目的 采用UPLC-Q-TOF-MS/MS技术结合网络药理学及体外细胞实验，探讨九制黄精与生黄精对脂多糖（LPS）诱导的RAW 264.7巨噬细胞炎症反应的抑制作用差异。方法 通过UPLC-Q-TOF-MS/MS技术结合文献检索，对比分析九制黄精与生黄精的化学成分差异；基于已鉴定成分筛选黄精抗炎的关键活性成分与核心作用靶点，构建“中药-成分-靶点-疾病”调控网络，并通过分子对接验证核心成分与靶点的结合能力；将不同质量浓度(1、10、50、100、200、400、800μg·mL^-1)的九制黄精、生黄精水提物（NPA、RPA）分别作用于RAW 264.7细胞24 h，采用CCK-8法检测细胞活力；使用1μg·mL^-1LPS诱导RAW 264.7细胞建立炎症模型，实验设对照组、模型组(1μg·mL^-1 LPS)、RPA组(100、200、400μg·mL^-1)及NPA组(100、200、400μg·mL^-1)，采用ELISA法检测各组细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）的分泌水平，比较二者抗炎作用差异。结果 共鉴定出114种化学成分，其中82种为九制黄精与生黄精共有，21种为九制黄精特有成分，11种为生黄精特有成分；网络药理学分析筛选获得873个黄精抗炎相关交集靶点，确定延龄草苷、N-阿魏酰真胺、麝香草酚、丁香脂素、静特诺皂苷元为核心活性成分，转导和转录激活因子（STAT3）、蛋白激酶B（AKT1）、缺氧诱导因子1亚基α（HIF1A）、肿瘤坏死因子（TNF）、白细胞介素-6（IL-6）和B细胞淋巴瘤/白血病-2基因（BCL2）为核心靶点，基因本体（GO）功能注释与京都基因与基因组百科全书（KEGG）通路富集分析显示，磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）、高级糖基化终末产物（AGE）-受体（RAGE）信号通路是黄精发挥抗炎作用的潜在关键通路；体外细胞毒性实验表明，各质量浓度的NPA及400μg·mL^-1以下的RPA对RAW 264.7细胞存活率均无显著影响（P>0.05），相同质量浓度下NPA组细胞存活率略高于RPA组，但差异无统计学意义（P>0.05）；抗炎活性评价显示，与对照组相比，模型组细胞上清液中TNF-α、IL-6水平显著升高（P<0.01）；与模型组相比，NPA、RPA各质量浓度组均能显著抑制LPS诱导的TNF-α、IL-6分泌（P<0.01）；在400μg·mL^-1浓度下，RPA组TNF-α水平低于NPA组，而各质量浓度下RPA组IL-6分泌量均高于NPA组。结论 九制黄精的化学成分较生黄精更丰富，二者均能有效缓解LPS诱导的RAW 264.7细胞炎症反应，但在调控炎症因子分泌方面存在差异；九制黄精的抗炎作用可能与其特有成分及调控PI3K-Akt、AGE-RAGE等信号通路相关。

Objective To explore the differences in the inhibitory effects of nine-processed Polygonatum cyrtonema and raw Polygonatum cyrtonema on lipopolysaccharide（LPS）-induced inflammatory responses in RAW 264.7 macrophages by using UPLC-Q-TOF-MS/MS technology combined with network pharmacology and in vitro cell experiments. Methods The chemical composition differences between nine-processed P. cyrtonema and raw P. cyrtonema were compared and analyzed through UPLC-Q-TOF-MS/MS technology and literature retrieval. Based on the identified components, the key active components and core targets of P. cyrtonema for anti-inflammation were screened, and a "Chinese medicine-component-target-disease" regulatory network was constructed. Molecular docking was used to verify the binding ability of the core components and targets. Different concentrations(1, 10, 50, 100, 200, 400, 800 μg·mL^-1) of nine-processed P. cyrtonema and raw P. cyrtonema water extracts（NPA, RPA） were applied to RAW 264.7 cells for 24 hours, and cell viability was detected by the CCK-8 method. RAW 264.7 cells were induced by 1 μg·mL^-1 LPS to establish an inflammatory model. The experiment was set up with a control group, a model group(1 μg·mL^-1 LPS), a RPA group(100, 200, 400 μg·mL^-1), and an NPA group(100, 200, 400 μg·mL^-1). The secretion levels of tumor necrosis factor-α（TNF-α） and interleukin-6（IL-6） in the cell supernatants of each group were detected by ELISA, and the differences in anti-inflammatory effects between the two were compared. Results A total of 114 chemical components were identified, among which 82 were common to both nine-processed P. cyrtonema and raw P. cyrtonema, 21 were unique to nine-processed P. cyrtonema, and 11 were unique to raw P. cyrtonema. Network pharmacology analysis screened out 873 intersection targets related to the anti-inflammatory effects of P. cyrtonema. The core active components were identified as diosgenin glucoside, N-feruloyltrueamine, thymol, syringaresinol, gentrogenin. The core targets were determined as signal transducer and activator of transcription 3（STAT3）, protein kinase B（AKT1）, hypoxia-inducible factor 1 subunit alpha（HIF1A）, tumor necrosis factor（TNF）, interleukin-6（IL-6）, and B-cell lymphoma/leukemia-2 gene（BCL2）. Gene Ontology（GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis showed that the phosphatidylinositol 3-kinase/protein kinase（PI3K-Akt） and advanced glycation end product-receptor（AGE-RAGE） signaling pathways were the potential key pathways for the anti-inflammatory effects of P. cyrtonema. In vitro cytotoxicity experiments showed that different concentrations of NPA and RPA at concentrations below 400 μg·mL^-1 had no significant effect on the survival rate of RAW 264.7 cells（P ＞ 0.05）. At the same concentration, the survival rate of the NPA group was slightly higher than that of the RPA group, but the difference was not statistically significant（P ＞ 0.05）. Anti-inflammatory activity evaluation showed that compared with the control group, the levels of TNF-α and IL-6 in the cell supernatants of the model group were significantly increased（P ＜ 0.01）. Compared with the model group, all concentrations of NPA and RPA groups could significantly inhibit the secretion of TNF-α and IL-6 induced by LPS（P ＜ 0.01）. At a concentration of 400 μg·mL^-1, the TNF-α level in the RPA group was lower than that in the NPA group, while the secretion of IL-6 in the RPA group was higher than that in the NPA group at all concentrations. Conclusion The chemical composition of processed P. cyrtonema is more abundant than that of raw P. cyrtonema. Both can effectively alleviate LPS-induced inflammatory responses in RAW 264.7 cells, but there are differences in the regulation of inflammatory factor secretion. The anti-inflammatory effect of nine-processed P. cyrtonema may be related to its unique components and the regulation of PI3K-Akt, AGE-RAGE, and other signaling pathways.

R285.5;Q332

国家自然科学基金资助项目(82160827); 江西中医药大学科技创新团队发展计划资助项目(CXTD-22004); 中药鉴定学3D-MR虚拟仿真实验教学技术改进资助项目(2024SJSZ04)