目的：基于网络药理学及实验验证探讨枸杞子治疗干眼(DE)的作用机制。

方法：以“枸杞子”为关键词，通过采用TCMSP数据库与分析平台，搜索枸杞子的药物活性成分及作用靶点，以GeneCards和OMIM数据库搜索Dry eye(DE)相关的基因靶点，将枸杞子与DE的靶点基因导入Venn软件，可获得两者交集的靶点图，后将数据导入String数据库获得PPI蛋白与蛋白相互作用网络图，借助于Cytoscape3.7.2软件构建枸杞子活性成分-作用靶点-相关疾病的网络图； 再利用Bioconductor平台及R语言GO富集分析、KEGG富集分析； 通过实验验证干眼发病机制中的关键靶标。

结果：通过TCMSP数据库与分析平台筛选得到枸杞子的有效化学成分45种，活性成分对应的靶点基因174个，与DE共同的靶点基因131个，根据“药物-成分-疾病-靶点”网络拓扑图，枸杞子治疗DE的主要有效成分27个。分析PPI网络，根据度值较高，即枸杞子治疗DE的关键靶点主要包括AKT1、VEGFA、CASP3、IL1B、JUN、PTGS2、CXCL8等，根据GO富集分析获得枸杞子治疗DE的166种生物学功能与过程，KEGG富集分析显示涉及31条信号通路，此外，通过实验验证发现DE模型组的结膜组织中AKT1、IL-6、TNF-α及IL-17蛋白表达量显著升高。

结论：枸杞子治疗DE是多成分-多靶点-多途径的复杂过程，且枸杞子治疗DE主要通过抗炎及抑制细胞凋亡相关分子参与调控。

METHODS: Taking “fructus lycii” as key words, the active ingredients and target of fructus lycii were searched by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Gene targets related to dry eye(DE)were searched by GeneCards and OMIM databases. The target genes of fructus lycii and DE were imported into Venn software to obtain the intersection target map of them. After that, the data were imported into the String database to obtain the PPI protein-protein interaction network diagram. Using Cytoscape3.7.2 software, the PPI protein-protein interaction network diagram was constructed for active ingredients, target sites and related diseases of fructus lycii. The Bioconductor platform and R language were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis. And the key targets in the pathogenesis of DE were verified by experiments.

RESULTS: Through TCMSP, 45 types of effective chemical components of fructus lycii, 174 target genes corresponding to active components and 131 common target genes with DE were screenedout. In accordance with the network topology of “drug-composition-disease-target”, 27 main effective components of fructus lycii were found in the treatment of DE. The PPI network was analyzed according to the high degree value, which is the key targets of fructus lycii for DE treatment, mainly including AKT1, VEGFA, CASP3, IL1B, JUN, PTGS2, CXCL8, etc. According to GO enrichment analysis, 166 biological functions and processes of fructus lycii for DE treatment were obtained. KEGG enrichment analysis showed that 31 signaling pathways were involved. Additionally, experimental verification displayed that the protein expressions of AKT1, interleukin-6(IL-6), tumor necrosis factor(TNF-α)and IL-17 in conjunctiva tissue of the DE model group were significantly increased.

CONCLUSIONS: Through network pharmacology, this study confirmed that the treatment of DE by fructus lycii is a complex process involving multi-components, multi-targets and multi-pathways, and that the treatment of DE by fructus lycii is mainly regulated by anti-inflammatory and apoptosis-related molecules.