糖尿病（DM）及其并发症严重威胁人类的生命和健康。 Rhaponticum cartharoides (Wild.) Iljin (RC) 广泛用于治疗心血管疾病。先前的研究报告称，RC 可以降低 1 型糖尿病大鼠的血糖水平。然而，RC对2型糖尿病和血管并发症的影响及其相关活性成分和潜在机制仍不清楚。本研究旨在探讨RC对2型糖尿病小鼠和糖尿病患者内皮功能障碍和炎症反应的影响。以雄性C57BL/6J小鼠建立2型糖尿病小鼠模型。给小鼠口服 RC 提取物（60、120 和 240 毫克/千克）12 周后，评估血糖和血脂水平。采用苏木精和伊红（HE）染色观察肝脏和肾脏组织的形态结构，并通过检测血清中的相关生化指标评价其功能。然后通过HE染色观察主动脉形态。此外，检测血清内皮功能标志物、炎症因子的含量，并检测主动脉炎症因子的表达量及关键蛋白的磷酸化水平。此外，利用各种数据库进行药效团匹配的基础上，对RC作用于糖尿病血管病变的潜在靶点进行预测和富集分析。然后，使用蛋白质印迹、免疫荧光和 RT-PCR 评估接受 RC 治疗的 DM 小鼠主动脉中潜在靶标的表达、定位和磷酸化水平。最后，通过虚拟筛选确定了RC的活性成分，并验证了其改善内皮细胞功能障碍的能力。RC降低了血糖水平以及总甘油三酯（TG）、总胆固醇（TC）和低密度脂蛋白的血脂水平胆固醇（LDL-c），增加高密度脂蛋白胆固醇（HDL-c）水平，并改善 2 型糖尿病小鼠的肝肾功能。 RC 减少 2 型 DM 小鼠主动脉内皮细胞脱落，增加血清一氧化氮 (NO) 和一氧化氮合酶 (NOS) 水平，并减少可溶性分化簇 40 配体 (sCD40L)、肿瘤坏死因子 α (TNF-α) ) 和白介素-1β (IL-1β) 水平。进一步的研究结果表明，RC 降低了主动脉炎症因子，即 CD40、CD40L、IL-1β 和白细胞介素 6 (IL-6) 的表达，并增加了内皮型一氧化氮合酶 (eNOS) 磷酸化水平。 Sirtuin 6（SIRT6）、蛋白激酶B（AKT）和eNOS被预测是RC作用于DM血管病变的关键节点靶标，并且证实RC增加了主动脉内皮细胞中SIRT6的表达和AKT磷酸化水平。 20-羟基蜕皮酮 (20E)、胡萝卜素 (Dau)、桉树酸 (Eus) 和丁香苷 (Syr) 被确定为 RC 的活性成分。这些成分可防止 TNF-α 诱导的人脐静脉内皮细胞 (HUVEC) 损伤，并减少乳酸脱氢酶 (LDH) 和 IL-1β 的释放，并以剂量​​依赖性方式增加 TNF-α 诱导的 HUVEC 中 NO 的释放。 RC降低2型糖尿病小鼠的血糖和血脂水平，并保护肝肾功能。 RC促进内皮细胞SIRT6表达；通过增加 AKT/eNOS 磷酸化水平来上调 NO/NOS 系统，从而调节血管张力因子；降低CD40、TNF-α、IL-1β等炎症因子水平，抑制内皮炎症反应。基于这些机制，RC 可以改善内皮功能障碍。版权所有 © 2024 Elsevier GmbH。版权所有。

Diabetes mellitus (DM) and its complications seriously threaten human life and health. Rhaponticum carthamoides (Willd.) Iljin (RC) is widely used to treat cardiovascular diseases. Previous studies reported that RC reduces blood glucose levels in rats with type 1 DM. However, the effects of RC on type 2 diabetes and vascular complications, as well as its related active components and underlying mechanisms, remain unclear.This study aimed to investigate the effects of RC on endothelial dysfunction and the inflammatory response in type 2 DM mice and to explore its underlying mechanism and active ingredients.Male C57BL/6J mice were used to establish a type 2 DM mouse model. After 12 weeks of oral administration of RC extract (60, 120, and 240 mg/kg) to mice, blood glucose and lipid levels were assessed. The morphological structures of the liver and kidney tissues were observed using hematoxylin and eosin (HE) staining, and their functions were evaluated by detecting relevant biochemical indicators in the serum. Then, aorta morphology was observed via HE staining. In addition, serum levels of markers of endothelial function and inflammatory factors were detected, and the expression of inflammatory factors and the phosphorylation levels of key proteins in the aorta were examined. Furthermore, prediction and enrichment analyses of potential targets of RC acting on diabetic vascular lesions were performed on the basis of pharmacophore matching using various databases. Then, the expression, localization and phosphorylation levels of potential targets in the aortas of DM mice treated with RC were assessed using Western blotting, immunofluorescence, and RT‒PCR. Finally, the active components of RC were identified through virtual screening, and their ability to improve endothelial cell dysfunction was verified.RC reduced blood glucose levels and serum lipid levels of total triglyceride (TG), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-c), increased high density lipoprotein cholesterol (HDL-c) levels, and improved liver and kidney function in type 2 DM mice. RC decreased endothelial cell shedding in the aortas of type 2 DM mice, increased serum nitric oxide (NO) and nitric oxide synthase (NOS) levels, and reduced soluble cluster of differentiation 40 ligand (sCD40L), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) levels. Further findings indicated that RC reduced the expression of aortic inflammatory factors, namely, CD40, CD40L, IL-1β, and interleukin-6 (IL-6), and increased endothelial NOS (eNOS) phosphorylation levels. Sirtuin 6 (SIRT6), protein kinase B (AKT), and eNOS were predicted to be key node targets of RC acting on DM vascular lesions, and it was confirmed that RC increased SIRT6 expression and AKT phosphorylation levels in aortic endothelial cells. 20-Hydroxyecdysone (20E), daucosterol (Dau), euscaphic acid (Eus), and syringin (Syr) were identified as active components of RC. These components protect against TNF-α-induced human umbilical vein endothelial cell (HUVEC) damage and decrease the release of lactate dehydrogenase (LDH) and IL-1β and increased the release of NO in TNF-α-induced HUVECs in a dose-dependent manner.RC reduced blood glucose and lipid levels in mice with type 2 DM and protected liver and kidney function. RC promotes SIRT6 expression in endothelial cells; upregulates the NO/NOS system by increasing AKT/eNOS phosphorylation levels to regulate vascular tone factors; and reduces the levels of inflammatory factors such as CD40, TNF-α, and IL-1β to inhibit endothelial inflammatory responses. Based on these mechanisms, RC improves endothelial dysfunction.Copyright © 2024 Elsevier GmbH. All rights reserved.