Efficacy of standardised Orthosiphon stamineus extract (C5EOS5050ESA) in managing Cytokine storm in mouse model of Malaria infection using Plasmodium berghei anka

Malaria is a prevalent global health issue, marked by excessive immune responses which initiate and amplify pathophysiological processes, leading to "cytokine storm" and multi-organ damage. This study investigated the therapeutic potential of Orthosiphon stamineus extract (OSE) in murine model of malaria using Plasmodium berghei ANKA. The effects of OSE treatment were evaluated on cytokine production, histopathological changes and the overall course of infection. Infection was established by intraperitoneal inoculation of male ICR mice with 0.1 mL of 1 x 10⁷ parasitized red blood cells (PRBCs). Mice were experimentally grouped into five (n=8 in each) which include the control uninfected group, the malarial group treated with Tween80 (vehicle), and malarial groups treated with either 50, 200 or 400 mg/kg/day of OSE doses. Basic parameters including body weight, body temperature and parasitaemia were monitored daily throughout the course of the infection. Major organs (brain, liver, spleen, lungs, and kidneys) were harvested from the mice for determination of the effects of treatment on histopathological conditions during the infection. Plasma and serum were prepared from blood collected from the mice for cytokine analysis. IFN-α and IFN-β in the plasma of the mice were quantified using Enzyme-linked immunosorbent Assay (ELISA), while IL-2, IL-4, IL-6, IL-10, IL-17A, TNF-α, and IFN-γ was determined in the serum via Cytometric Bead Array (CBA) analysis. Results showed that treatment with OSE improved the histopathological conditions across major organs. Sequestration and hemozoin deposition were reduced in OSE-treated malarial groups as compared to the Tween80-treated malarial group. Treatment with OSE also caused a significant reduction in TNF-α, IFN-γ, IL-2, IL-4, IL-6, and IL-10 levels in a dose-dependent manner as compared to malaria treated with Tween80, indicating potential inhibitory and immunomodulatory effects against excessive cytokine release during malaria infection. OSE treatment, particularly at a dose of 400 mg/kg/day, significantly enhanced survival rates (p < 0.001) and reduced parasitaemia levels (p < 0.001) throughout the course of infection compared to the malarial group treated with Tween80. Decreased parasitaemia correlated with significantly elevated haemoglobin levels observed in all OSE-treated groups on day 5 post-inoculation (p < 0.05 for 50 mg/kg, p < 0.001 for 200 and 400 mg/kg). Overall findings in this study suggest that the standardised OSE may serve as a novel treatment strategy targeting inflammatory and immune mediators during malaria infection. The findings offer promising solution in mitigating inflammatory responses and improving survival outcomes in malaria.

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