Hu Luanqian, Chen Yuqi, Chen Tingting, Huang Dan, Li Shihua, Cui Shuna
Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Medical College of Yangzhou University, Yangzhou, China.
Department of Gynecology and Obstetrics, Affiliated Hospital of Yangzhou University, Yangzhou, China.
Front Pharmacol. 2020 Dec 4;11:582520. doi: 10.3389/fphar.2020.582520. eCollection 2020.
To investigate the mechanism of (Oliv.) Rehder & E.H.Wilson () and () against Pelvic Inflammatory Disease with Dampness-Heat Stasis Syndrome via network pharmacological approach and experimental validation. The active compounds with OB ≥ 30% and DL ≥ 0.18 were obtained from TCMSP database and further confirmed by literature research. The targets of the compounds and disease were acquired from multiple databases, such as GeneCards, CTD and TCMSP database. The intersection targets were identified by Venny software. Cytoscape 3.7.0 was employed to construct the protein-protein interaction (PPI) network and compound-target network. Moreover, GO enrichment and KEGG pathway analysis were analyzed by DAVID database. Finally, CCK-8, Griess assay and a cytometric bead array (CBA) immunoassay were used for experimental validation by detecting the influence of the active compounds on proliferation of macrophage, release of NO and TNF-α after LPS treatment. 9 bioactive compounds were identified from and . Those compounds corresponded to 134 targets of pelvic inflammatory disease with dampness-heat stasis syndrome. The targets include vascular endothelial growth factor A (VEGFA), von willebrand factor (VWF), interleukin 6 (IL6), tumor necrosis factor (TNF) and nuclear transcription factor 1 (NFκB1). They act on the signaling pathways like advanced glycation end products-receptor of advanced glycation end products (AGE-RAGE), focal adhesion (FA), Toll-like receptor (TLR) and nuclear transcription factor κB (NF-κB). In addition, by validation, the selected active components of and such as acacetin, kaempferol, linarin, isovitexin, sinoacutine could significantly inhibit the release of NO induced by LPS, respectively. Moreover, different dose of acacetin, kaempferol, isovitexin and sinoacutine significantly inhibits the TNF-α production. This study provides solid evidence for the anti-inflammatory mechanism of and against pelvic inflammatory disease with dampness-heat stasis syndrome, which will provide a preliminary evidence and novelty ideas for future research on the two herbs.
通过网络药理学方法和实验验证，研究（Oliv.）Rehder & E.H.Wilson（）和（）对湿热瘀结型盆腔炎的作用机制。从中药系统药理学数据库（TCMSP）中获取口服生物利用度（OB）≥30%且药物相似性（DL）≥0.18的活性成分，并通过文献研究进一步确认。化合物和疾病的靶点从多个数据库获取，如GeneCards、CTD和TCMSP数据库。利用Venny软件确定交集靶点。采用Cytoscape 3.7.0构建蛋白质-蛋白质相互作用（PPI）网络和化合物-靶点网络。此外，通过DAVID数据库进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）通路分析。最后，使用细胞计数试剂盒-8（CCK-8）、Griess法和细胞因子微球阵列（CBA）免疫分析法进行实验验证，检测活性成分对脂多糖（LPS）处理后巨噬细胞增殖、一氧化氮（NO）释放和肿瘤坏死因子-α（TNF-α）的影响。从和中鉴定出9种生物活性成分。这些化合物对应于湿热瘀结型盆腔炎的134个靶点。靶点包括血管内皮生长因子A（VEGFA）、血管性血友病因子（VWF）、白细胞介素6（IL6）、肿瘤坏死因子（TNF）和核转录因子1（NFκB1）。它们作用于晚期糖基化终产物-晚期糖基化终产物受体（AGE-RAGE）、黏着斑（FA）、Toll样受体（TLR）和核转录因子κB（NF-κB）等信号通路。此外，通过验证，所选的和的活性成分如刺槐素、山柰酚、蒙花苷、异荭草素、华蟾酥毒基可分别显著抑制LPS诱导的NO释放。此外，不同剂量的刺槐素、山柰酚、异荭草素和华蟾酥毒基可显著抑制TNF-α的产生。本研究为和对湿热瘀结型盆腔炎的抗炎机制提供了确凿证据，为这两种草药的未来研究提供了初步证据和新颖思路。
