Department of urology, Jinhua Hospital of TCM, Jinhua 321000, China.
Zhejiang Chinese Medical University college of pharmacy, Hangzhou 310053, China.
J Tradit Chin Med. 2023 Jun;43(3):533-541. doi: 10.19852/j.cnki.jtcm.2023.03.003.
To explored the mechanism of Buzhong Yigi decoction ( BZYQD) in inhibiting prostatic cell proliferation effect.
The compounds of BZYQD consisted with eight herbs were searched in TCMSP databases and the putative targets of BZYQD were collected in Drugbank database. Then, "Benign prostatic hyperplasia" (BPH) was used to find the targets based on the GeneCards, Online Mendelian Inheritance in Man (OMIM) and Therapeutic Target Database (TTD) databases, and they were further used to collect further collect the intersection targets between BZYQD and BPH by counter-selection. Next, Herb-Compound-Target-Disease network was constructed by Cytoscape software and protein interaction network was built by Search tool for recurring instances of neighbouring genes (STRING) database. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were analyzed by Database for Annotation, Visualization and Integrated Discovery (DAVID) database to predict the mechanism of the intersection targets. Mitogen activated protein kinase 8 (MAPK8), interleukin 6 (IL-6) and quercetin were chosen to perform molecular docking. Then 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was to detect the bility of BPH-1 (BPH epithelial cell line) by treated with quercetin at the concentrations of 15, 30, 60, 120 μM for 12, 24, 48, 72 h. The production of IL-6, tumor necrosis factor-α (TNF-α), IL-1β and were mRNA expression detected by enzyme-linked immunosorbent assay kit and quantitative real-time polymerase chain reaction. Western blot was used to detect the expression of phospho-p38 mitogen-activated protein kinase (p-P38) and matrix metalloprotein-9 (MMP-9).
A total 151 chemical ingredients of 8 herbs and 1756 targets in BZYQD, 105 common targets of BZYQD and BPH which mainly involving with MAPK8, IL-6, and so on. GO enrichment analysis got 352 GO entries (0.05) which included 208 entries of biological process, 64 entries of cell component and 80 entries of molecular function. KEGG pathway Enrichment analyses got 20 significant pathways which mainly involved with MAPK signaling way. MTT assay indicated quercetin inhibited the viability of BPH-1 cells by time-and dose-dependent manner. Quercetin decreased the IL-6, TNF-α and IL-1β production and mRNA expression, and the expression of p-P38 and MMP-9 were also obviously reduced after treated with quercetin.
BZYQD inhibited BPH through suppressing inflammatory response which might involving with regulating the MAPK signaling way.
探讨补中益气汤抑制前列腺细胞增殖的作用机制。
在 TCMSP 数据库中搜索补中益气汤的 8 种草药组成的化合物，并在 DrugBank 数据库中收集补中益气汤的潜在靶点。然后，根据 GeneCards、在线 Mendelian 遗传数据库（OMIM）和治疗靶点数据库（TTD）数据库，使用“良性前列腺增生（BPH）”查找靶点，并进一步使用反向选择收集补中益气汤和 BPH 的交集靶点。接下来，通过 Cytoscape 软件构建草药-化合物-靶点-疾病网络，通过 Search tool for recurring instances of neighbouring genes（STRING）数据库构建蛋白质相互作用网络。通过数据库 for Annotation, Visualization and Integrated Discovery（DAVID）数据库进行基因本体（GO）富集和京都基因与基因组百科全书（KEGG）通路富集分析，预测交集靶点的机制。选择丝裂原活化蛋白激酶 8（MAPK8）、白细胞介素 6（IL-6）和槲皮素进行分子对接。然后，通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）测定法检测浓度为 15、30、60、120 μM 的槲皮素处理后 BPH-1（BPH 上皮细胞系）的增殖能力 12、24、48、72 h。酶联免疫吸附测定试剂盒和实时定量聚合酶链反应检测白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）、白细胞介素 1β（IL-1β）的 mRNA 表达。通过 Western blot 检测磷酸化 p38 丝裂原活化蛋白激酶（p-P38）和基质金属蛋白酶-9（MMP-9）的表达。
补中益气汤的 8 种草药中共得到 151 种化学成分，1756 种靶点，105 种补中益气汤与 BPH 的共同靶点，主要涉及 MAPK8、IL-6 等。GO 富集分析得到 352 个 GO 条目（0.05），其中包括 208 个生物过程条目、64 个细胞成分条目和 80 个分子功能条目。KEGG 通路富集分析得到 20 个显著通路，主要涉及 MAPK 信号通路。MTT 测定表明，槲皮素呈时间和剂量依赖性抑制 BPH-1 细胞的活力。槲皮素降低了 IL-6、TNF-α 和 IL-1β 的产生和 mRNA 表达，用槲皮素处理后，p-P38 和 MMP-9 的表达也明显降低。
补中益气汤通过抑制炎症反应抑制 BPH，可能涉及调节 MAPK 信号通路。
