Safety and Metabolic Effects of Withania somnifera Seed Extract and Powder in Rats: Insights From DPP‐4 Molecular Docking

ABSTRACT Withania somnifera seeds remain understudied compared with roots despite increasing nutraceutical use. This study evaluated safety, metabolic effects, and potential DPP‐4 activity of seed extract (WSS‐E) and powder (WSS‐P) in rats. Twenty male Wistar rats were assigned to five groups ( n = 4/group): control, 0.25% WSS‐E, 0.5% WSS‐E, 0.5% WSS‐P, and 1% WSS‐P for 56 days. Biochemical, hematological, and histopathological outcomes were assessed, whereas major seed phytochemical–DPP‐4 interactions were examined by molecular docking. Data were analyzed by ANOVA and Duncan's test ( p < 0.05). The 0.5% WSS‐E group showed the strongest metabolic response. At Day 56, glucose (85.73 vs. 105.69 mg/dL), triglycerides (78.40 vs. 108.84 mg/dL), total cholesterol (90.55 vs. 104.03 mg/dL), LDL (37.69 vs. 48.53 mg/dL), VLDL (15.70 vs. 21.78 mg/dL), and atherogenic index (2.13 vs. 3.24) were significantly lower than control, whereas HDL was higher (37.19 vs. 33.76 mg/dL; p < 0.05). Creatinine and urea were also significantly lower than control. No overt hepatic, renal, hematological, or histopathological toxicity was observed. Withanone showed strongest predicted DPP‐4 binding (−10.0 kcal/mol), comparable to sitagliptin (−8.4 kcal/mol). Thus, 0.5% WSS‐E provided the most favorable metabolic profile without overt toxicity; long‐term safety and mechanistic studies are warranted.