目的 基于代谢组学探讨黄芪赤风汤对内皮细胞损伤的保护作用及其机制。方法 通过氧化低密度脂蛋白（ox-LDL）诱导小鼠脑微血管内皮bEnd.3细胞建立体外内皮细胞损伤模型。bEnd.3细胞随机分为对照组、模型组和黄芪赤风汤低、中、高剂量组。CCK-8法分别检测不同浓度ox-LDL和黄芪赤风汤对bEnd.3细胞活力的影响；油红O染色评估bEnd.3细胞内脂质聚积情况；酶联免疫吸附试验（ELISA）法检测bEnd.3细胞总胆固醇（TC）、游离胆固醇（FC）、胆固醇酯（CE）、白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)、细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)水平和CE/TC；非靶向代谢组学分析bEnd.3细胞的代谢物谱，以鉴定筛选出与内皮损伤相关的差异代谢物，从而探究黄芪赤风汤保护内皮损伤的代谢通路及潜在作用机制。结果 CCK-8结果确定50μg/mL为ox-LDL的造模浓度，50、100和200μg/mL为黄芪赤风汤干预bEnd.3细胞的低、中、高剂量。与模型组比较，黄芪赤风汤能有效改善bEnd.3细胞的泡沫化程度，脂滴蓄积明显减少，黄芪赤风汤各剂量组TC、CE和TNF-α水平显著下调（P<0.05、0.01），黄芪赤风汤中、高剂量组FC、IL-6、VCAM-1水平和CE/TC显著下调（P<0.05、0.01），黄芪赤风汤高剂量组IL-1β和ICAM-1水平显著下调（P<0.05、0.01）。代谢组学鉴定出15种差异代谢产物，包括5-羟基吲哚乙醛、同型半胱氨酸、花生四烯酸、鞘氨醇-1-磷酸、苯丙酮酸、谷胱甘肽、氧化谷胱甘肽和L-磷酸精氨酸等，主要富集于谷胱甘肽代谢、烟酸和烟酰胺代谢、苯丙氨酸代谢、半胱氨酸和蛋氨酸代谢等代谢途径。结论 黄芪赤风汤可有效改善内皮细胞损伤，抑制ox-LDL诱导的bEnd.3细胞内脂质蓄积、炎性因子和黏附分子表达，其机制可能与调节受损内皮细胞内的代谢功能，调节细胞内氨基酸等物质代谢相关。

Objective To investigate the protective effect of Huangqi Chifeng Tang on endothelial cell injury and its mechanism based on metabolomics. Methods An in vitro endothelial cell injury model was established by ox-LDL induced mouse brain endothelial bEnd.3 cells. bEnd.3 cells were randomly divided into control group, model group and Huangqi Chifeng Tang low, medium, and high dose groups. Cell counting Kit-8（CCK-8） was used to detect the effects of different concentrations of ox LDL and Huangqi Chifeng Tang on bEnd.3 cell viability. Oil red O staining was used to evaluate the intracellular lipid accumulation in bEnd.3 cells. Enzymelinked immunosorbent assay（ELISA） was used to detect the levels of total cholesterol（TC）, free cholesterol（FC）, cholesterol esters（CE）, interleukin-1β（IL-1β）, IL-6, tumor necrosis factor-α（TNF-α）, intercellular adhesion molecule-1（ICAM-1）, and vascular cell adhesion molecule-1（VCAM-1） in bEnd.3 cells, as well as CE/TC. Non-targeted metabolomics analysis of the metabolite profile of bEnd.3 cells to identify and screen differential metabolites related to endothelial injury, in order to explore the metabolic pathways and potential mechanisms of Huangqi Chifeng Tang in protecting endothelial injury. Results CCK-8 determined 50 μg/mL as the modeling concentration for ox-LDL, and 50, 100, and 200 μg/mL were the low, medium, and high dose of Huangqi Chifeng Tang intervention in bEnd.3 cells. Compared with the modal group, Huangqi Chifeng Tang can effectively improve the foam degree of bEnd.3 cells, and significantly reduced the accumulation of lipid droplets. Compared with the modal group, the levels of TC, CE, and TNF-α were significantly decreased in each dose group of Huangqi Chifeng Tang（P＜0.05, 0.01）, the levels of FC, IL-6, VCAM-1, and CE/TC were significantly decreased in the middle and high dose groups of Huangqi Chifeng Tang（P＜0.05, 0.01）, and the levels of IL-1β and ICAM-1 were significantly decreased in the high dose group of Huangqi Chifeng Tang（P＜0.05, 0.01）. Metabolomics results showed that a total of 15 differential metabolites were identified, including 5-hydroxyindolealdehyde, homocysteine, arachidonic acid, sphingosine-1-phosphate, phenylpyruvic acid, glutathione, oxidized glutathione and L-Phosphoarginine, mainly enriched in metabolic pathways such as glutathione metabolism, niacin and nicotinamide metabolism, phenylalanine metabolism, cysteine and methionine metabolism. Conclusion Huangqi Chifeng Tang can effectively improve endothelial cell damage, inhibit ox-LDL induced lipid accumulation, inflammatory factors, and adhesion molecule expression in bEnd. 3 cells. Its mechanism may be related to regulating the metabolic function of damaged endothelial cells and regulating the metabolism of intracellular amino acids and other substances.

R285.5

黑龙江省自然科学基金资助项目(PL2024H239)