基于AMPK/SIRT1信号通路探讨补肾健脾方对卵巢储备功能下降大鼠能量代谢的影响*

作者：周 芳1,2，曾 晶1，刘牧涯2，陈嘉盈2，王永连2，尤昭玲1

单位：1.湖南中医药大学第一附属医院，湖南 长沙 410007； 2.湖南中医药大学第一中医临床学院，湖南 长沙 410208

引用：引用：周芳，曾晶，刘牧涯，陈嘉盈，王永连，尤昭玲.基于AMPK/SIRT1信号通路探讨补肾健脾方对卵巢储备功能下降大鼠能量代谢的影响[J].中医药导报,2026,32(5):20-25,68.

DOI：10.13862/j.cn43-1446/r.2026.05.004

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摘要：

目的：从能量代谢角度探讨补肾健脾（BSJP）方调控大鼠卵巢储备功能下降的机制。方法：将50只动情周期正常的雌性SD大鼠随机分成正常组10只和造模组40只。造模组予环磷酰胺75 mg/kg一次性腹腔注射造模。造模成功后随机分成模型组、西药组[芬吗通0.204 mg/（kg·d）]、BSJP等量组[6.43 g/（kg·d）]及BSJP高量组[12.85 g/（kg·d）]，连续给药15 d。酶联免疫吸附试验（ELISA）测定血清中卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、抗米勒管激素（AMH）水平；苏木精-伊红（HE）染色观察大鼠卵巢结构；免疫组化法检测卵巢AMPK蛋白表达量；蛋白质印迹法（Western blotting）及实时定量聚合酶链反应（qPCR）检测卵巢AMPK、SIRT1蛋白及基因表达量。结果：与正常组比较，模型组大鼠动情周期紊乱，FSH、LH水平明显升高（P<0.01），E2、AMH明显降低（P<0.01）；HE染色显示大鼠卵巢结构破坏；ATP水平、AMPK蛋白、SIRT1蛋白及基因表达降低（P<0.01或P<0.05）。与模型组比较，给药干预后各组大鼠FSH、LH水平均明显下降（P<0.01），E2、AMH均明显升高（P<0.01）；卵巢结构均有不同程度的恢复；西药组、BSJP高量组、BSJP等量组大鼠干预后ATP均增多（P<0.01或P<0.05）；BSJP等量组及BSJP高量组大鼠免疫组化检测AMPK蛋白表达升高（P<0.01），西药组AMPK蛋白表达差异无统计学意义（P＞0.05）；Western blotting检测西药组、BSJP高量组及BSJP等量组大鼠AMPK蛋白表达升高（P<0.05或P<0.01），西药组和BSJP高剂量组大鼠AMPK基因表达明显升高（P<0.05或P<0.01），BSJP等量组大鼠AMPK基因表达变化差异无统计学意义（P>0.05），西药组和BSJP高量组大鼠SIRT1蛋白及基因表达量均显著升高（P<0.01），BSJP等量组大鼠SIRT1蛋白表达量、基因表达差异均无统计学意义（P>0.05）。结论：补肾健脾方能改善卵巢储备功能，其可能通过激活AMPK/SIRT1通路，促进AMPK、SIRT1表达，增加ATP合成，改善卵巢能量代谢障碍，从而治疗环磷酰胺对大鼠卵巢储备功能的损伤。

关键词：卵巢储备功能下降；补肾健脾方；AMPK/SIRT1通路；ATP；大鼠

Abstract：

Objective: To explore the mechanism by which the Bushen Jianpi formula (BSJP) regulates diminished ovarian reserve in rats from the perspective of energy metabolism. Method: Totally 50 normal female SD rats were randomly divided into the normal group (n=10) and the modeling group (n=40). The modeling group was given cyclophosphamide (75 mg/kg) by one-time intraperitoneal injection. After successful modeling, the rats were randomly divided into a model group, a Western medicine group [Femoston 0.204 mg/(kg·d)], a BSJP equivalent-dose group [6.43 g/(kg·d)], and a BSJP high-dose group [12.85 g/(kg·d)], and the rats were treated continuously for 15 days. The levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and anti-Muller-tube hormone (AMH) were determined by enzyme linked immunosorbent assay (ELISA). The ovarian structure of rats was observed by HE staining. The expression of AMPK protein in ovary was detected by immunohistochemistry. Ovarian AMPK, SIRT1 protein and gene expression were detected by Western blotting and Real time quantitative polymerase chain reaction (qPCR). Results: Compared with the normal group, the estrous cycle of rats in the model group was disordered, the levels of FSH and LH were significantly increased (P<0.01), and E2 and AMH were significantly decreased (P<0.01). HE staining showed that the ovarian structure of rats was severely damaged in the model group. The levels of ATP, AMPK protein, SIRT1 protein and gene expressions decreased in the model group (P<0.01 or P<0.05). Compared with the model group, after drug administration intervention, the levels of FSH and LH in each group of rats decreased significantly (P<0.01), and E2 and AMH increased significantly (P<0.01). HE staining showed that the ovarian structures all recovered to varying degrees. The ATP of the ovaries increased significantly in the Western medicine group, BSJP equivalent-dose group and BSJP high-dose group (P<0.01 or P<0.05), and immunohistochemical tests showed that the expression of AMPK protein increased in the BSJP equivalent-dose group and BSJP high-dose group (P<0.01), while there was no statistically significant expression of AMPK protein in the Western medicine group (P＞0.05). The expression of AMPK protein, detected by Western blotting, in the Western medicine group, BSJP equivalent-dose group and BSJP high-dose group was significantly increased (P<0.05 or P<0.01). The expression of AMPK gene in the Western medicine group and BSJP high-dose group was significantly increased (P<0.05 or P<0.01). There was no statistically significant change in the expression of AMPK gene in the BSJP equivalent-dose group (P>0.05). The expression levels of SIRT1 protein and gene in the Western medicine group and BSJP high-dose group were significantly increased (P<0.01), while there was no statistically significant change in the expression level of SIRT1 protein in BSJP equivalent-dose group (P>0.05). Conclusion: Bushen Jianpi formula can improve ovarian reserve function, possibly by activating the AMPK/SIRT1 signaling pathway, promoting the expression of AMPK and SIRT1, increasing ATP synthesis, and ameliorating ovarian energy metabolism disorders, thereby treating cyclophosphamide-induced damage to ovarian reserve function in rats.

Key words：decreased ovarian reserve function; Bushen Jianpi formula; AMPK/SIRT1 pathway; ATP; rat

发布时间：2026-05-23

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