With growing interest in the role of the gut microbiome in the pathogenesis of menopausal disorders, probiotic supplementation is emerging as a novel therapeutic strategy for postmenopausal women. In this study, we investigated the protective effects of several probiotic strains against menopausal symptoms using an ovariectomized (OVX) mouse model fed a high-fat diet, in comparison with the effects of 17β-estradiol (E2) replacement. Among the eight tested probiotic strains, we selected four (Limosilactobacillus fermentum MG901, Lactiplantibacillus plantarum MG4604, Ligilactobacillus salivarius MG4679, and Lacticaseibacillus reuteri MG5463) that showed superior estrogen-like activity in an uterotrophic assay. We then examined their effects in OVX mice. The probiotic MG4679 was tested in combination with MG5463. During the 12-week experimental period, oral administration of the four probiotics reduced blood glucose and body temperature and ameliorated depression-like behavior in OVX mice. In the Barnes maze test, MG901 and MG4604 improved the spatial memory deficits observed in OVX mice. Micro-CT analysis of mouse femurs revealed that MG5463 prevented the OVX-induced reduction in cortical bone mineral density and bone volume fraction. Importantly, similar to E2 replacement, most of the test probiotics increased Akt phosphorylation in skeletal muscle, estrogen receptor-α (ER-α) protein expression in the liver, and ERK phosphorylation in the brain of OVX mice. However, unlike E2 treatment, all probiotics did not affect uterine weight, uterine ER-α expression, and serum E2 concentration. These results indicated that the probiotics used in this study exhibited protective effects against menopausal symptoms, partly through estrogen-mimetic action, without unfavorable uterine stimulation in OVX mice.
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