of brain tissue nitrite and nitrate accurately reflects nitric oxide synthase activity in vivo. J Neurochem 66:1683–1690
Savva A, Roger T (2013) Targeting toll-like receptors: promising therapeutic strategies for the management of sepsis-associated pathology and infectious diseases. Front Immunol 4:387. https://doi.org/10.3389/fimmu.2013.00387
Shimizu T, Kondo K, Hayaishi O (1981) Role of prostaglandin endoperoxides in the serum thiobarbituric acid reaction. Arch Biochem Biophys 206:271–276. https://doi.org/10.1016/0003-9861(81)90091-6
Shirumalla RK, Naruganahalli KS, Dastidar SG et al (2006) RBx 7796: a novel inhibitor of 5-lipoxygenase. Inflamm Res 55:517–527. https://doi.org/10.1007/s00011-006-6032-z
Shivkar YM, Kumar VL (2003) Anthelmintic activity of latex of Calotropis procera. Pharm Biol 41:263–265. https://doi.org/10.1076/phbi.41.4.263.15666
Sollman T (1918) Anthelmintics: their efficiency as tested on earthworms. J Pharmacol Exp Ther 12:129–170
Süleyman H, Demircan B, Karagöz Y (2007) Anti-inflammatory and side effects of cyclooxygenase inhibitors. Pharmacol Rep 59:247–258
Suzuki K, Ota H, Sasagawa S et al (1983) Assay method for myeloperoxidase in human polymorphonuclear leukocytes. Anal Biochem 132:345–352
Takeda K, Akira S (2005) Toll-like receptors in innate immunity. Int Immunol 17:1–14. https://doi.org/10.1093/intimm/dxh186
Talarico LB, Zibetti RGM, Faria PCS et al (2004) Anti-herpes simplex virus activity of sulfated galactans from the red seaweeds Gymnogongrus griffithsiae and Cryptonemia crenulata. Int J Biol Macromol 34:63–71. https://doi.org/10.1016/j.ijbiomac.2004.03.002
Tanaka T, Narazaki M, Kishimoto T (2012) Therapeutic targeting of the interleukin-6 receptor. Annu Rev Pharmacol Toxicol 52:199–219. https://doi.org/10.1146/annurev-pharmtox-010611-134715
Umesh Kr G, Ajm C (2011) Effect of Rotula aquatica Lour. on ethylene-glycol induced urolithiasis in rats. Int J Drug Dev Res 3:273–280
Vieira TO, Seifriz I, Charão CCT et al (2011) Antioxidant effects of crude extracts from Baccharis species: inhibition of myeloperoxidase activity, protection against lipid peroxidation, and action as oxidative species scavenger. Rev Bras Farm 21:601–607. https://doi.org/10.1590/S0102-695X2011005000091
West MA, Seatter SC, Bellingham J, Clair L (1995) Mechanisms of reprogrammed macrophage endotoxin signal transduction after lipopolysaccharide pretreatment. Surgery 118:220–228
Zhao F, Wang L, Liu K (2009) In vitro anti-inflammatory effects of arctigenin, a lignan from Arctium lappa L., through inhibition on iNOS pathway. J Ethnopharmacol 122:457–462. https://doi.org/10.1016/j.jep.2009.01.038
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The authors are grateful to the School of Biosciences, Mahatma Gandhi University and Biogenix Research Center, Poojapura, Thiruvananthapuram, India for providing the excellent research support.
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Vysakh, A., Gopika, P., Jayesh, K. et al. Rotula aquatica Lour attenuates secretion of proinflammatory mediators and cytokines in lipopolysaccharide-induced inflammatory responses in murine RAW 264.7 macrophages. Inflammopharmacol 26, 29–38 (2018). https://doi.org/10.1007/s10787-017-0420-6
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DOI: https://doi.org/10.1007/s10787-017-0420-6