Yangjingzhongyu Decoction regulates primordial follicle initiation via LncRNA-Smad1/AMPK dual pathways in diminished ovarian reserve

Targeting the dysregulated initiation of primordial follicles represents a potential therapeutic breakthrough in managing pathological diminished ovarian reserve (DOR). Previous research has demonstrated that Yangjingzhongyu Decoction (YJZYD) promotes the transition from primordial to primary follicles by post-transcriptionally regulating Smad1 protein via the ovary-specific long non-coding RNA (LncRNA) LTCONS-00011173. This effect may be further supported by YJZYD’s ability to ameliorate oxidative stress in the follicular developmental microenvironment through modulation of the AMPK pathway. This study aims to integrate these two mechanistic axes to elucidate the holistic mechanism through which YJZYD modulates the primordial follicle microenvironment. To investigate YJZYD’s regulatory mechanism on the primordial follicle initiating microenvironment in DOR rats. In DOR model rats, primordial and growing follicles significantly decreased (p < 0.05), accompanied by reduced E2 and AMH levels (p < 0.05) and elevated FSH (p < 0.05). Treatment with YJZYD-medicated serum increased counts of primordial/growing follicle counts (p < 0.05), with no significant difference between the high-dose group and the control group (p > 0.05). Additionally, YJZYD reduced FSH, increased E2 and AMH (p < 0.05). It also downregulated LTCONS-00011173 and upregulated Smad1 (p < 0.05). Furthermore, YJZYD elevated SOD activity and ATP content (p < 0.05) and reduced MDA levels (p < 0.05). Moreover, it suppressed PTEN expression, dose-dependently regulated MST and AMH (p < 0.05), and enhanced GDF9 expression at low dose (p < 0.05). Molecular docking identified Apigenin and Curcumin as key monomers targeting AMPK/SIRT1/MAPK pathway. Experimental validation of Apigenin replicated YJZYD’s effects on follicle numbers and hormone levels (p < 0.01). YJZYD alleviates primordial follicle initiation inhibition by targeting the LTCONS-00011173/Smad1 axis, while concurrently improving the follicular microenvironment through AMPK/SIRT1/PGC-1α mediated oxidative stress repair and energy metabolism enhancement. This dual-pathway mechanism restores follicular factor balance and ovarian reserve function in DOR.