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Evaluation of the Anti-Inflammatory Activity of Kokilaksham kashayam on Lipopolysaccharide Stimulated RAW 264.7 Macrophages

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

The immune system involves a complex mechanism that operates in an ochestered manner protecting the host against the causative agents of injury. But in pathophysiological conditions, there occurs an unregulated production of pro-inflammatory cytokines- tumor necrosis factor (TNF-α) and interleukin (IL-1β) by cells like monocytes, macrophages and dendritic cells. Moreover, it is seen that several biochemical components such as lipid, protein, and nucleic acids also show variation in a diseased state. The present study aims to evaluate the effect of Kokilaksham Kashayam (KK) on the production of pro-inflammatory cytokines by lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage cell line. KK is an ayurvedic decoction traditionally used to treat diseases like rheumatoid arthritis, gout, jaundice, and anemia but the mechanism of its action in controlling inflammatory conditions has not been explored. The present methodology used involved the quantification of cytokines by ELISA method and gene expression studies by using qRTPCR followed by determining the effect of KK on the cellular changes caused due to LPS stimulation by Raman spectroscopic studies. The study revealed that KK could lower the increased levels of pro-inflammatory cytokines, which may be due to the downregulation of the genes for TNF-α and IL-1β. The results of Raman spectra further supported the results obtained for LPS stimulated RAW 264.7 cells pretreated with the fractions of KK. These findings provide scientific validation of the mechanism of action of KK, proving its anti-inflammatory potential. Further identification of the bioactive molecules of the decoction would help to develop newer therapeutic strategies for controlling chronic inflammatory conditions.

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Acknowledgements

This work was carried out with the support of the Kerala State Council for Science Technology and Environment (KSCSTE Order No.235/ WSD-BLP/KSCSTE/2016-17). The authors are grateful to DBT- MSUB- IPLSARE (BUILDER) programme (BT/PR4800/INF/22/152/2012 dated 22/03/2012), School of Biosciences, Mahatma Gandhi University, Kerala, for providing instrument facilities. The authors would like to thank Arya Vaidya Sala Kottakkal, Kerala, for their support.

Funding

Women Scientist division, Kerala State Council for Science Technology and Environment (KSCSTE Order No.235/ WSD-BLP/KSCSTE/2016–17), Thiruvanathapuram, Kerala.

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Authors and Affiliations

  1. Inflammation Research Lab, School of Biosciences, M.G.University, Kottayam, Kerala, 686560, India

    Teena Merlin, M. B. Bashi, S. J. Rahitha Devi & B. Prakash Kumar

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  1. Teena Merlin
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  2. M. B. Bashi
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  3. S. J. Rahitha Devi
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  4. B. Prakash Kumar
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Contributions

PKB and TM designed the research work. TM performed the experiments, analyzed the data. TM, BMB, and RDSJ interpreted the data and edited the manuscript. The final version of the manuscript was read and approved by all authors.

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Correspondence to B. Prakash Kumar.

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Significance statement

Ayurveda, the ancient traditional medical system comprises of herbal medicines widely recommended for the treatment of chronic inflammatory conditions. This work is designed to provide scientific validation of the mechanism of action of Kokilaksham kashayam, an ayurvedic herbal preparation proving its anti-inflammatory potential.

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Merlin, T., Bashi, M.B., Devi, S.J.R. et al. Evaluation of the Anti-Inflammatory Activity of Kokilaksham kashayam on Lipopolysaccharide Stimulated RAW 264.7 Macrophages. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 92, 807–815 (2022). https://doi.org/10.1007/s40011-022-01370-2

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  • DOI: https://doi.org/10.1007/s40011-022-01370-2

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