目的 探讨白藜芦醇对动脉粥样硬化（AS）中血管钙化的影响及机制。方法 将雄性健康SD大鼠随机分为对照组、模型组、白藜芦醇（5 mg·kg^-1）组，每组10只。造模前ip预给药21 d，每天1次。模型组、白藜芦醇组均制备AS模型：sc 5 mg·kg^-1维生素D₃5 d后，分离并结扎左颈动脉，0.12 mmol·L^-1的CaCl₂溶液湿敷30 min，随即缝合，对照组湿敷生理盐水。造模后继续给药，1周后所有大鼠同时禁食24 h后处死，取大鼠左颈动脉进行苏木精-伊红（HE）染色和Von Kossa染色。体外钙化培养基诱导CRL-1999细胞钙化，给予白藜芦醇（10μmol·L^-1）干预，茜素红S染色检测钙盐沉积；实时荧光定量PCR （qRT-PCR）和Western blotting法检测钙化指标骨形态发生蛋白-2（BMP2）、侏儒相关转录因子2（Runx2），炎性小体NOD样受体热蛋白结构域相关蛋白3（NLRP3），细胞焦亡相关指标天冬氨酸蛋白水解酶-1（Caspase-1）、Gasdermin-D （GSDMD）、白细胞介素（IL）-1β、IL-18蛋白和mRNA表达变化；免疫荧光检测Runx2、NLRP3、Caspase-1、GSDMD、IL-1β蛋白表达；分子对接验证白藜芦醇与靶蛋白NLRP3、Caspase-1、GSDMD、BMP2、Runx2结合活性。结果 HE与VonKossa染色显示模型组血管壁结构紊乱、钙盐沉积，白藜芦醇缓解钙盐沉积。体外实验表明钙化结节可被白藜芦醇缓解；与对照组比较，模型组BMP2、Runx2、NLRP3、Caspase-1、GSDMD、IL-1β、IL-18 mRNA和蛋白表达均显著上升（P<0.05、0.01、0.001）；经白藜芦醇处理后，上述指标的mRNA和蛋白表达均显著下降（P<0.05、0.01、0.001）。白藜芦醇与Runx2、NLRP3分别形成2、3个氢键，对接活性较好。结论 白藜芦醇抑制AS中血管钙化，机制可能与抑制NLRP3/Caspase-1信号通路，进而抑制细胞焦亡有关。

Objective To investigate the effect of resveratrol on vascular calcification in atherosclerosis(AS) and its mechanism.Methods Male healthy SD rats were randomly divided into control group, model group and resveratrol(5 mg·kg^-1) group, with 10 rats in each group.Resveratrol was ip preadministered for 21 days, once a day, before modeling.AS model was prepared in the model group and the resveratrol group:sc 5 mg·kg^-1 vitamin D3 for five days, the left carotid artery was isolated and ligated, and0.12 mmol·L^-1 CaCl₂ solution was wet applied for 30 min, and then sutured.The control group was wet applied normal saline.One week later, all rats were sacrificed after fasting for 24 h at the same time.The left carotid arteries of the rats were taken for HE staining and Von Kossa staining.The calcification of CRL-1999 cells was induced by calcification medium in vitro, and resveratrol(10 μmol·L^-1) was administered.Alizarin red S staining was used to detect calcium salt deposition.The expression of calcium markers BMP2, Runx2, inflammasome NLRP3, pyroptosis related indicators aspartic acid proteolytic enzyme 1(Caspase-1), Gasdermin-D(GSDMD), interleukin(IL)-1β and IL-18 were detected by Quantitative real-time PCR(qRT-PCR) and Western blotting.The expressions of Runx2, NLRP3, Caspase-1, GSDMD and IL-1β were detected by immunofluorescence.The binding activity of resveratrol to target protein was verified by molecular docking.Results HE and Von Kossa staining showed vascular wall structure disorder and calcium salt deposition in the model group, which was alleviated by resveratrol pretreatment.VSMCs in vitro showed that calcified nodules could be alleviated by resveratrol.The expressions of BMP2, Runx2, inflammasome NLRP3, Caspase-1, GSDMD, IL-1β and IL-18 in model group were increased(P<0.05, 0.01 and 0.001).After resveratrol treatment, the mRNA and protein expressions of the above indexes were decreased(P<0.05, 0.01 and 0.001).Resveratrol formed two and three hydrogen bonds with Runx2 and NLRP3, respectively, showing good docking activity.Conclusion Resveratrol inhibits vascular calcification in AS, and the mechanism may be related to the inhibition of NLRP3/Caspase-1 signaling pathway and the inhibition of pyroptosis.

R285.5

国家自然科学基金面上项目（8207131747）；湖北省卫生健康委中医药科研立项项目（ZY2021F007）