We aimed to investigate therapeutic effect of Bushenhuoxue recipe in intrauterine adhesions (IUA) and explore the underlying molecular mechanism via integrating network pharmacology and in vitro experimental verification. The active compounds and gene targets of Bushenhuoxue recipe were screened in the TCMSP database and the IUA-related genes were identified using GeneCards database by the keyword “Intrauterine adhesions”. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted to reveal the underlying molecular mechanism of Bushenhuoxue recipe treating IUA. T-HESC cells were inducted to fibrotic state using TGF-β1 of 10 ng/ml concentration treating for 24 h. RT-qPCR or western blot was used to demonstrate the expression levels of fibrosis markers (COL1A1 and α-SMA) and KEGG pathway markers. Cell counting kit-8 (CCK8) assay was performed to illustrate the cell viability of endometrial stromal cell. The treatment of Bushenhuoxue recipe could significantly inhibit the proliferation and fibrosis of endometrial stromal cells. We obtained a total of 169 no-repeat ingredients of Bushenhuoxue recipe and 3044 corresponding targets. After taking intersection with 4230 no-repeat IUA-related genes, a total of 83 target genes related to both Bushenhuoxue recipe and IUA were finally identified. KEGG analysis found that PI3K-AKT signaling pathway might be the key pathway. Further experiment revealed that PI3K-AKT signaling pathway was significantly activated in endometrial stromal cells of fibrotic state and the treatment of Bushenhuoxue recipe could inhibit the PI3K-AKT signaling pathway. Further rescue assay demonstrated that Bushenhuoxue recipe suppressed the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway. Bushenhuoxue recipe suppresses the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway, eventually inhibiting the progression of IUA.

我们的目的是通过整合网络药理学和体外实验验证，探讨补肾活血方对宫腔粘连（IUA）的治疗作用，并探讨其潜在的分子机制。在TCMSP数据库中筛选补肾活血方的活性成分和基因靶点，并使用GeneCards数据库以“宫腔粘连”为关键词识别IUA相关基因。通过基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析，揭示补肾活血方治疗IUA的潜在分子机制。使用10ng/ml浓度的TGF-β1处理24小时，将T-HESC细胞诱导至纤维化状态。RT-qPCR或蛋白质印迹用于证明纤维化标志物（COL1A1和α-SMA）和KEGG通路标志物的表达水平。进行细胞计数试剂盒8（CCK8）测定以说明子宫内膜基质细胞的细胞活力。补肾活血方治疗可显着抑制子宫内膜基质细胞的增殖和纤维化。共获得补肾活血方的无重复成分169个及对应指标3044个。通过与4230个无重复IUA相关基因进行交叉分析，最终鉴定出83个与补肾活血方和IUA相关的靶基因。KEGG分析发现PI3K-AKT信号通路可能是关键通路。进一步实验发现，纤维化状态的子宫内膜基质细胞中PI3K-AKT信号通路显着激活，补肾活血方治疗可抑制PI3K-AKT信号通路。进一步的挽救实验表明，补肾活血方通过PI3K-AKT信号通路抑制子宫内膜基质细胞的增殖和纤维化。补肾活血方通过PI3K-AKT信号通路抑制子宫内膜基质细胞的增殖和纤维化，最终抑制IUA的进展。