Objective:To observe the effect of the Yushen Yangluan decoction on ovarian reserve function in mice with diminished ovarian reserve (DOR) and to explore its mechanism of action based on the hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor A (VEGFA) signaling pathway.Methods:Mechanism analysis was conducted using artificial intelligence-based network pharmacology. Fifty 6-week-old female C57BL/6 mice were randomly divided into five groups according to body weight, with 10 mice in each group. The groups were as follows: control group, model group, low-dose Traditional Chinese Medicine (TCM) group, medium-dose TCM group, and high-dose TCM group. Hematoxylin-eosin staining was used to observe the number of antral follicles in the ovaries, enzyme-linked immunosorbent assay was employed to detect serum hormone levels, and TdT-mediated dUTP nick end labeling (TUNEL) was used to assess apoptosis in ovarian granulosa cells. Immunohistochemistry (IHC) was performed to detect ovarian expression of HIF-1α, and Western blotting was used to measure the protein expressional levels of HIF-1α, VEGFA, and B-cell lymphoma-2 (BCL2) in the ovaries.Results:After treating DOR model mice with the Yushen Yangluan decoction at low, medium, and high dosages, the ovarian antral follicle count of the mice in the TCM groups was all higher than that in the model group ( P=0.024, P<0.001, P<0.001), the follicle-simulating hormone level was lower than that in the model group ( P=0.003, P<0.001, P<0.001), and the serum anti-Müllerian hormone level in the medium-and high-dose TCM groups was higher than that in the model group ( P=0.001, P<0.001). Network pharmacology and molecular docking suggested that the formula may influence the HIF signaling pathway through core targets such as HIF-1α, BCL2 and VEGFA. TUNEL assays indicated that the apoptosis rate of ovarian granulosa cells in the low-, medium-, and high-dose TCM groups was significantly lower than that in the model group ( P=0.017, P=0.003, P<0.001). Immunohistochemistry results showed that the expression level of HIF-1α in the granulosa cells of the follicles in the low-, medium- and high-dose TCM groups was significantly higher than that in the model group ( P=0.008, P<0.001, P<0.001). Western blotting analysis indicated that the protein expression level of HIF-1α in medium-, and high-dose TCM groups was significantly higher than that in the model group ( P=0.004, P<0.001), and the protein expression levels of VEGFA and BCL2 in low-, medium-, and high-dose TCM groups were significantly higher than those in the model group ( P=0.003, P=0.002, P=0.001; P=0.029, P=0.007, P=0.001). Conclusion:The Yushen Yangluan decoction may exert its therapeutic effect by inhibiting apoptosis of ovarian granulosa cells through the HIF-1α/VEGFA signaling pathway.

Objective:To observe the effect of the Yushen Yangluan decoction on ovarian reserve function in mice with diminished ovarian reserve (DOR) and to explore its mechanism of action based on the hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor A (VEGFA) signaling pathway.Methods:Mechanism analysis was conducted using artificial intelligence-based network pharmacology. Fifty 6-week-old female C57BL/6 mice were randomly divided into five groups according to body weight, with 10 mice in each group. The groups were as follows: control group, model group, low-dose Traditional Chinese Medicine (TCM) group, medium-dose TCM group, and high-dose TCM group. Hematoxylin-eosin staining was used to observe the number of antral follicles in the ovaries, enzyme-linked immunosorbent assay was employed to detect serum hormone levels, and TdT-mediated dUTP nick end labeling (TUNEL) was used to assess apoptosis in ovarian granulosa cells. Immunohistochemistry (IHC) was performed to detect ovarian expression of HIF-1α, and Western blotting was used to measure the protein expressional levels of HIF-1α, VEGFA, and B-cell lymphoma-2 (BCL2) in the ovaries.Results:After treating DOR model mice with the Yushen Yangluan decoction at low, medium, and high dosages, the ovarian antral follicle count of the mice in the TCM groups was all higher than that in the model group ( P=0.024, P<0.001, P<0.001), the follicle-simulating hormone level was lower than that in the model group ( P=0.003, P<0.001, P<0.001), and the serum anti-Müllerian hormone level in the medium-and high-dose TCM groups was higher than that in the model group ( P=0.001, P<0.001). Network pharmacology and molecular docking suggested that the formula may influence the HIF signaling pathway through core targets such as HIF-1α, BCL2 and VEGFA. TUNEL assays indicated that the apoptosis rate of ovarian granulosa cells in the low-, medium-, and high-dose TCM groups was significantly lower than that in the model group ( P=0.017, P=0.003, P<0.001). Immunohistochemistry results showed that the expression level of HIF-1α in the granulosa cells of the follicles in the low-, medium- and high-dose TCM groups was significantly higher than that in the model group ( P=0.008, P<0.001, P<0.001). Western blotting analysis indicated that the protein expression level of HIF-1α in medium-, and high-dose TCM groups was significantly higher than that in the model group ( P=0.004, P<0.001), and the protein expression levels of VEGFA and BCL2 in low-, medium-, and high-dose TCM groups were significantly higher than those in the model group ( P=0.003, P=0.002, P=0.001; P=0.029, P=0.007, P=0.001). Conclusion:The Yushen Yangluan decoction may exert its therapeutic effect by inhibiting apoptosis of ovarian granulosa cells through the HIF-1α/VEGFA signaling pathway.