丹酚酸B对卵巢衰老小鼠生殖功能的影响及机制探讨

doi:10.11958/20252197

天津医药 ›› 2026, Vol. 54 ›› Issue (2): 117-121.doi: 10.11958/20252197

• 专题研究·生殖系统疾病 • 上一篇下一篇

丹酚酸B对卵巢衰老小鼠生殖功能的影响及机制探讨

谷梦青(), 李佳林, 于洋()

北京大学第三医院生殖医学中心（邮编100191）

收稿日期:2025-06-05 修回日期:2025-09-03 出版日期:2026-02-15 发布日期:2026-02-12
通讯作者: 于洋 E-mail:gumengqing163@163.com;yuyang5012@hotmail.com
作者简介:谷梦青（1997），女，博士在读，主要从事女性生育力改善方面研究。E-mail：gumengqing163@163.com
基金资助:
国家重点研发计划项目(2024YFC2706600);国家自然科学基金资助项目(82225019);国家自然科学基金资助项目(82192873)

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

GU Mengqing(), LI Jialin, YU Yang()

Center for Reproductive Medicine, Peking University Third Hospital, Beijing 100191, China

Received:2025-06-05 Revised:2025-09-03 Published:2026-02-15 Online:2026-02-12
Contact: YU Yang E-mail:gumengqing163@163.com;yuyang5012@hotmail.com

谷梦青, 李佳林, 于洋. 丹酚酸B对卵巢衰老小鼠生殖功能的影响及机制探讨[J]. 天津医药, 2026, 54(2): 117-121.

GU Mengqing, LI Jialin, YU Yang. The effect and mechanism of salvianolic acid B on reproductive function in mice with ovarian aging[J]. Tianjin Medical Journal, 2026, 54(2): 117-121.

摘要/Abstract

摘要：

目的探究丹参提取物丹酚酸B（SALB）通过调节卵巢抗氧化能力改善衰老进程的作用及可能机制。方法（1）细胞实验。H₂O₂构建人卵巢颗粒细胞系（KGN）衰老细胞模型并进行有效作用浓度筛选。不同浓度和不同作用时长的SALB处理KGN细胞，CCK8染色检测细胞活力并确定SALB有效浓度及作用时间。KGN细胞经不同处理分为对照（H₂O₂）组和实验（H₂O₂+SALB）组，检测活性氧（ROS）和丙二醛（MDA）水平。（2）动物实验。40只9月龄小鼠均分为老年组和治疗组，每组20只；20只2月龄小鼠为年轻组。治疗组小鼠给予40 mg/kg的SALB腹腔注射治疗。年轻组和老年组仅予以适应性喂养，腹腔注射与治疗组等剂量的生理盐水。HE染色计数窦状卵泡评估卵巢储备；酶联免疫吸附试验（ELISA）检测血清雌二醇（E2）、卵泡刺激素（FSH）的水平；阴道涂片统计发情周期。结果（1）细胞实验。用300 μmol/L的H₂O₂处理12 h可以显著降低细胞活力、抑制细胞增殖，使KGN细胞处于衰老状态。20 μmol/L的SALB处理12 h开始有效果，可有效降低H₂O₂诱导的KGN的衰老，降低衰老细胞内ROS和MDA的水平（P<0.05）。（2）动物实验。与年经组小鼠相比，老年组小鼠卵巢组织窦状卵泡数和E2水平降低、FSH水平升高（P<0.05），发情周期明显异常；与老年组小鼠相比，治疗组小鼠卵巢组织窦状卵泡数和E2水平升高、FSH水平降低（P<0.05），且治疗组指标与年经组比较，差异无统计学意义，发情周期呈规律性变化。结论 SALB可能通过清除ROS、抑制脂质过氧化发挥抗氧化效能，有效改善卵巢颗粒细胞衰老并恢复卵巢储备、调整发情周期及激素异常。

关键词: 丹酚酸B, 卵巢疾病, 细胞衰老, 发情周期, 抗氧化, 激素, 生育力

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

中图分类号:

R711.75，R339.221

图/表 7

图1 H2O2处理后细胞活力检测 A—F分别为空白组，100、200、300、400和500 μmol/L H2O2组；n=3，F=90.163，P<0.05；a与空白组比较，P<0.05。

Fig. 1 Cell viability assay after H2O2 treatment

图2 H2O2处理后各组细胞增殖检测（×100）

Fig.2 Cell proliferation assay after H2O2 treatment (×100)

图3 SALB处理6 h、12 h及24 h后衰老KGN细胞活力分析 A—F分别为空白组，H2O2组及5、10、20、40和60 μmol/L SALB+ H2O2组；F6 h=198.231，F12 h=256.476，F24 h=267.452，均P<0.05；a与空白组比较，b与H2O2组比较，P<0.05。

Fig. 3 Cell viability analysis of senescent KGN after SALB treatment for 6 h, 12 h and 24 h

表1 2组细胞中MDA和ROS水平比较（n=3,$\bar{x}±s$）

Tab.1 Comparison of MDA and ROS levels between two groups of cells

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^＊＊

图4 各组小鼠卵巢组织窦状卵泡数表现（HE，×50）

Fig.4 Performance of ovarian follicle numbers in each group of mice (HE，×50)

表2 3组小鼠血清激素水平以及卵巢中窦状卵泡数目比较（n=20,$\bar{x}±s$）

Tab.2 Comparison of serum hormone levels and antral follicle counts in ovaries between the three mouse groups

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^＊＊

表3 3组小鼠发情周期各阶段占比比较（n=20,%,$\bar{x}±s$）

Tab.3 Percentage distribution of each stage in the estrous cycle across three groups of mice

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^＊

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丹酚酸B对卵巢衰老小鼠生殖功能的影响及机制探讨

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

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