The effect and mechanism of salvianolic acid B on reproductive function in mice with ovarian aging

doi:10.11958/20252197

Abstract

Abstract:

Objective To investigate the effect and potential mechanism of salvianolic acid B (SALB) extracted from salvia miltiorrhiza in ameliorating the aging process by modulating ovarian antioxidant capacity. Methods (1) In vitro experiments: a hydrogen peroxide (H₂O₂)-induced senescence model was established using human ovarian granulosa cell line (KGN), and the effective concentration of H₂O₂ was determined. KGN cells were treated with SALB at varying concentrations and durations. Cell viability was assessed via CCK-8 assay to determine the optimal concentration and treatment time of SALB. Treated KGN cells were divided into the control group (H₂O₂) and the experimental group (H₂O₂ + SALB). Levels of reactive oxygen species (ROS) and malondialdehyde (MDA) were measured. In vivo experiments: forty 9-month-old female mice were randomly assigned to the aged group and the treatment group (n=20 each), while an additional 20 2-month-old mice were used as the young group. The treatment group received intraperitoneal injections of SALB at 40 mg/kg. The young group and the aged group received equivalent doses of normal saline via intraperitoneal injection. Ovarian reserve was evaluated by counting antral follicles via hematoxylin and eosin (HE) staining. Serum levels of estradiol (E2) and follicle-stimulating hormone (FSH) were measured using enzyme-linked immunosorbent assay (ELISA). The estrous cycle was monitored via vaginal smears. Results (1) Treatment with 300 μmol/L H₂O₂ for 12 hours significantly reduced cell viability, inhibited proliferation and induced senescence in KGN cells. Treatment with 20 μmol/L SALB for 12 hours began to show effects, effectively counteracting H₂O₂-induced senescence in KGN cells and significantly reducing intracellular ROS and MDA levels (P<0.05). (2) In vivo results showed that, compared to the young group mice, mice of the aged group exhibited a significant reduction in antral follicle count and serum E2 levels, along with elevated FSH levels (P<0.05), and marked disruption of the estrous cycle. In contrast, mice in the treatment group demonstrated increased antral follicle counts and E2 levels, as well as reduced FSH levels compared to the aged group (P<0.05). There were no significant differences in these indicators between the treatment group and the young group. Moreover, the estrous cycle exhibited regular patterns in the treatment group. Conclusion SALB may exert antioxidant efficacy by scavenging ROS and inhibiting lipid peroxidation, thereby effectively alleviating senescence in ovarian granulosa cells, restoring ovarian reserve, normalizing estrous cycle irregularities, and ameliorating hormonal imbalances.

Key words: Salvianolic acid B, ovarian diseases, cellular senescence, estrous cycle, antioxidation, hormones, fertility

CLC Number:

R711.75，R339.221

Figures/Tables 7

References 20

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组别	MDA/（mmol/g）	ROS/（RU）
H₂O₂组	11.89±2.30	4 206±324
H₂O₂+SALB组	4.83±1.20	2 107±186
t	3.760^＊	3.972^＊＊

组别	MDA/（mmol/g）	ROS/（RU）
H₂O₂组	11.89±2.30	4 206±324
H₂O₂+SALB组	4.83±1.20	2 107±186
t	3.760^＊	3.972^＊＊

组别	窦状卵泡/个	E2/（ng/L）	FSH/（ng/L）
年轻组	9.40±1.14	101.95±6.36	5.35±0.69
老年组	2.80±1.48^a	56.13±6.29^a	13.63±1.56^a
治疗组	6.20±1.10^b	86.81±10.20^b	6.32±1.13^b
F	4.123^＊＊	5.121^＊＊	7.702^＊＊

组别	窦状卵泡/个	E2/（ng/L）	FSH/（ng/L）
年轻组	9.40±1.14	101.95±6.36	5.35±0.69
老年组	2.80±1.48^a	56.13±6.29^a	13.63±1.56^a
治疗组	6.20±1.10^b	86.81±10.20^b	6.32±1.13^b
F	4.123^＊＊	5.121^＊＊	7.702^＊＊

组别	发情间期	发情前期	发情期	发情后期
年轻组	25.24±1.04	24.78±2.36	25.22±4.39	24.76±3.61
老年组	31.94±3.48^a	16.13±4.39^a	36.36±1.16^a	15.57±1.16^a
治疗组	22.68±2.10^b	23.32±5.20^b	26.32±2.13^b	27.68±4.13^b
F	5.163^＊	4.221^＊	5.432^＊	7.221^＊