苦杏仁苷减少冠状动脉内皮细胞焦亡并改善载脂蛋白E缺陷小鼠动脉粥样硬化斑块形成  

Amygdalin reduces coronary endothelial pyroptosis and ameliorates atherosclerotic plaque formation in ApoE^(-/-) mice

作　　者：刘瑾[1] 张洁[1] 冯莹[1] LIU Jin;ZHANG Jie;FENG Ying(Department of Cardiovascular Medicine,Wuhan First Hospital,Wuhan 430000,China)

机构地区：[1]武汉市第一医院心血管内科,武汉430000

出　　处：《实用医学杂志》2023年第14期1746-1755,共10页The Journal of Practical Medicine

基　　金：武汉卫健委青年科学基金(编号:WX21Q11)。

摘　　要：目的研究苦杏仁苷(amygdalin,AMY)对动脉粥样硬化的治疗作用及其可能的作用机制。方法利用不同剂量AMY干预人冠状动脉内皮细胞(HCAECs)。将HCAECs分为4组:对照组、ox-LDL组、AMY低剂量组(50μg/mL)和AMY高剂量组(100μg/mL)。Western blot检测焦亡标志蛋白(Caspase-1和GSDMD)以及组蛋白去甲基化酶(JMJD3)和半乳糖凝集素-3(Gal-3)表达水平;RT-qPCR检测IL-1β和IL-18 mRNA表达;TUNEL测试检测HCAECs死亡率。ChIP试验用于检测JMJD3和H3K27me3在Gal-3启动子上的富集。过表达Gal-3后分析Gal-3对AMY抑制ox-LDL诱导的HCAECs焦亡的影响。将40只载脂蛋白E缺陷(ApoE-/-)雄性小鼠分为对照组、模型组、2.5 mg/kg和5 mg/kg AMY组。油红O染色用于观察小鼠主动脉脂质沉积和斑块形成;酶比色法测定血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)含量。结果与对照组比较,AMY组对细胞存活率无显著影响,50、100μg/mL AMY干预细胞24 h内细胞存活率与对照相比差异无统计学意义,36、48 h时细胞存活率相较对照组降低;与ox-LDL比较,50和100μg/mL AMY组Caspase-1、GSDMD、Gal-3和JMJD3蛋白水平,IL-1β和IL-18 mRNA表达以及TUNEL阳性细胞率呈剂量依赖性降低;AMY干预后HCAECs内Gal-3启动子上JMJD3富集降低,H3K27me3富集升高;与ox-LDL+AMY+pcDNA-3.1组比较,ox-LDL+AMY+pcDNA-Gal-3组Gal-3、Caspase-1、GSDMD、IL-1β、IL-18表达水平和TUNEL阳性细胞率明显增加;与模型组相比,2.5 mg/kg AMY组、5 mg/kg AMY组小鼠中主动脉中脂质沉积显著减少,动脉粥样硬化病变面积百分比降低;TG、TC、LDL浓度和Gal-3、Caspase-1、GSDMD、IL-1β、IL-18表达水平呈剂量依赖性降低,HDL含量升高。结论AMY可能通过参与调控JMJD3介导的Gal-3去甲基化对动脉粥样硬化导致的细胞焦亡有一定的改善作用。Objective To investigate the therapeutic effects of amygdalin(AMY)on atherosclerosis and to elucidate its possible mechanisms.Methods Treatment with different doses of AMY intervention on human coronary artery endothelial cells(HCAECs),HCAECs were divided into four groups:control,ox-LDL,AMY low dose group(50μg/mL),and AMY high dose group(100μg/mL).Pyroptosis proteins(caspase-1 and GSDMD)as well as histone demethylase(JMJD3)and galectin-3(Gal-3)expression levels were determined by Western blotting.IL-1βand IL-18 mRNA expression by RT-qPCR.TUNEL assay was used to detect the death rate of HCAECs.ChIP assay was used to detect the enrichment of JMJD3 and H3K27me3 on Gal-3 promoter region.The effect of Gal-3 on Amy inhibition of ox LDL induced pyroptosis in HCAECs was further analyzed after overexpression of Gal-3.40 apolipo protein E-deficient(ApoE)-/-male mice were divided into control group,model group,2.5 and 5 mg/kg AMY group.Oil red O staining was used to observe lipid deposition and plaque formation in mice aortas.Serum levels of total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)were measured by enzymatic colorimetric methods.Results Versus the control group,AMY had no significant effect on cell viability,50 and 100μg/mL AMY intervened cells showed no significant difference in cell viability compared to the control at 24 h,and a significant decrease in cell viability compared to the control at 36 h and 48 h.Compared to ox-LDL,the protein levels of Caspase-1,GSDMD,Gal-3 and JMJD3,the mRNA expression of IL-1βand IL-18,and the rate of TUNEL positive cells decreased in a dose-dependent manner 50 and 100μg/mL AMY.The enrichment of JMJD3 on Gal-3 promoter region was decreased and H3K27me3 was increased in HCAECs after AMY intervention.The expression levels of Gal-3,caspase-1,GSDMD,IL-1β,and IL-18 and the percentage of TUNEL positive cells within HCAECs were increased in the ox-LDL+AMY+pcDNA-Gal-3 group compared with the ox-LDL+AMY+p

关 键 词：动脉粥样硬化 细胞焦亡 苦杏仁苷 JMJD3 GAL-3 

分 类 号：R96[医药卫生—药理学] R541.4[医药卫生—药学]