益气养阴祛瘀中药对干燥综合征模型NOD小鼠TLR/IFN/BAFF通路的作用研究

目的观察益气养阴祛瘀中药对干燥综合征(SS)模型NOD小鼠外周血单个核细胞TLR9、IFN-α、BAFFm RNA表达水平的调节作用,探讨其对SS的TLR-IFN-BAFF通路信号表达的影响及其作用机制。方法选择NOD小鼠20只及ICR正常小鼠10只,饲养于无特殊病菌的恒温环境。NOD小鼠随机分为模型组、中药组,各10只,ICR正常小鼠10只,作为对照组。于小鼠9周龄时开始每日定时灌胃益气养阴祛瘀中药1次,持续12周。于实验第12周末时所有小鼠股动脉取血后处死,分离外周血单个核细胞,采用RT-PCR法检测TLR9、IFN-α、BAFF m RNA表达水平。小鼠股动脉取血后处死,摘取小鼠颌下腺,用光镜下观察小鼠颌下腺病理改变。结果对照组、模型组、中药组TLR9 m RNA表达水平分别为(0.15±0.03)、(0.92±0.05)、(0.29±0.06);对照组、模型组、中药组IFN-αm RNA表达水平分别为(0.12±0.03)、(1.48±0.04)、(0.20±0.03);对照组、模型组、中药组BAFF m RNA表达水平分别为(0.23±0.06)、(1.28±0.04)、(0.38±0.06);与正常对照组相比,模型组TLR9、IFN-α、BAFFm RNA均明显升高(P<0.01或P<0.05),与模型组比较,中药组TLR9、IFN-α、BAFFm RNA均明显下降(P<0.05)。结论 NOD小鼠存在血清TLR9、IFN-α、BAFF m RNA水平过度表达。益气养阴祛瘀中药对NOD小鼠TLR/IFN/BAFF通路的过度表达具有抑制作用。

Study on the compatibility principle of Wutou Decoction based on network pharmacology

Objective To investigate the underlying drug enhancement mechanisms of the Chuanwu(Aconiti Radix)and Huangqi(Astragali Radix)combination and toxicity reduction of Chuan-wu combined with Gancao(Glycyrrhizae Radix et Rhizoma)in Wutou Decoction(乌头汤,WTD),and to elucidate the compatibility principle.Methods The active compounds and potential effective targets of the selected combinations were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Traditional Chinese Medicines Integrated Database(TCMID).The toxicity of Chuanwu(Aconiti Radix)was investigated by selecting all five toxic compounds from the literature and the TCMSP database,and obtaining their targets through SwissTargetPrediction.Targets related to rheumatoid arthritis(RA)were searched using Dis-GeNET,GenCards,and Online Mendelian Inheritance in Man(OMIM).Mutual targets between the drug pairs and RA were selected as potential RA therapy targets.The medicinally active compound-target network was constructed using Cytoscape 3.9.0.Gene ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrich-ment were performed using the Database for Annotation,Visualization,and Integrated Dis-covery(DAVID)platform.Results We obtained 191 active compound targets for Gancao(Glycyrrhizae Radix et Rhizoma),171 for Huangqi(Astragali Radix),and 103 for Chuanwu(Radix Aconiti)(hypo-aconitine’s target was obtained through literature and SwissTargetPrediction).A total of 5872 genes were obtained for RA.A drug-active compound-target network involving 13 effect-en-hancing and nine toxicity reduction targets was constructed.PGR was the main effect en-hancement target,and KCNH2 was the main toxicity reduction target.The effect-enhancing targets were related to 23 GO terms(such as positive regulation of transcription from RNA polymerase II promoter,steroid hormone-mediated signaling pathway,plasma membrane,and protein binding)(P<0.01),and 13 KEGG pathways related to synergism[such as estro-gen signaling pathway,cholinergic synapse,and phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway].The toxicity reduction targets were related to 28 GO terms(mainly involes G-protein coupled receptor signaling pathway,plasma membrane,and drug binding)(P<0.01),and five KEGG pathways related to toxicity reduction(cholinergic syn-apse,calcium signaling pathway,regulation of actin cytoskeleton,neuroactive ligand-recept-or interaction,and serotonergic synapse).Conclusion The combination of Chuanwu(Aconiti Radix)and Huangqi(Astragali Radix)plays an important effect-enhancing role in WTD and involves the estrogen and PI3K/Akt sig-naling pathways,with PGR as the core.The Chuanwu(Aconiti Radix)and Gancao(Gly-cyrrhizae Radix et Rhizoma)combination decreases toxicity in WTD and is associated with the cholinergic synapse and calcium signaling pathways,with KCNH2 as the core.