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Background: miRNAs are closely related to bone metabolism. Studies have shown that Erxian decoction can improve bone metabolism, possibly achieving this regulatory effect through miRNA targets. Netinfer was used to predict the miRNA targets of Erxian decoction for the treatment of postmenopausal osteoporosis, and the results were validated by clinical trials. Methods: In this study, we identified possible targets of Erxian decoction in osteoporosis by means of network pharmacological analysis and bioinformatic prediction. Fifteen cases of postmenopausal osteoporosis with kidney Yin and Yang deficiency (In traditional Chinese medicine, kidney Yin nourishes and moistens the tissues of the internal organs of the body, while kidney Yang promotes and warms the tissues of the internal organs of the body.) were treated with Erxian decoction for four weeks, and serum bone metabolism indices (P1NP, osteocalcin, and ß-CTX) and miRNA-335-5p expression were measured before and after treatment. Results: The constructed miRNA postmenopausal osteoporosis related gene network of the effective compound of the Erxian decoction has 296 points and 981 edges. The 39 postmenopausal osteoporosis related genes regulated by miRNA-335-5p were enriched in ossification, while the signaling pathways were enriched in rheumatoid arthritis, the Toll signaling pathway, the HIF-1 signaling pathway, and the MAPK signaling pathway. After taking Erxian decoction, the expression of the serum bone formation index (P1NP, osteocalcin) and miRNA-335-5p gene expression levels increased significantly. The alterations in P1NP and osteocalcin were correlated with the changes in miRNA-335-5p. Conclusion: Circulating miRNA-335-5p may serve as an important target of Erxian decoction in the treatment of postmenopausal women. The effect of Erxian decoction on bone formation is significant, but the underlying mechanism requires further investigation. [ABSTRACT FROM AUTHOR]