槲皮素通过抑制小鼠骨髓来源泡沫细胞焦亡抗动脉粥样硬化的研究

Quercetin against atherosclerosis by inhibiting pyroptosis of bone marrow-derived foam cells in mice

目的本研究探讨了槲皮素对骨髓来源泡沫细胞焦亡的拮抗作用,从抗焦亡角度研究槲皮素抗动脉粥样硬化的作用和机制。方法(1)动物实验:槲皮素灌胃动脉粥样硬化模型小鼠进行分组,油红O染色检测槲皮素对小鼠动脉粥样硬化病变的作用;蛋白质印迹法检测动脉粥样硬化组织中焦亡相关蛋白的表达。(2)细胞实验:培养原代小鼠骨髓来源巨噬细胞(BMDM),应用ox-LDL诱导泡沫细胞形成,应用细胞活力实验检测槲皮素作用于小鼠骨髓来源泡沫细胞的最佳浓度;酶联免疫吸附实验(ELISA)检测培养基中乳酸脱氢酶(LDH)量;流式细胞术检测细胞中活性氧(ROS)量;蛋白质印迹法检小鼠骨髓来源泡沫细胞中焦亡相关蛋白的表达。结果高浓度槲皮素干预后动脉粥样硬化模型小鼠动脉窦区油红O染色动脉粥样硬化减轻,动脉粥样硬化病变组织中焦亡相关蛋白下调,一氧化氮(NO)量下调。槲皮素作用于小鼠骨髓来源泡沫细胞后培养基中LDH量下降,蛋白质印迹显示焦亡相关蛋白下调。结论槲皮素可能通过抑制ROS产生下调NLRP3的表达,进而抑制泡沫细胞的焦亡,改善动脉粥样硬化。

Objective To investigate the antagonistic effect of quercetin on bone marrow-derived foam cell pyroptosis,and to study the anti-atherosclerosis effect and mechanism of quercetin from the perspective of anti-pyroptosis.Methods In animal experiments,quercetin gavage atherosclerosis model mice were grouped,and the effect of quercetin on atherosclerotic lesions in mice was detected by oil red O staining,and the expression of pyroptosis-related proteins in atherosclerotic tissues was also detected by Western blotting.In cell experiments,primary mouse bone marrow-derived macrophages(BMDMs)were cultured,ox-LDL was used to induce the formation of foam cells,cell viability assay to detect the optimal concentration of quercetin on mouse bone marrow-derived foam cells,enzyme-linked immunosorbent assay(ELISA)to detect the amount of lactate dehydrogenase(LDH)in the medium,flow cytometry to detect the amount of reactive oxygen species(ROS)in cells,and the expression of pyroptosis-related protein in mouse bone marrow-derived foam cells was detected by Western blotting.Results After high-concentration quercetin intervention,the atherosclerosis of the arterial sinus stained with oil red O was reduced,the pyroptosis-related protein and the amount of nitric oxide(NO)in atherosclerotic lesions were down-regulated.The amount of LDH in the medium decreased after quercetin was applied to mouse bone marrow-derived foam cells,and Western blotting showed that pyroptosis-related proteins were down-regulated.Conclusion Quercetin may down-regulate the expression of NLRP3 by inhibiting ROS production,thereby inhibiting pyroptosis of foam cells and improving atherosclerosis.