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. 2019 May 29;9:8025. doi: 10.1038/s41598-019-44485-9

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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In-vitro Modulation of Pro-Inflammatory Cytokines by Ashwashila. (A) THP-1 cells treated with varying concentration of the Ashwashila (ASHW) between 0–25 mg/mL induced minor toxicity at dose of ≥12.5 mg/mL. ASHW concentrations to cause 20% and 50% inhibitions were found at 18.83 mg/mL and 42.29 mg/mL, respectively. Pro-inflammatory responses in the endotoxin lipopolysaccharide (LPS) treated THP-1 cells showed stimulated release of the pro-inflammatory cytokines (B) IL-1β, (C) IL-6 and (D) TNF-α. Treatment of the THP-1 cells with varying concentrations of the Ashwashila (ASHW) inhibited the production of the pro-inflammatory cytokines in a dose-dependent manner. (E) Luciferase NFκB reporter gene vector transfected THP-1 cells were found to express high quantity of NFκB proteins, when stimulated with LPS. This was reduced in a dose-dependent manner in the cells treated with ASHW up to the tested concentration of 10 mg/mL. Values in the results are Mean ± SEM. A one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison t-test was used to calculate the statistical difference. Student unpaired t-test was used to calculate statistical difference in comparison to MTX (p-value # ≤ 0.01; ## ≤ 0.001; * ≤ 0.01; ** ≤ 0.001).