Red Clover Supplementation Alleviates Neurological Deficits in Estrogen Deficient Rats Exposed to Chronic Unpredictable Mild Stress

Estrogen deficiency after menopause contributes to various neurological disorders, including stress, anxiety, depression, and memory impairment. Hormone replacement therapy (HRT) is commonly used to mitigate menopausal symptoms, but its use is associated with significant adverse effects. As a result, phytoestrogens, plant-derived estrogens structurally similar to HRTs, are preferred alternatives due to their lack of side effects associated with synthetic HRTs. Among these phytoestrogens, red clover (RC) has emerged as a potent medicinal herb used for the treatment of menopausal symptoms. Thus, the aim of the current study was to evaluate the effects of RC on neurological disorders in estrogen-deficient rats subjected to chronic unpredictable mild stress (CUMS). Ovariectomy (OVX) was performed to induce estrogen deficiency, a condition that closely mimics menopause in females. CUMS, a model of chronic stress, was employed to mimic the stress and anxiety that commonly accompany menopause. Significant changes in physiological, neurobehavioral, biochemical, molecular, and histopathological alterations in the brain hippocampal region were observed in OVX, CUMS, and OVX + CUMS group rats, indicating enhanced neuronal deficits compared with control group rats. Treatment with RC supplementation, 17-β estradiol (E2), and fluoxetine (Flx) significantly restored the pathological alterations caused by both CUMS and estrogen deficiency toward normal. E2 and Flx were included in the study to serve as established treatments for postmenopausal symptoms and stress-related disorders, providing a basis for comparison with RC. In conclusion, our study demonstrated the immense potential of RC in alleviating neurological disorders associated with estrogen deficiency and chronic stress.