中药药理与临床

目的:利用网络毒理学筛选雷公藤致肾毒性靶点，利用网络药理学与分子对接探讨黄芪减雷公藤肾毒性的作用机制，并进行动物实验验证。方法:在OMIM、PharmGKB、Gene Cards收集肾毒性相关靶点，在CTD检索雷公藤肾毒性靶点，Venny2.1.0获得雷公藤肾毒性交集靶点;检索TCMSP、ETCM、以及文献补充收集黄芪成分，Swiss Target Prediction收集黄芪成分作用靶点，Venny2.1.0获得黄芪干预雷公藤肾毒性交集靶点;通过Cytoscape3.9.1构建网络图;利用Metascape对交集靶点进行GO功能和KEGG通路富集; Autodock4.2进行分子对接。根据预测结果进行动物实验验证。结果:收集黄芪化学成分46个、雷公藤致肾毒性靶点425个、黄芪干预雷公藤肾毒性潜在靶点83个。富集分析发现共同靶点主要参与细胞对化学应激的反应、细胞迁移、细胞运动等生物工程。KEGG信号通路排除广泛通路后主要富集于AGE-RAGE、HIF-1、C型凝集素受体、IL-17、PI3K/AKT等信号通路。PPI结果显示AKT1、IL6、VEGFA可能是黄芪干预雷公藤肾毒性的核心靶点，其中AKT1为最重要的核心靶点。“中药-成分-靶点-通路”结果显示异鼠李素、华良姜素、山柰酚、槲皮素、3-羟基-9,10-二甲氧基紫檀烷可能是黄芪干预雷公藤肾毒性的核心成分。分子对接结果发现黄芪中以上核心成分与核心靶点结合后，均与AKT1结合最强。动物实验结果表明模型对照组大鼠出现急性肾损伤病理表现，血清SCr、BUN含量显著升高(P<0.01)，雷公藤水提物能够明显下调Pi3k、Akt mRNA表达，而配伍黄芪水提物后能够上调其表达。结论:黄芪中核心成分可能作用于AKT1核心靶点，通过调控PI3K/AKT通路发挥干预雷公藤肾毒性的作用。

Objective: Network toxicology was used to screen the nephrotoxic targets of Tripterygium wilfordii. Network pharmacology and molecular docking were used to explore the mechanism of ASTRAGALI RADIX in reducing the nephrotoxicity of Tripterygium wilfordii,and an animal experiment was performed for verification. Methods: The nephrotoxicity-related targets were collected in OMIM,PharmGKB,and Gene Cards,and the nephrotoxic targets of Tripterygium wilfordii were searched in the CTD database. Venny2.1.0 was used to take the intersection of nephrotoxic targets of Tripterygium wilfordii. TCMSP,ETCM,and literature were searched to collect the chemical components of ASTRAGALI RADIX. The action targets of the chemical components of ASTRAGALI RADIX were collected by Swiss Target Prediction. The intersection targets of ASTRAGALI RADIX interfering with the nephrotoxicity of Tripterygium wilfordii were obtained by Venny2.1.0. The network map was constructed by Cytoscape 3.9.1. GO function and KEGG pathway were enriched by Metascape for the intersection targets. Autodock 4.2 was used for molecular docking. According to the predicted results,the experimental verification was carried out,and the serum was detected. In addition,the pathological changes were observed,and the path in KEGG analysis was verified. Results: A total of 46 chemical components of ASTRAGALI RADIX,425 nephrotoxic targets of Tripterygium wilfordii,and 83 potential targets of ASTRAGALI RADIX interfering with Tripterygium wilfordii nephrotoxicity were collected. The enrichment analysis suggested that the common targets were involved in bioengineering,such as the response of cells to chemical stress,cell migration,and cell movement. KEGG signaling pathway was mainly enriched in AGE-RAGE,HIF-1,C-type lectin receptor,IL-17,PI3K/AKT,and other signaling pathways after exclusion of extensive pathways. The PPI results showed that AKT1,IL6,and VEGFA might be the core targets of ASTRAGALI RADIX interfering with Tripterygium wilfordii nephrotoxicity,in which AKT1 was the most important core target. The results of traditional Chinese medicine-component-target-pathway showed that isorhamnetin,huperzine,kaempferol,quercetin,3-hydroxy-9,and 10-dimethoxypyrantel might be the core components of ASTRAGALI RADIX in interfering with the nephrotoxicity of Tripterygium wilfordii. The molecular docking results showed that after the core targets were bound with core components of ASTRAGALI RADIX,they all had good binding activity with AKT1. The results of animal experiments indicated that acute renal injury occurred in the model control group. The serum contents of SCr and BUN were significantly increased( P<0.01). Tripterygium wilfordii water extract significantly down-regulated the mRNA expressions of PI3K and AKT,while combined with ASTRAGALI RADIX water extract,the mRNA expressions of PI3K and AKT were up-regulated. Conclusion: Core components of ASTRAGALI RADIX may act on the core targets of AKT1,and they may interfere with the nephrotoxicity of Tripterygium wilfordii by regulating the PI3K/AKT pathway.