【目的】探讨归元饮 （重用生地黄） 治疗干燥综合征的作用机制。 【方法】利用网络药理学相关数据库平台获取归元饮中 每味药物的有效成分，并同干燥综合征进行靶标预测，通过韦恩图确定两者的共同靶点，绘制“药物-成分-靶点”网络。 再次运用相同靶点构建蛋白相互作用 （PPI） 网络，然后在David平台进行基因本体论 （GO） 功能和京都基因与基因组百科全书 （KEGG） 富集分析，获取归元饮治疗干燥综合征的信号通路。最后通过分子对接初步验证核心成分与核心靶蛋白之间的结合 能。结合文献与分子对接的结果，进行小鼠体内外实验验证。 【结果】筛选得到有关归元饮的药物活性成分318个，与疾病的 交集靶点148个。GO和KEGG富集分析分别获取到921个条目和159条通路。分子对接结果显示豆甾醇与天冬氨酸特异性半 胱氨酸蛋白酶3 （CASP3） 的结合活性最好，槲皮素、山奈酚、β-谷甾醇等其他活性成分与核心靶点具有较好的结合活性。实 验验证结果显示归元饮及其活性成分豆甾醇可下调干燥综合征小鼠颌下腺组织及脂多糖 （LPS） 刺激颌下腺细胞中CASP3、核 苷酸结合寡聚化结构域样受体蛋白3 （NLRP3） 、凋亡相关斑点样蛋白 （ASC） 、白细胞介素1β （IL-1β） 、天冬氨酸特异性半胱 氨酸蛋白酶1 （Caspase-1） 蛋白表达水平并减少干燥综合征小鼠外周血及颌下腺细胞上清液中白细胞介素6 （IL-6） 、白细胞介 素1β （IL-1β） 和肿瘤坏死因子α （TNF-α） 含量。 【结论】通过网络药理学与分子对接的结果得出归元饮能通过核心成分豆甾醇 作用于CASP3等靶点发挥抗干燥综合征的疗效。小鼠体内外实验证明归元饮及其活性成分豆甾醇可以减轻干燥综合征小鼠 患病组织及细胞炎症，其可能通过靶向CASP3从而抑制NLRP3，改善干燥综合征。

Objective To investigate the therapeutic mechanism of Guiyuan Decoction （GYD，with Rehmanniae Radix as the principal herb） in Sjögren’ s syndrome （SS） . Methods Active components of GYD were retrieved from network pharmacology databases，and potential targets for SS were predicted. Common targets were identified via Venn analysis，followed by construction of a “herb-compound-target” network. Protein-protein interaction （PPI） networks were generated，and Gene Ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed using the DAVID platform. Molecular docking was employed to evaluate binding affinities between core components and key target proteins. In vivo and in vitro experiments were conducted to validate predictions based on literature and docking results. Results A total of 318 bioactive components in GYD and 148 disease-related overlapping targets were identified. GO and KEGG analyses yielded 921 functional terms and 159 signaling pathways， respectively. Molecular docking revealed stigmasterol exhibited the strongest binding affinity with caspase-3 （CASP3） ，while quercetin，kaempferol，and β-sitosterol also showed robust interactions with core targets. Experimental validation demonstrated that both GYD and its active constituent stigmasterol significantly downregulated protein expression levels of CASP3，NLR family pyrin domain containing 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β），and Caspase-1 in submandibular gland tissues of Sjögren’ s syndrome mice and in lipopolysaccharide （LPS）-stimulated submandibular gland cells. Furthermore， they effectively reduced the concentrations of interleukin-6 （IL-6），interleukin-1β （IL-1β），and tumor necrosis factor- α （TNF- α） in peripheral blood and cell culture supernatants of Sjögren’s syndrome mice. Conclusion Integrated network pharmacology and molecular docking suggest that GYD exerts anti-SS effects primarily via stigmasterol-mediated modulation of CASP3. Experimental evidence confirms that GYD and stigmasterol attenuate inflammatory responses and may ameliorate SS by targeting CASP3 to suppress NLRP3 inflammasome activation.

R285.5

国家自然科学基金资助项目 （编号：82305128）