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Protective effect of 5-hydroxy-3′,4′,7-trimethoxyflavone against inflammation induced by lipopolysaccharide in RAW 264.7 macrophage: in vitro study and in silico validation

Abstract

The herb Lippia nodiflora L. (Verbenaceae) has been documented to exhibit anti-inflammatory, antipyretic, antitussive, antidiabetic, anticancer, and antimelanogenesis properties. In the present study, we aimed at evaluating the anti-inflammatory activity of 5-hydroxy-3′,4′,7-trimethoxyflavone, a flavonoid from L. nodiflora, using lipopolysaccharide induced inflammation in RAW 264.7 macrophages. 5-hydroxy-3′,4′,7-trimethoxyflavone significantly inhibited nitric oxide production and demonstrated slight reduction in prostaglandin-E2 level at tested concentrations. The production of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-1β, were obviously reduced by 5-hydroxy-3′,4′,7-trimethoxyflavone in a concentration-dependent manner. Moreover, 5-hydroxy-3′,4′,7-trimethoxyflavone significantly induced reduction in the mRNA expressions of inducible nitric oxide synthase and cyclooxygenase-2, representing that inhibition occurs at the transcriptional level. In addition, we performed molecular docking and molecular dynamic simulations to study the interaction of 5-hydroxy-3′,4′,7-trimethoxyflavone with inflammatory mediators such as inducible nitric oxide synthase and cyclooxygenase-2. Docking study showed its hydrogen bond interactions with key residues in the active site of inducible nitric oxide synthase and cyclooxygenase-2, enlightening its possible binding mode at the molecular level. The results of molecular dynamic simulations showed the stability of complexes and their interactions. Taken together, these findings envisage 5-hydroxy-3′,4′,7-trimethoxyflavone as a potential candidate molecule for the progress of therapeutic strategy against inflammation-related diseases.

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Acknowledgments

We gratefully acknowledge the financial assistance given by Department of Science and Technology (DST WOS-A), New Delhi, for granting fellowship to A. Sudha (Grant No. SR/WOS-A/LS-237/2010). The authors thank Dr. B. Kadalmani, Department of Animal Science, Bharathidasan University, Tiruchirapalli for providing help in in vitro studies.

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Correspondence to Pappu Srinivasan.

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Sudha, A., Jeyakanthan, J. & Srinivasan, P. Protective effect of 5-hydroxy-3′,4′,7-trimethoxyflavone against inflammation induced by lipopolysaccharide in RAW 264.7 macrophage: in vitro study and in silico validation. Med Chem Res 25, 1754–1767 (2016). https://doi.org/10.1007/s00044-016-1611-1

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Keywords

  • 5-hydroxy-3′,4′,7-trimethoxyflavone
  • Inflammation
  • Cytokines
  • COX-2
  • iNOS
  • Molecular docking