基于网络药理学研究归芪补血口服液抗HIV-1作用及机制  

Study on the anti-HIV-1 effect and mechanism of Guiqibuxue oral liquid based on network pharmacology

作　　者：覃秋珍 詹妤婕 王倩 刘洁 刘欣 周波 王捷 叶力 Qin Qiuzhen;Zhan Yujie;Wang Qian;Liu Jie;Liu Xin;Zhou Bo;Wang Jie;Ye Li(Guangxi Key Laboratory of AIDS Prevention and Treatment,Life Science Institute,Guangxi Medical University,Nanning 530021,China;School of Public Health,Guangxi Medical University,Nanning 530021,China;Pharmacy Factory of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学生命科学研究院,广西艾滋病防治研究重点实验室,南宁530021 [2]广西医科大学公共卫生学院,南宁530021 [3]广西医科大学制药厂,南宁530021

出　　处：《广西医科大学学报》2022年第12期1891-1902,共12页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81860655);广西重点研发项目资助(No.桂科AB19245038);广西高校中青年教师科研基础能力提升项目(No.2022KY0090)。

摘　　要：目的:通过体外实验研究归芪补血口服液复方制剂的抗HIV-1活性,并基于网络药理学挖掘潜在的作用机制,为开展中药制剂的药理研究提供理论依据。方法:通过ATP法评估药物对TZM-bl细胞和MT-2细胞的毒性作用;利用TZM-bl-HIV-1IIIB、MT-2-HIV-1IIIB两种细胞感染模型,通过荧光素酶活性检测试剂检测病毒活性从而评估药物抗HIV-1活性。基于网络药理学预测关键靶点和关键通路,利用分子对接模拟技术探索关键成分与关键靶点的相互作用,并通过实时荧光定量PCR(RT-qPCR)检测相关通路的关键靶点mRNA相对表达量。结果:归芪补血口服液对TZM-bl和MT-2两种细胞的50%细胞毒性浓度(CC50)分别是(13588±2633)μg/mL、(9846±2526)μg/mL;50%病毒抑制浓度(IC50)分别是(690.50±118.00)μg/mL、(585.80±176.00)μg/mL;SI(CC50/IC50)分别为19.60±1.70、17.03±0.86。网络药理学筛选出归芪补血口服液主要活性成分有槲皮素、山奈酚、木犀草素、豆甾醇、beta-谷甾醇等,关键靶点包括IL-6、IL1B、AKT1、JUN、HSP90AA1等,靶点富集的通路包括NF-κB信号通路、Toll-like受体信号通路和NOD-like受体信号通路等免疫及炎症相关的通路。分子对接结果显示,归芪补血口服液主要有效成分和靶点具有较稳定的结合活性;RT-qPCR实验结果显示,归芪补血口服液可下调IKBKB、FOS、TLR3、TLR8的mRNA表达,上调NFKBIA的mRNA表达。结论:归芪补血口服液复方制剂具有一定的抗HIV-1活性,可能通过调控IKBKB、NFKBIA、TLR3、TLR8和FOS等潜在靶点的mRNA表达水平来发挥抗HIV-1作用,其作用机制可能与NF-κB等信号通路相关。Objective:To investigate the anti-HIV-1 activity of the traditional Chinese herbs Guiqibuxue oral liquid in vitro,and explore the potential mechanism based on network pharmacology,so as to provide a reference for the pharmacological research of traditional Chinese medicine.Methods:The cytotoxicity of Guiqibuxue oral liquid on TZM-bl and MT-2 cells was evaluated by ATP assay.TZM-bl-HIV-1IIIB and MT-2-HIV-1IIIB cell infection models were established to evaluate the anti-HIV-1 activity by detecting the expression of luciferase activity.The key targets and key pathways were predicted based on network pharmacology,the interaction between key components and key targets was explored using molecular docking,and the relative mRNA expressions were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results:The concentration of cytotoxicity 50%(CC50)of Guiqibuxue oral liquid to TZM-bl and MT-2 cells were(13,588±2,633)μg/mL and(9,846±2,526)μg/mL,respectively.The half maximal inhibitory concentration(IC50)of the two HIV-cell models were(690.50±118.00)μg/mL and(585.80±176.00)μg/mL,respectively.The SI(CC50/IC50)was 19.60±1.70 and 17.03±0.86,respectively.The main active ingredients of Guiqibuxue oral liquid were screened by network pharmacology,including quercetin,kaempferol,luteolin,stigmasterol,beta-sitosterol,etc.The key targets included IL-6,IL1B,AKT1,JUN,HSP90AA1,etc.The target genes were enriched in the pathways including NF-κB signaling pathway,Toll-like receptor signaling pathway,NOD-like receptor signaling pathway and other immune and inflammatory-related pathways.The molecular docking results showed that the main active components and targets of Guiqibuxue ora l liquid had stable binding activity.The results of RT-qPCR experiment showed that the mRNA relative expressions of IKBKB、FOS、TLR3、TLR8 were down-regulated by Guiqibuxue oral liquid,and the mRNA expression of NFKBIA was up-regulated.Conclusion:Guiqibuxue oral liquid has certain anti-HIV-1 activity,which mainly inhib

关 键 词：归芪补血口服液 HIV 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学]