Efek Fitoestrogen Red Clover (Trifolium Pratense) terhadap Modulasi Interleukin-1β, Interleukin-10, Tumor Necrosis Factor-Α Dan Proses Neurogenesis pada Tikus Galur Sprague Dawley Model Stroke Iskemik

Abstract

Introduction: Stroke occurs due to focal or global disturbances of brain function caused by vascular disorders. However, pharmacological approaches to treat ischemic stroke remain limited. Red clover ( Trifolium pratense ) leaf extract, an isoflavone-containing compound, has potential neuroprotective effects and may play a role in neurogenesis. A reduction in pro-inflammatory mediators and an increase in anti-inflammatory mediators indicate improvement in brain tissue following ischemic injury. This study aims to evaluate the effects of red clover (Trifolium pratense) extract on the modulation of IL-1β, IL-10, TNF-α, and neurogenesis in Sprague–Dawley rats with an ischemic stroke model. Methods: A total of 35 Sprague–Dawley rats with an ischemic stroke model were used in this in-vivo experimental study and randomized into seven groups. The ischemic stroke model was created by unilateral occlusion of the carotid artery, including the left internal and external carotid arteries. A 100% phytoestrogen solution with a 1% concentration at a dose of 40 mg/kg BW was administered. Serum levels of IL-1β, IL-10, and TNF-α were measured using ELISA. Viable neurons and astrocytes in brain tissue were evaluated through histopathological examination. Results: Statistical analysis revealed significant differences in IL-1β levels between the control group and stroke-model rats with and without phytoestrogen administration at both 1 and 7 days (p = 0.012 vs p < 0.001). Similarly, IL-10 levels differed significantly between the 1-day and 7-day groups (p < 0.001 vs p = 0.007). For TNF-α, significant differences were found only in the 7-day groups (p = 0.233 vs p < 0.001). Additionally, a significant difference was observed in the mean number of astrocytes across groups (p < 0.001), whereas no significant difference was found in the mean number of viable neurons (p = 0.134). Conclusion: Red clover (Trifolium pratense) extract appears to reduce proinflammatory cytokines (IL-1β and TNF-α) and increase anti-inflammatory cytokines (IL-10). However, its effect on neurogenesis was not evident in this study. The observed number of astrocytes should be interpreted as preliminary findings based on morphological assessment