基于网络药理学探究合欢皮-白蒺藜药对抑制肝星状细胞系LX2的抗焦亡作用  

Anti-pyroptosis effect of Albiziae Cortex-Tribuli Fructus combination on hepatic stellate cell line LX2:based on network pharmacology

作　　者：谢泽宇 许一笑 郑梦圆 郑静茹 姚立[1] XIE Ze-yu;XU Yi-xiao;ZHENG Meng-yuan;ZHENG Jing-ru;YAO Li(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053

出　　处：《中国中药杂志》2023年第2期481-491,共11页China Journal of Chinese Materia Medica

基　　金：浙江省自然科学基金项目(LY21H280003)。

摘　　要：基于网络药理学、分子对接技术和体外实验验证探讨合欢皮-白蒺藜药对对肝星状细胞系(HSC)-LX2焦亡及肝纤维化的影响。在网络药理学中,通过在线数据库和中国知网获取合欢皮、白蒺藜以及肝纤维化的相关靶点,构建“药物-成分-靶点”网络和“药物-成分-靶点-疾病”网络,使用STRING数据库构建蛋白互作网络,采用Metascape数据库进行GO和KEGG数据分析,预测合欢皮-白蒺藜药对治疗肝纤维化的作用机制,并采用分子对接技术对网络药理学部分结果进行初步验证,再进一步结合体外实验验证上述结论并深入研究。从网络药理学分析结果可知,合欢皮-白蒺藜药对有25个有效成分和439个靶点,与肝纤维化共同靶点152个,涉及NLRP3和caspase-1等。KEGG通路富集分析得到194条信号通路,其中NOD样受体信号通路位于前列,结合共同靶点与细胞焦亡有关。体外实验结果显示,合欢皮-白蒺藜药对的含药血清能够降低HSC-LX2的增殖率以及ROS、IL-18和IL-1β含量(P<0.05)。Western blot和qRT-PCR结果显示,血管紧张素Ⅱ(AngⅡ)刺激后HSC-LX2中NLRP3、caspase-1、α-SMA及GSDMD蛋白和基因表达升高,合欢皮-白蒺藜药对的含药血清干预后表达下降(P<0.05)。综上,合欢皮-白蒺藜药对的含药血清可抑制细胞焦亡的经典通路,该机制可能参与其抗肝纤维化作用,具体机制有待后续进一步研究。这与网络药理学预测的结果相符。Based on network pharmacology,molecular docking,and in vitro experimental verification,this study aims to explore the effect of Albiziae Cortex-Tribuli Fructus combination on HSC-LX2 pyroptosis.Specifically,the targets of Albiziae Cortex,Tribuli Fructus,and hepatic fibrosis were retrieved from an online database and CNKI,and"drug-component-target"network and"drug-component-target-disease"network were constructed.Protein-protein interaction(PPI)network was established based on STRING.Metascape was employed for Gene Ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment,and the mechanism of Albiziae Cortex-Tribuli Fructus combination against liver fibrosis was predicted.Molecular docking was used to verify some of the results of network pharmacology,and in vitro experiment was carried out to further verify the above conclusions.According to the results of network pharmacological analysis,25 active components and 439 targets of Albiziae Cortex-Tribuli Fructus combination and 152 anti-liver fibrosis targets were screened out,including nucleotide-binding oligomerization domain and leucine-rich-repeat-and pyrin-domain-containing 3(NLRP3)and caspase-1.The key targets were involved in 194 KEGG pathways in which the NOD-like receptor signaling pathway topped.The binding common targets were related to pyroptosis.The results of in vitro experiment showed that the pair-containing serum reduced the proliferation rate of HSC-LX2 and the content of reactive oxygen species(ROS),interleukin-18(IL-18),and interleukin-1β(IL-1β)(P<0.05).Western blot and qRT-PCR suggested that the protein and gene expression of NLRP3,caspase-1,α-smooth muscle actin(α-SMA),and gasdermin D(GSDMD)in HSC-LX2 increased after AngⅡstimulation,and the expression decreased after the intervention of pair-containing serum(P<0.05).In summary,the pair-containing serum can inhibit the classic pathway of pyroptosis,which may be the anti-liver fibrosis mechanism.This is consistent with the predicted results of network p

关 键 词：合欢皮-白蒺藜 网络药理学 细胞焦亡 肝纤维化 HSC-LX2 

分 类 号：R285[医药卫生—中药学]