Traditionally, hyperthyroid-associated osteoporosis has been considered to be the result of increased thyroid hormone levels. The pathogenesis of hyperthyroid-associated osteoporosis remains unclear. Thyroid stimulating hormone receptor (TSHR) is closely associated with osteoporosis. Our study aimed to explore the role of TSHR and its upstream microRNA (miRNA) in hyperthyroid-associated osteoporosis. Bioinformatics analysis (starBase and Targetscan) and a wide range of experiments including reverse-transcription quantitative polymerase chain reaction, luciferase reporter, western blot analysis of osteogenic differentiation markers including OSX, OCN, ALP, OPN, and COL1, hematoxylin and eosin staining, Alizarin Red staining assays were used to explore the function and mechanism of TSHR in hyperthyroid-associated osteoporosis. First, we observed that TSHR was downregulated in bone marrow mesenchymal stem cells (BMSCs) isolated from rats after culture in osteogenic medium for 7 days. Functionally, overexpression of TSHR accelerates BMSC osteogenic differentiation. Mechanistically, we predicted four potential miRNAs for TSHR. MiR-577 was validated to bind with TSHR. Rescue assays showed that miR-577 overexpression inhibited BMSC osteogenic differentiation via targeting TSHR. In vivo experiments showed that miR-577 aggravated bone loss and bone remodeling and our data showed that it is achieved by targeting TSHR in hyperthyroid-associated osteoporosis. This finding may deep our understanding of the pathogenesis of hyperthyroid-associated osteoporosis.