In Vivo and In Vitro Immunomodulatory Potential of Swertiamarin Isolated from Enicostema axillare (Lam.) A. Raynal That
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In Vivo and In Vitro Immunomodulatory Potential of Swertiamarin Isolated from Enicostema axillare (Lam.) A. Raynal That Acts as an Anti-inflammatory Agent S. Saravanan,1 P. Pandikumar,1 N. Prakash Babu,1 V. I. Hairul Islam,2,3 K. Thirugnanasambantham,3 M. Gabriel Paulraj,1 K. Balakrishna,1 and S. Ignacimuthu1,4
Abstract—Swertiamarin is a secoiridoid glycoside found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used as a depurative in the Indian system of traditional medicine. The present study evaluated the immunomodulatory activity of isolated swertiamarin. In vivo immunomodulatory activity of swertiamarin (2, 5, and 10 mg/kg b.w.) was evaluated in a model of sheep red blood cells (SRBC) by assessing its effect on organ weight, hemagglutinating antibody titer (HA), plaque-forming cells (PFC), quantitative hemolysis of SRBC, and delayed type hypersensitivity (DTH). In vitro immunomodulatory potential was studied on isolated splenocytes, neutrophils, and peritoneal macrophages. In silico immunomodulatory effects were evaluated by docking of swertiamarin on proinflammatory cytokines to confirm its potential. In in vivo studies, the animals treated with swertiamarin showed a significant (P≤ 0.05) increase in antibody titer, plaque-forming cells, and also in weight of the thymus and spleen. A decreased response to DTH reaction was recorded with the treatment of swertiamarin. In in vitro studies, treatment with swertiamarin modulated the messenger RNA (mRNA) and protein expression of IFN-γ, IL-10, and IL-4 significantly (P≤0.05) and also favored Th2-mediated response on concanavalin A (Con A)-induced splenocytes. The compound inhibited the release of free radicals significantly (P≤0.05) in phytohemagglutinin (PHA)-induced neutrophils and also ameliorated the mRNA and protein expression of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) in lipopolysaccharide (LPS)-induced macrophages. In in silico, the best docked pose of swertiamarin with the target proteins (TNF-α, IL-1β, and IL6) was confirmed that swertiamarin acted as an anti-inflammatory mediator. KEY WORDS: swertiamarin; immunomodulation; Th1/Th2-mediated response; molecular docking; antiinflammatory agent.
INTRODUCTION Immune cells are like double-edged swords that play a vital role in defense against pathogens; but in some conditions, they induce autoimmune diseases through undesirable immune reaction. These undesirable immune responses cause inflammation/tissue damage. Excessive 1
Division of Ethnopharmacology, Entomology Research Institute, Loyola College, Chennai, 600 034, Tamil Nadu, India 2 Division of Microbiology, Entomology Research Institute, Loyola College, Chennai, 600 034, Tamil Nadu, India 3 Pondicherry Centre for Biological Sciences, Pondicherry, 605 005, India 4 To whom correspondence should be addressed at Division of Ethnopharmacology, Entomology Research Institute, Loyola College, Chennai, 600 034, Tamil Nadu, India. E-mail: [email protected]
or aberrant inflammation proceeds to a chronic state, assoc i a t e d w i t h u n d
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