Mechanism of Dendrobium Officinale Against Inflammatory Aging Based on Network Pharmacology and Molecular Docking
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摘要:
目的 基于网络药理学、分子对接和动物实验探讨铁皮石斛抗炎性衰老的作用机制。 方法 通过数据库ETCM、Pubchem、Swiss TargetPrediction获得石斛靶点，从Rat Genome Database网站下载衰老大鼠基因，应用 STRING数据库构建PPI蛋白互作网络；利用DAVID数据库对石斛作用于老年大鼠的潜在靶点进行GO、KEGG通路分析；应用分子对接方法对石斛主要活性成分和关键作用靶点进行对接。老年大鼠灌胃石斛溶液10周后ELISA检测IL-6、IL-10蛋白表达。 结果 筛选到主要活性成分齿叶黄皮素、吴茱萸碱、吴茱萸次碱、紫花前胡，关键靶点NOS3、STAT3 、AKT1、GSK3B，主要信号通路PI3K-AKT、Calcium、EGFR酪氨酸激酶抑制剂耐药性、HIF-1、Apelin、VEGF，分子对接显示主要活性成分和关键靶点之间均能自发结合，且形成稳定的氢键。石斛中剂量组IL-6表达量较空白对照组（P < 0.01）、低剂量组（P < 0.01）和百令胶囊组（P < 0.05）降低，低剂量组IL-10表达量较空白组（P < 0.05）升高。 结论 铁皮石斛发挥抗炎性衰老的功能可能与降低老年SD大鼠血清中IL-6的含量，增加血清中IL-10的含量有关，作用机制与石斛中齿叶黄皮素、吴茱萸次碱、紫花前胡、吴茱萸碱等调控关键基因AKT1、STAT3、NOS3、GSK3B表达有关。 Abstract:Objective To explore the mechanism of Dendrobium officinale against inflammatory aging based on network pharmacology and molecular docking and experimental verification. Methods We used related databases to obtain the action targets of the main components of Dendrobium officinale and the genes of aging rats, then intersected them to obtain the final target. STRING database was used to build a protein-protein interaction（PPI） network. DAVID database was used to conduct enrichment analysis of Gene Ontology（GO） and Kyoto Encyclopedia of Gene and Genomes（KEGG）. AutoDock software was to perform molecular docking verification on the main active components and key targets. Then, the aging rats were gavaged with dendrobium polysaccharide solution for ten week, the expression levels of IL-6 and IL-10 proteins were detected to verify the predicted targets. Results Through database retrieval, 28 active components and 492 target genes were screened out. There were 64 intersections. Enrichment analysis yielded 283 GO functional enrichment entries and 71 KEGG pathway enrichment entries. The main pathway is the PI3K-Akt signaling pathway, Calcium signaling pathway, EGFR tyrosine kinase inhibitor resistance, HIF-1 signaling pathway, Apelin signaling pathway, VEGF signaling pathway, etc. The results of molecular docking showed that each target（AKT1, STAT3, NOS3, GSK3B） and active ingredient（Dentatin, Rutaecarpine, Nodakenetin, Evodaiamine） spontaneously combined and formed stable hydrogen bonds. Animal experiments showed that the content of IL-6 in the medium dose group significantly decreased compared to the blank group, low dose group, and Bailing capsule group, and the content of IL-10 in the low dose group increased compared to the blank group. Conclusions Dendrobium officinale has a delaying effect on the decline of the immune system in elderly rats. It is mainly related to the decrease of IL-6 content and the increase of IL-10 content in the serum. Its mechanism may be related to the expression of key genes AKT1, STAT3, NOS3, GSK3B regulated by components in Dendrobium officinale, such as Dentatin, Rutaecarpine, Nodakenetin, and Evodaiamine. -
Key words:
- Dendrobium officinale /
- Inflammatory aging /
- Network pharmacology /
- IL-6 /
- IL-10
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图 1 网络药理学分析结果
A：活性成分-靶点网络图；B：成分靶点与衰老大鼠基因韦恩图；C：PPI互作网络图。
Figure 1. Network pharmacological analysis results
图 2 GO分析和KEGG分析结果
A:细胞组成；B：分子功能；C：生物过程；D：KEGG分析。
Figure 2. GO analysis and KEGG analysis result
图 5 不同给药对IL-6、IL-10的影响
A：IL-6表达比较；B：IL-10表达比较。*P < 0.05，**P < 0.01。
Figure 5. The effect of different susstances on the expression of IL-6、IL-10
表 1 靶点网络中Degree排名前10的靶点
Table 1. The top 10 target of Degree in the component-target network
靶点名称 Degree BC（中介中心性） CC（接近中心性） AKT1 43 0.235981041 0.746835443 MAPK3 37 0.081326682 0.694117647 STAT3 36 0.077079997 0.686046512 NOS3 29 0.053439074 0.621052632 GSK3B 28 0.045054203 0.621052632 PTGS2 25 0.050639569 0.584158416 MTOR 30 0.047754481 0.627659574 MMP9 26 0.020865854 0.584158416 ICAM1 22 0.026581134 0.561904762 VCAM1 18 0.019630032 0.541284404 
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表 2 靶点网络中Degree排名前9的成分
Table 2. The top 9 component of Degree in the component-target network
成分 Degree 名称 Dentatin 115 齿叶黄皮素 3-O-Methylgigantol 112 3-O-甲基甘醇 2-Hydroxy-4，7-Dimethoxy-9，10-Dihydrophenanthrene 108 2-羟基-4，7-二甲氧基-9，10-二氢菲 Rutaecarpine 108 吴茱萸次碱 Nodakenetin 108 紫花前胡 3，4，8-Trimethoxyphenanthrene-2，
5-Diol103 3，4，8-三甲氧基菲-2，5-二醇 Moscatilin 102 杓唇石斛素 Evodiamine 100 吴茱萸碱 Nootkatone 71 圆柚酮
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表 3 分子对接结合能（单位：Kcal/mol）
Table 3. Binding energy of molecular docking
靶点名称 齿叶黄皮素 吴茱萸次碱 紫花前胡 吴茱萸碱 AKT1 −5.12 −5.22 −5.56 −5.97 STAT3 −5.01 −4.77 −4.94 −4.94 NOS3 −5.06 −4.42 −4.37 −4.85 GSK3B −5.13 −5.04 −5.46 −5.38
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