Pinto Bean and Resistant Starch Supplementation Modulate Gut Microbiota and Metabolic Outcomes in Beta-Glucuronidase-Inhibited Estrogen-Deficient Mice
This study investigates the effects of pinto bean (PB) and resistant starch (RS3) supplementation on gut microbiota composition and metabolic outcomes in estrogen-deficient mice with targeted inhibition of gut bacterial beta-glucuronidase (GUS). Female C57BL/6 mice were rendered estrogen-deficient through daily injections of 4-vinylcyclohexene diepoxide (VCD) for 30 days. After confirmation of ovarian failure, mice were randomized into dietary and treatment groups and administered a commercially available beta-glucuronidase inhibitor (1-((6,8-Dimethyl-2-oxo-1,2-dihydroquinolin-3-yl)methyl)-3-(4-ethoxyphenyl)-1-(2-hydroxyethyl)thiourea, Sigma-Aldrich, St. Louis, MO). This trisubstituted thiourea compound is a potent, uncompetitive, reversible, and targeted inhibitor of gut bacterial beta-glucuronidase.Mice were maintained on their respective diets with or without beta-glucuronidase inhibitor for 14 weeks. Cecal contents were collected at the endpoint, and DNA was extracted for 16S rRNA gene sequencing using the Illumina platform. Sequencing was performed with forward and reverse reads for each sample.The study includes six experimental groups, each with eight biological replicates:ConVeh: Control diet, vehicleConInh: Control diet, beta-glucuronidase inhibitorPBVeh: Pinto bean diet, vehiclePBinh: Pinto bean diet, beta-glucuronidase inhibitorRS3Veh: Resistant starch diet, vehicleRS3Inh: Resistant starch diet, beta-glucuronidase inhibitorThe resulting 16S rRNA sequencing data provide a resource to examine how dietary interventions and gut bacterial beta-glucuronidase inhibition interact to modulate gut microbial composition and associated metabolic outcomes in estrogen-deficient conditions.