南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (8): 1425-1431.doi: 10.12122/j.issn.1673-4254.2023.08.21

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丹参新醌乙减轻ox-LDL诱导的内皮细胞损伤：基于抑制NF-κB/NLRP3信号通路介导的细胞焦亡

李洪涛，邓 宇，王添乐，黄克勇，于传沛，陈朝俊   

  1. 广州中医药大学，广东 广州 510006；连山县小三江镇中心卫生院，广东 清远 513224；广州中医药大学附属广州中西医结合医院脑病科，广东 广州 510800
  • 出版日期:2023-08-20 发布日期:2023-09-13

Danshenxinkun B protects human umbilical vein endothelial cells against ox-LDL-induced injury by inhibiting pyroptosis and the NF-κB/NLRP3 pathway

LI Hongtao, DENG Yu, WANG Tianle, HUANG Keyong, YU Chuanpei, CHEN Chaojun   

  1. Guangzhou University of Chinese Medicine, Guangzhou 510006, China; Township Center Hospital of Xiaosanjiang Town, Qingyuan 513224, China; Department of Neurology, Guangzhou Hospital of Integrated Traditional and West Medicine Affiliated to Guangzhou University of Chinese Medicine, Guangzhou 510800, China
  • Online:2023-08-20 Published:2023-09-13

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摘要： 目的 探讨丹参新醌乙对氧化低密度脂蛋白（ox-LDL）诱导的人脐静脉内皮细胞损伤的影响及机制。方法 取对数生长期的人脐静脉内皮细胞，分为正常组（加入10%胎牛血清）、ox-LDL组（在正常组基础上加入100ug/mL的ox-LDL）、二甲基亚砜（DMSO）组（在ox-LDL组基础上加入0.1%浓度DMSO）、丹参新醌乙组（在ox-LDL组基础上加入由DMSO溶解的100ng/ml的丹参新醌乙），培养24 h后进行实验。微板法检测乳酸脱氢酶（LDH）表达；qRT-PCR法检测NF-κB1、核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）、消皮素D（GSDMD）、白介素（IL）-1β的mRNA表达；Western Blot法检测NF-κB1、NLRP3、caspase-1、GSDMD-N的蛋白表达。免疫荧光法检测GSDMD表达情况。结果 与正常组比较，ox-LDL组、DMSO组LDH表达明显升高（P<0.01），NF-κB1、NLRP3、GSDMD、IL-1β的mRNA水平均显著上调（P<0.01），NF-κB1、NLRP3、caspase-1、GSDMD-N、IL-1β的蛋白表达水平明显升高（P<0.01）；与 ox-LDL 组、DMSO 组比较，丹参新醌乙组 LDH 表达有所降低（P<0.05），NF-κB1、NLRP3、GSDMD、IL-1β的mRNA水平显著下调（P<0.01），NF-κB1、NLRP3、caspase-1、GSDMD-N、IL-1β的蛋白水平显著降低（P<0.01），GSDMD在质膜表达抑制，入核受限。结论 丹参新醌乙可能通过NF-κB/NLRP3信号通路，抑制NLRP3炎症小体介导的细胞焦亡，改善ox-LDL诱导的内皮细胞损伤。

关键词: NF-κB/NLRP3信号通路；细胞焦亡；丹参新醌乙；氧化低密度脂蛋白；内皮细胞损伤；炎症；动脉粥样硬化

Abstract: Objective To investigate the protective effect of Danshenxinkun B against oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cell (HUVEC) injury and explore the underlying mechanism. Methods HUVECs cultured in the presence of 10% fetal bovine serum were treated with ox-LDL (100 μg/mL), ox-LDL+0.1% dimethyl sulfoxide (DMSO), or ox-LDL+Danshenxinkun B (100 ng/mL, dissolved in DMSO) for 24 h. The changes in lactate dehydrogenase (LDH) release was detected, and qRT-PCR was used to detect the mRNA expressions of nuclear factor-κB1 (NF-κB1), nucleotide binding oligomerization domain-like receptor family pyrin domain protein 3 (NLRP3), gasdermin D (GSDMD) and interleukin-1β (IL-1β). The protein expressions of NF-κB1, NLRP3, caspase-1, IL-1β and GSDMD-N were detected with Western blotting. Immunofluorescence assay was performed to examine the changes in GSDMD expression in the cells. Results Compared with the normal control cells, the cells treated with ox-LDL alone or in combination with DMSO exhibited significantly increased LDH release, mRNA expressions of NF-κB1, NLRP3, GSDMD, and IL- 1β and the protein levels of NF- κB1, NLRP3, IL-1β, GSDMD-N and caspase-1 (P<0.01), which were all significantly lowered by treatment with Danshenxinkun B (P<0.05 or 0.01). Danshenxinkun B treatment significantly inhibited GSDMD expression on the cell membrane and restricted its entry into the cell nucleus. Conclusion Danshenxinkun B alleviates ox-LDL-induced HUVEC injury possibly by suppressing pyroptosis mediated by NLRP3 inflammatory bodies and inhibiting the NF-κB/NLRP3 signaling pathway

Key words: NF-κB/NLRP3 signaling pathway; pyroptosis; Danshenxinkun B; oxidized low-density lipoprotein; endothelial cell injury; inflammation; therosclerosis