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AbstractOne of the prominent complications observed in those undergoing treatment with methotrexate (MTX) is hematological profile alterations which could culminate in severe anemia. In this study, we assessed the hematological profile indices in MXT-treated rats and the effect of leaf extract of Datura stramonium (LEDS) supplementation in MXT-treated rats. Ethanol (98%) was the solvent used in extraction. Animals were divided at random into four groups. Animals in group 1 received normal saline (5 mg/kg) orally and feeding was limitless and did not receive MXT. Animals in group 2 were given LEDS orally (200 mg/kg body weight) for 21 days while group 3 received 20 mg/kg body weight (bw) of MXT on day 18 via the intra-peritoneum without LEDS. Rats in group 4 were given the extract (200 mg/kg bw) and also injected with 20 mg/kg bw of MXT on day 18 of the study via the intra-peritoneum. Serum levels of hemoglobin, red blood cells, packed cell volume, total white blood cells, neutrophils, lymphocytes, and platelets were determined. Rats treated with MXT had notable depletion in hemoglobin, red blood cells, packed cell volume, total white blood cells, neutrophils, and platelets, unlike the control group. Interestingly, LEDS supplementation markedly restored the altered hematological profiles. MXT injection caused hematological dysfunction while co-supplementation with LEDS restored the impaired hematological indices. Therefore, LEDS could be a promising tool in arresting hematological dysfunctions accompanying MXT chemotherapy. However, we advocate for further prospective scrutiny.