Assessment of Hematologic-Enhancing Potentials and Toxicity of Aqueous Fraction of Justicia secunda (Vahl) Leaf on Phenylhydrazine-Induced Anaemic Albino Rats

Abstract

Anaemia remains a major health challenge, and the search for affordable, accessible, and biologically effective plant-based therapies has increased interest in medicinal plants traditionally used for blood-related disorders. This study evaluated the blood-enhancing potential and toxicity profile of the aqueous fraction of Justicia secunda Vahl leaf extract in phenylhydrazine (PHZ)-induced anaemic albino rats. The aqueous fraction was subjected to gas chromatography–mass spectrometry (GC–MS) analysis to characterise its phytochemical constituents. Acute toxicity was evaluated using graded oral doses of 10–5000 mg/kg body weight. Anaemia was induced with phenylhydrazine, after which animals were allocated to normal control, standard drug-treated, untreated anaemic, and J. secunda-treated groups receiving 200, 400, or 600 mg/kg body weight of the aqueous fraction. Haematological indices, including packed cell volume, haemoglobin concentration, red blood cell count and erythrocyte indices, were assessed together with total and differential leukocyte counts. Serum iron, total iron-binding capacity and unsaturated iron-binding capacity were also determined. In addition, malondialdehyde (MDA), superoxide dismutase (SOD), catalase, reduced glutathione (GSH), and myeloperoxidase (MPO) were evaluated as indices of oxidative stress and systemic inflammation. GC–MS analysis identified several classes of compounds, with aliphatic hydrocarbons (43.35%), fatty acids (26.08%), aldehydes (13.69%), phenolic compounds (14.82%), and aromatic halogenated compounds (0.31%) predominating. No mortality or overt signs of toxicity were observed at doses up to 5000 mg/kg, indicating a relatively wide acute oral safety margin. Administration of the aqueous fraction was associated with improvement in haematological parameters and modulation of iron-related, antioxidant and inflammatory biomarkers in PHZ-induced anaemic rats. The findings suggest that J. secunda leaf aqueous fraction possesses blood-enhancing and antioxidant properties and may support recovery from chemically induced haemolytic anaemia. However, further mechanistic investigations, subchronic toxicity studies, and clinical investigations are warranted to establish its therapeutic safety and efficacy.