Withaferin A reduces pulmonary eosinophilia and IL-25 production in a mouse model of allergic airways disease – PubMed Black Hawk Supplements
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Several studies report that ashwagandha, a traditional Ayurvedic supplement, has anti-inflammatory properties. Type 2 (T2) asthma is characterized by eosinophilic airway inflammation. We hypothesized that allergen-induced eosinophilic airway inflammation in mice would be reduced following administration of Withaferin A (WFA), the primary active phytochemical in Ashwagandha. C57BL/6J mice were given 10 total intra-peritoneal injections of 2 mg/kg WFA or vehicle control, concurrent with 6 total…
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Withaferin A reduces pulmonary eosinophilia and IL-25 production in a mouse model of allergic airways disease
Kevin Agner et al. bioRxiv. .
Abstract
Several studies report that ashwagandha, a traditional Ayurvedic supplement, has anti-inflammatory properties. Type 2 (T2) asthma is characterized by eosinophilic airway inflammation. We hypothesized that allergen-induced eosinophilic airway inflammation in mice would be reduced following administration of Withaferin A (WFA), the primary active phytochemical in Ashwagandha. C57BL/6J mice were given 10 total intra-peritoneal injections of 2 mg/kg WFA or vehicle control, concurrent with 6 total intranasal administrations of 50 μg house dust mite extract (HDM) or saline control over 2 weeks. We observed that treatment with WFA reduced allergen-induced peribronchial inflammation and airway eosinophil counts compared to mice treated with controls. In addition, we observed that treatment with WFA reduced lung levels of interleukin-25 (IL-25) but increased lung gene expression levels of its co-receptor, Il17ra, in HDM-challenged mice compared to HDM-challenged mice that received the vehicle control. This study pinpoints a potential mechanism by which WFA modulates allergen-induced airway eosinophilia via the IL-25 signaling pathway. Future studies will investigate the effects of WFA administration on lung eosinophilia and IL-25 signaling in the context of chronic allergen-challenge.
Conflict of interest statement
Declaration of Interest Statement The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Figures

Airway eosinophilic inflammation is reduced with WFA in HDM-challenged mice. (A) Schematic diagram of the mouse model of acute intranasal house dust mite allergen (HDM)-induced allergic airways disease combined with intraperitoneal Withaferin A (WFA) or vehicle control (VC) injection. Control mice were challenged with intranasal phosphate buffered saline (PBS). Bronchoalveolar lavage fluid (BALF) and lung tissue harvest occurred 24 h following the last administrations of HDM/PBS and WFA/VC. Created with
. (B) Representative images of H&E-stained lung sections from all 4 treatment groups. (C) Mean H&E scores ± SEM depicting depth of peribronchial staining for n=2 mice per treatment group. (D) BALF differential cell counts in HDM-challenged mice treated with VC or WFA (n = 2–4 mice per group). Data are presented as mean percentage of total BALF cells ± SEM. **p<0.005 for eosinophils in BALF of both treatment groups in HDM-challenged mice.

In lung tissue of HDM-challenged mice, WFA reduces IL-25 and increases lung Il17ra mRNA expression. (A) IL-33 was measured in whole lung homogenates using ELISA. Data are presented as mean pg/mL IL-33 ± SEM. (n = 3 mice per group). *p<0.050 in HDM-challenged mice compared to saline-challenged mice for both VC and WFA treatment groups. (B) IL-25 was measured in whole lung homogenates using ELISA. Data are presented as mean pg/mL IL-25 ± SEM. *p<0.050 in VC-treated vs WFA-treated HDM-challenged mice (n = 3 mice per group). *p<0.050 in WFA-treated mice comparing saline-challenge to HDM-challenge. (C and D) Il17ra and Il17rb mRNA expression was measured in whole lung RNA using RT-PCR. Data are presented as fold change in Il17ra (C) or Il17rb (D) mRNA expression ± SEM relative to Gapdh mRNA expression and to the mean of the saline-challenged VC-treated mice. (n = 2–3 mice per group). *p<0.050 Il17ra mRNA expression in WFA-treated vs VC-treated HDM-challenged mice.
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