炎症在动脉粥样硬化（AS）的病理生理中起着重要作用。本研究的目的是调查丹酚酸B（Sal B）对AS的保护作用，以及肿瘤坏死因子-α（TNF-α）诱导人脐静脉内皮细胞（HUVECs）损伤的分子机制。在体研究中，采用高脂饮食（HFD）喂养LDLR-/-小鼠14周，建立AS模型，以评估Sal B对AS发展的保护作用。测定血清中的总胆固醇（TC）、甘油三酯（TG）和低密度脂蛋白胆固醇（LDL-C）水平。使用油红O染色测量并定量表面和横截面脂质沉积物。使用血红染和伍氏染色测量主动脉根部切片中的动脉粥样硬化斑块大小和胶原纤维含量。使用二氢乙二胺（DHE）染色检测主动脉根部的活性氧（ROS）释放。通过TdT介导的dUTP末端标记（TUNEL）染色确定细胞凋亡率。使用免疫荧光（IF）染色检测核因子κ-B（NF-κB）p65和NOD样受体家族簇鸟神经领域含有3（NLRP3）的表达。为进一步研究Sal B的保护作用，我们使用TNF-α诱导的HUVECs炎症模型。我们检测了细胞存活率、乳酸脱氢酶（LDH）含量和ROS产生。通过免疫荧光评估了NF-κB的转录。通过RT-PCR检测了NLRP3、半胱氨酸蛋白酶-1（caspase-1）和IL-1β的mRNA水平。通过Western blot检测了金属蛋白酶ROS后的相关蛋白。小鼠体重随时间的变化表明，Sal B对体重增加有影响。我们能够显示出模型组血清中的TC、TG和LDL-C增加，而Sal B治疗降低了血清脂质水平。组织学染色结果显示，LDLR-/-小鼠在MOD组中有大量泡沫细胞沉积物，伴有炎症细胞浸润和动脉粥样硬化斑块形成。Sal B治疗显著改善了这些病理异常。HFD后主动脉根部的ROS释放和细胞凋亡显著增加，在Sal B治疗下得到了缓解。IF结果显示，NF-κB p65和NLRP3的表达在MOD组显著增加，在Sal B组显著减少，表明Sal B可能通过NF-κB/NLRP3途径发挥作用。并且在体外研究中，通过TNF-α诱导HUEVCs的炎症损伤，在Sal B处理后，细胞存活率显著增加，并且LDH释放减少。还发现Sal B抑制了TNF-α诱导的HUEVCs后ROS水平的增加，通过NF-κB p65的激活，NF-κB p65转移到细胞核。Sal B处理可以逆转这种效应。RT-PCR和Western blot显示，Sal B影响NF-κB的转录和NLRP3炎性小体的激活，并能明显抑制TNF-α诱导的NLRP3炎性小体的激活。这些结果表明，Sal B可能通过NF-κB/NLRP3途径参与抗动脉粥样硬化和炎症反应。本研究显示，Sal B改善了HFD诱导的LDLR-/-小鼠AS病变的发展。此外，在TNF-α条件下，Sal B减少ROS的释放，逆转了NF-κB的核内转运，并通过调控NF-κB/NLRP3途径抑制动脉粥样硬化和炎症。版权所有 © 2023 Elsevier GmbH。保留所有权利。

Inflammation is critical in the pathophysiology of atherosclerosis (AS). The aim of this study was to investigate the protective effect of salvianolic acid B (Sal B) on AS and to explore the molecular mechanism of tumor necrosis factor-α (TNF-α)-induced damage in human umbilical vein endothelial cells (HUVECs).In vivo studies, LDLR-/- mice were fed a high-fat diet (HFD) for 14 weeks to establish an AS model to evaluate the protective effect of Sal B on the development of AS. Total cholesterol (TC), triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) levels were determined in the blood serum. En face and cross section lipid deposits were measured and quantified with Oil Red O staining. Hematoxylin and eosin (H&E) and Masson's trichrome staining were used to quantify atherosclerotic plaque size and collagen fiber content in aortic root sections. Reactive oxygen species (ROS) were detected in aortic root using dihydroethylenediamine (DHE) staining. Apoptosis rate was determined by TdT-mediated dUTP nick end labeling (TUNEL) staining. Immunofluorescence (IF) staining was used to detect the expression of the nuclear factor kappa-B (NF-κB) p65 and NOD-like receptor family pyrin domain containing 3 (NLRP3). To further investigate the protective effect of Sal B, we used TNF-α induced HUVECs inflammation model. We examined cell viability, lactate dehydrogenase (LDH) content, and ROS production. The transcription of NF-κB was evaluated by immunofluorescence. The mRNA levels of NLRP3, caspase-1, and IL-1β were detected by RT-PCR. Pyroptosis related proteins were detected by Western blot.The change in the weight of the mice over time was an indication that Sal B had an effect on weight gain.we were able to show that the serum lipids TC, TG and LDL-C were increased in the model group and that the treatment with Sal B reduced the levels of serum lipids. Histological staining showed that the LDLR-/- mice had a large amount of foam cell deposition accompanied by inflammatory cell infiltration and the formation of atherosclerotic plaques in theMOD group. The pathological abnormalities were significantly improved by Sal B treatment. ROS release and apoptosis were significantly increased after HFD in aortic root, which was attenuated by Sal B. IF results showed that the expression of NF-κB p65 and NLRP3 was significantly increased in the MOD group and significantly decreased in the Sal B group, suggesting that Sal B may act through the NF-κB/NLRP3 pathway. And in vitro studies: inflammatory damage of HUEVCs was induced by TNF-α, and Sal B treatmented significantly increased cell viability and reduced LDH release. It was also found that Sal B inhibited ROS level increase after TNF-α-induced HUEVCs. Activation of NF-κB p65 by TNF-α stimulation, NF-κB p65 is transferred to the nucleus. Sal B treatment could reverse this effect. RT-PCR and Western blot showed that Sal B affected NF-κB transcription and NLRP3 inflammasome activation and could significantly inhibit TNF-α-induced NLRP3 inflammasome activation. These results suggest that Sal B may participate in antiatherosclerotic and inflammatory responses through the NF-κB/NLRP3 pathway.This study shows that Sal B ameliorates the development of AS lesions in HFD-induced LDLR-/- mice. Furthermore, under TNF-α conditions, Sal B reduced ROS release and reversed nuclear translocation of NF-κB, and inhibited atherosclerosis and inflammation by modulating the NF-κB/NLRP3 pathway.Copyright © 2023 Elsevier GmbH. All rights reserved.