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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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Study Design and Modulation of Arthritis Score by Ashwashila. (A) Male Balb/c mice of 6–8 weeks’ age were injected intraperitoneally with 1.5 mg/mouse dose of anti-collagen antibody (C-Ab) cocktail and 50 µg/mouse of bacterial lipopolysaccharide (LPS). All the disease and treatment group animals were selected randomly from the C-ab induced arthritis (CAIA) animals. The normal (NC) and disease control (DC) animals were treated with sodium carboxymethyl cellulose (Na-CMC) while the treatment group CAIA mice were treated with 353 mg/kg oral dose of ASHW and 0.38 mg/kg oral dose of MTX given every alternate day for two weeks. Physical and clinical parameters of the animals were measured every day throughout the experimental duration. (B) Arthritis score showed an increase in the CAIA animals indicating the onset of RA. Treatment of the CAIA mice with ASHW and MTX significantly reduced the arthritis score indicating a reduction in pedal swellings. (C) Anti-arthritic activity analysis based on the arthritis score showed similar efficacy for both ASHW and MTX except on day 16. Results represent 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 the statistical difference in comparison to MTX (p-value * ≤ 0.05; ** ≤ 0.01).