■探讨补肾活血方(BSHX)治疗多囊卵巢综合征(PCOS)不孕症的作用靶点和作用机制。为中药复方的开发和临床应用提供依据。
■使用网络药理学-分子对接技术预测和验证BSHX治疗PCOS的活性成分和靶标。在动物实验中,将大鼠随机分为4组(对照组,模型组,BSHX组,二甲双胍组,每组n=16),来曲唑联合高脂乳剂灌胃建立PCOS大鼠模型。体重,阴道涂片,并记录各组大鼠的胚胎数量。苏木素-伊红(HE)染色观察卵巢及子宫内膜组织的形态学变化,采用酶联免疫吸附试验(ELISA)检测血清炎症因子水平。转化生长因子-β(TGF-β)的表达水平,转化生长因子β激活激酶1(TAK1),核因子κB(NF-κB),Vimentin,通过蛋白质印迹(WB)测量E-钙黏着蛋白。
■从BSHX获得90种活性药物成分,涉及201个蛋白质靶标,其中160个是潜在的治疗靶点。BSHX的活性成分与肿瘤坏死因子-α(TNF-α)的结合能较低,TGF-β,TAK1和NF-κB蛋白受体(<-5.0kcal/mol)。BSHX显著降低PCOS大鼠血清TNF-α水平(p<0.01),有效地调节了发情周期,恢复了卵巢和子宫内膜的病理变化,提高了怀孕率,增加了胚胎的数量。WB结果显示BSHX可以下调子宫内膜组织中TGF-β和NF-κB的蛋白表达水平(p<0.05)。促进E-cadherin蛋白的表达水平(p<.001),干预子宫内膜上皮-间质转化(EMT)过程。
■TGF-β,TAK1,NF-κB,TNF-α是BSHX治疗PCOS不孕的重要靶点。BSHX改善PCOS的炎症状态,通过TGF-β/NF-κB通路干预子宫内膜EMT过程,恢复子宫内膜病理变化,进一步改善PCOS患者的妊娠结局。
UNASSIGNED: To investigate the target and mechanism of action of Bushen Huoxue Recipe (BSHX) for the treatment of infertility in polycystic ovary syndrome (PCOS), to provide a basis for the development and clinical application of herbal compounds.
UNASSIGNED: Prediction and validation of active ingredients and targets of BSHX for the treatment of PCOS by using network pharmacology-molecular docking technology. In an animal experiment, the rats were randomly divided into four groups (control group, model group, BSHX group, metformin group, n = 16 in each group), and letrozole combined with high-fat emulsion gavage was used to establish a PCOS rat model. Body weight, vaginal smears, and number of embryos were recorded for each group of rats. Hematoxylin-eosin (HE) staining was used to observe the morphological changes of ovarian and endometrial tissues, and an enzyme-linked immunosorbent assay (ELISA) was used to detect the serum inflammatory factor levels. Expression levels of transforming growth factor-β (TGF-β), transforming growth factor beta activated kinase 1 (TAK1), nuclear factor kappa-B (NF-κB), Vimentin, and E-cadherin proteins were measured by western blot (WB).
UNASSIGNED: Ninety active pharmaceutical ingredients were obtained from BSHX, involving 201 protein targets, of which 160 were potential therapeutic targets. The active ingredients of BSHX exhibited lower binding energy with tumor necrosis factor-α (TNF-α), TGF-β, TAK1, and NF-κB protein receptors (< -5.0 kcal/mol). BSHX significantly reduced serum TNF-α levels in PCOS rats (p < .01), effectively regulated the estrous cycle, restored the pathological changes in the ovary and endometrium, improved the pregnancy rate, and increased the number of embryos. The results of WB suggested that BSHX can down-regulate protein expression levels of TGF-β and NF-κB in endometrial tissue (p < .05), promote the expression level of E-cadherin protein (p < .001), intervene in the endometrial epithelial-mesenchymal transition (EMT) process.
UNASSIGNED: TGF-β, TAK1, NF-κB, and TNF-α are important targets of BSHX for treating infertility in PCOS. BSHX improves the inflammatory state of PCOS, intervenes in the endometrial EMT process through the TGF-β/NF-κB pathway, and restores endometrial pathological changes, further improving the pregnancy outcome in PCOS.