Bushen Zhuluan Decoction Ameliorates Diminished Ovarian Reserve in Rats via the SIRT1/HIF-1α Pathway

Diminished ovarian reserve (DOR) is a major cause of female infertility. In recent years, traditional Chinese medicine has displayed unique ameliorating effects on DOR. This study aims to investigate the therapeutic efficacy and molecular mechanisms of Bushen Zhuluan Decoction (BSZLD) in DOR. A rat model of DOR was established and treated with either BSZLD or Progynova (positive control). Estrous cycle monitoring, ovarian index calculation, hematoxylin-eosin staining, and enzyme-linked immunosorbent assay were performed to assess ovarian reserve function. TUNEL staining, immunohistochemistry, and Western blotting were conducted to evaluate ovarian cell Apoptosis. To elucidate the pharmacological basis of BSZLD, ultra-high-performance liquid chromatography-tandem mass spectrometry was applied. Network pharmacology analysis, machine learning algorithms, and molecular docking were integrated to predict key active compounds and pathways. The role of the Sirtuin 1 (SIRT1) and its downstream target hypoxia-inducible factor 1alpha (HIF-1α) signaling in BSZLD-mediated effects was verified using short hairpin RNA (shRNA) lentiviral-mediated interference. BSZLD significantly restored estrous cycle disturbances, increased ovarian index and follicle counts, and reduced atretic follicles and ovarian cell Apoptosis in DOR rats. Serum anti-Müllerian hormone and estradiol levels were elevated, while follicle-stimulating hormone and luteinizing hormone levels were reduced. The SIRT1/HIF-1α signaling pathway was a core regulatory axis for BSZLD action. BSZLD markedly upregulated SIRT1 expression while suppressing HIF-1α expression. Silencing SIRT1 using shRNA reduced SIRT1 levels, elevated HIF-1α expression, and significantly attenuated the therapeutic effects of BSZLD. BSZLD attenuates DOR in rats by activating the SIRT1/HIF-1α signaling, thereby suppressing ovarian cell Apoptosis and restoring ovarian reserve function.