Yan Zhi-Yi, Zong Yong-Hua, Zhang Cheng-Fei, Wu Li-Li, Qin Ling-Ling, Liu Tong-Hua
Dongfang Hospital, Beijing University of Chinese Medicine Beijing 100078, China Key Laboratory of Health-cultivation by Ministry of Education of the People's Republic of China,Beijing University of Chinese Medicine Beijing 100029, China.
Department of Tibetan Medicine, University of Tibetan Medicine Lhasa 850000, China Key Laboratory of Health-cultivation by Ministry of Education of the People's Republic of China,Beijing University of Chinese Medicine Beijing 100029, China.
Zhongguo Zhong Yao Za Zhi. 2023 May;48(9):2538-2551. doi: 10.19540/j.cnki.cjcmm.20230203.501.
To explore the mechanism of the active ingredients of Qishiwei Zhenzhu Pills in inhibiting the hepatorenal toxicity of the zogta component based on serum pharmacochemistry and network pharmacology, thereby providing references for the clinical safety application of Qishiwei Zhenzhu Pills. The small molecular compounds in the serum containing Qishiwei Zhenzhu Pills of mice were identified by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Then, by comprehensively using Traditional Chinese Medicines Systems Pharmacology(TCMSP), High-throughput Experiment-and Reference-guided Database(HERB), PubChem, GeneCards, SuperPred, and other databases, the active compounds in the serum containing Qishiwei Zhenzhu Pills were retrieved and their action targets were predicted. The predicted targets were compared with the targets of liver and kidney injury related to mercury toxicity retrieved from the database, and the action targets of Qishiwei Zhenzhu Pills to inhibit the potential mercury toxicity of zogta were screened out. Cytoscape was used to construct the active ingredient in Qishiwei Zhenzhu Pills-containing serum-action target network, and STRING database was used to construct the protein-protein interaction(PPI) network of intersection targets. The Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out on the target genes by the DAVID database. The active ingredient-target-pathway network was constructed, and the key ingredients and targets were screened out for molecular docking verification. The results showed that 44 active compounds were identified from the serum containing Qishiwei Zhenzhu Pills, including 13 possible prototype drug ingredients, and 70 potential targets for mercury toxicity in liver and kidney were identified. Through PPI network topology analysis, 12 key target genes(HSP90AA1, MAPK3, STAT3, EGFR, MAPK1, APP, MMP9, NOS3, PRKCA, TLR4, PTGS2, and PARP1) and 6 subnetworks were obtained. Through GO and KEGG analysis of 4 subnetworks containing key target genes, the interaction network diagram of active ingredient-action target-key pathway was constructed and verified by molecular docking. It was found that taurodeoxycholic acid, N-acetyl-L-leucine, D-pantothenic acid hemicalcium, and other active ingredients may regulate biological functions and pathways related to metabolism, immunity, inflammation, and oxidative stress by acting on major targets such as MAPK1, STAT3, and TLR4, so as to inhibit the potential mercury toxicity of zogta in Qishiwei Zhenzhu Pills. In conclusion, the active ingredients of Qishiwei Zhenzhu Pills may have a certain detoxification effect, thus inhibiting the potential mercury toxicity of zogta and playing a role of reducing toxicity and enhancing effect.
基于血清药物化学和网络药理学探讨七十味珍珠丸活性成分抑制佐塔成分肝肾毒性的作用机制，为七十味珍珠丸临床安全应用提供参考。采用高效液相色谱-串联质谱法（HPLC-MS/MS）鉴定含七十味珍珠丸小鼠血清中的小分子化合物。然后，综合运用中药系统药理学数据库（TCMSP）、高通量实验与参考指导数据库（HERB）、PubChem、GeneCards、SuperPred等数据库，检索含七十味珍珠丸血清中的活性成分并预测其作用靶点。将预测靶点与从数据库中检索到的汞毒性相关肝肾损伤靶点进行比较，筛选出七十味珍珠丸抑制佐塔潜在汞毒性的作用靶点。利用Cytoscape构建含七十味珍珠丸血清活性成分-作用靶点网络，利用STRING数据库构建交集靶点的蛋白质-蛋白质相互作用（PPI）网络。通过DAVID数据库对靶点基因进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析。构建活性成分-靶点-通路网络，筛选关键成分和靶点进行分子对接验证。结果显示，从含七十味珍珠丸血清中鉴定出44种活性化合物，其中包括13种可能的原型药物成分，鉴定出70个肝肾汞毒性潜在靶点。通过PPI网络拓扑分析，得到12个关键靶基因（HSP90AA1、MAPK3、STAT3、EGFR、MAPK1、APP、MMP9、NOS3、PRKCA、TLR4、PTGS2和PARP1）和6个子网络。通过对包含关键靶基因的4个子网络进行GO和KEGG分析，构建活性成分-作用靶点-关键通路相互作用网络图并进行分子对接验证。发现牛磺脱氧胆酸、N-乙酰-L-亮氨酸、D-泛酸半钙等活性成分可能通过作用于MAPK1、STAT3、TLR4等主要靶点，调节与代谢、免疫、炎症和氧化应激相关的生物学功能和通路，从而抑制七十味珍珠丸中佐塔的潜在汞毒性。综上所述，七十味珍珠丸活性成分可能具有一定解毒作用，从而抑制佐塔潜在汞毒性，发挥减毒增效作用。
