| 摘要: |
| ［摘要］ 目的 探讨当归芍药散治疗早发性卵巢功能不全（POI）的作用机制。方法 通过腹腔注射环磷酰胺（CTX）构建POI大鼠模型，设置空白组、模型组、促性腺激素释放激素类似物（GnRHa）组（在造模前第10天和第5天予GnRHa腹腔注射，0.1 mg/kg）和当归芍药散组（在造模后予当归芍药散灌胃，10 mL/kg），每组6只，连续干预14 d。通过HE染色和透射电子显微镜观察大鼠卵巢组织形态学结构。采用酶联免疫吸附试验（ELISA）检测血清卵泡刺激素（FSH）、雌二醇（E2）、抗米勒管激素（AMH）水平。通过Western blot实验和逆转录聚合酶链反应（RT-PCR）实验检测卵巢组织转化生长因子β1（TGFβ1）-Smad2信号通路关键因子表达水平。结果 模型组大鼠血清E2、AMH水平显著下降，FSH水平显著升高；卵巢组织中闭锁卵泡增多，颗粒细胞疏松，细胞器重度水肿，组织间隙紊乱，细胞膜破损，细胞核不规则凹陷，表明卵巢功能受损严重。经当归芍药散、GnRHa干预后，POI模型大鼠血清E2、AMH水平回升，FSH水平下降；卵巢组织内空泡减少，各级卵泡增多，细胞膜及核膜破损与模型组相似，细胞器肿胀减轻。模型组卵巢组织磷酸化转化生长因子β受体1（p-TGFβR1）、磷酸化Smad2（p-Smad2）蛋白表达水平显著升高（P<0.05），TGFβ1、转化生长因子β受体1（TGFβR1）、转化生长因子β受体2（TGFβR2）、Smad2蛋白及mRNA表达水平显著升高（P<0.05），Smad6、Smad7蛋白及mRNA表达水平显著降低（P<0.05），表明TGFβ1-Smad2信号通路过度激活。经当归芍药散干预后，Smad2、p-Smad2蛋白表达水平显著降低（P<0.05），TGFβ1、TGFβR1、TGFβR2蛋白及mRNA表达水平显著降低（P<0.05），Smad6、Smad7蛋白及mRNA表达水平显著升高（P<0.05），各指标变化与GnRHa干预相似。结论 当归芍药散可能通过抑制TGFβ1-Smad2信号通路的异常激活发挥治疗POI的作用。 |
| 关键词: 当归芍药散 早发性卵巢功能不全 TGFβ1-Smad2信号通路 动物实验 |
| DOI：10.3969/j.issn.1674-3806.2025.08.07 |
| 分类号:R 711.75 |
| 基金项目:国家自然科学基金项目（编号：82360949）；海南省自然科学基金高层次人才项目（编号：821RC716）；海南省自然科学基金青年基金项目（编号：821QN0997）；海南省临床医学中心建设项目（编号：琼卫医函［2021］75号） |
|
| Investigation on mechanisms of Danggui Shaoyao Powder in treating premature ovarian insufficiency |
|
WANG Jing, ZHAO Rui, CHEN Yang, ZHANG Xiaojuan, XIA Xiaoyan
|
|
Department of Traditional Chinese Medicine, Hainan Women and Children′s Medical Center, Haikou 571100, China
|
| Abstract: |
| ［Abstract］ Objective To investigate mechanisms of Danggui Shaoyao Powder in treating premature ovarian insufficiency(POI). Methods The rat model of POI was constructed by intraperitoneal injection of cyclophosphamide(CTX). The blank group, model group, gonadotropin-releasing hormone analogue(GnRHa) group(with intraperitoneal injection of GnRHa at 0.1 mg/kg on the 10th and 5th days before constructing the model) and Danggui Shaoyao Powder group(with gavage administration of Danggui Shaoyao Powder at 10 mL/kg after constructing the model) were set up, with 6 rats in each group, and continuous intervention for 14 days. The morphological structure of rat ovarian tissues was observed by using hematoxylin-eosin(HE) staining and transmission electron microscopy. The levels of serum follicle-stimulating hormone(FSH), estradiol(E2), and anti-Müllerian hormone(AMH) were detected by using enzyme-linked immunosorbent assay(ELISA). The expression levels of key factors in the transforming growth factor β1(TGFβ1)-Smad2 signaling pathway in ovarian tissues were detected by using Western blot assay and reverse transcription-polymerase chain reaction(RT-PCR) assay. Results In the rats of the model group, the levels of serum E2 and AMH were decreased significantly, while the level of FSH was increased significantly, with an increase in atretic follicles, granulosa cells aligned loosely, severe edema of organelles, disordered tissue spaces, cell membrane damage, and irregular depression of cell nuclei in ovarian tissues, indicating that the ovarian function was severely impaired. After the interventions with Danggui Shaoyao Powder and GnRHa, the levels of serum E2 and AMH in the POI model rats were rebounded, while the level of FSH was decreased, with reduced vacuoles in ovarian tissues, increased ovarian follicles at all levels, damages to cell membrane and nuclear membrane similar to those in the model group, and alleviated swelling of organelles. The expression levels of phosphorylated transforming growth factor β receptor 1(p-TGFβR1) and phosphorylated Smad2(p-Smad2) protein in the ovarian tissues of the model group were significantly increased(P<0.05), and the expression levels of TGFβ1, transforming growth factor β receptor 1(TGFβR1), transforming growth factor β receptor 2(TGFβR2), Smad2 protein and mRNA were significantly increased(P<0.05), and the expression levels of Smad6 and Smad7 proteins and mRNA were significantly decreased(P<0.05), indicating that the TGFβ1-Smad2 signaling pathway was overactivated. After the intervention with Danggui Shaoyao Powder, the expression levels of Smad2 and p-Smad2 proteins were significantly decreased(P<0.05), and the expression levels of TGFβ1, TGFβR1, TGFβR2 proteins and mRNA were significantly decreased(P<0.05). The expression levels of Smad6 and Smad7 proteins and mRNA were significantly increased(P<0.05). The indicator changes after the intervention of Danggui Shaoyao Powder were similar to those after the intervention of GnRHa. Conclusion Danggui Shaoyao Powder may exert its therapeutic effect on POI by inhibiting the abnormal activation of the TGFβ1-Smad2 signaling pathway. |
| Key words: Danggui Shaoyao Powder Premature ovarian insufficiency(POI) Transforming growth factor β1(TGFβ1)-Smad2 signaling pathway Animal experiment |
