葛根素通过抑制ox-LDL诱导巨噬细胞焦亡通路活化稳定AS易损斑块  

Puerarin inhibits ox-LDL induces macrophage pyrolytic pathway activation to stabilize AS vulnerable plaque

作　　者：章平衡 李春燕 刘永源[1] 王明[1] 刘锦鸿 梁东辉[1] ZHANG Ping-Heng;LI Chun-Yan;LIU Yong-Yuan;WANG Ming;LIU Jin-Hong;LIANG Dong-Hui(Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China)

机构地区：[1]南方医科大学珠江医院,广州510282

出　　处：《中国免疫学杂志》2021年第18期2212-2216,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(81774036)。

摘　　要：目的:研究葛根素对ox-LDL诱导巨噬细胞NLRP3/Caspase-1焦亡信号通路的影响,探讨葛根素稳定动脉粥样硬化(AS)易损斑块的潜在调节机制。方法:采用佛波酯(PMA)刺激THP-1细胞48 h,诱导其分化为巨噬细胞,分别给予ox-LDL(100μg/ml)、低(100μg/ml)、中(250μg/ml)和高剂量葛根素(500μg/ml)。CCK-8法检测不同浓度葛根素对巨噬细胞存活率的影响。油红O染色检测巨噬细胞内脂质聚集及其泡沫化情况;RT-qPCR检测炎症细胞因子IL-6、TNF-α、IL-1β及IL-2 mRNA水平;Western blot检测细胞焦亡蛋白NLRP3、Caspase-1、cleaved-Caspase-1水平,细胞免疫荧光法测定NLRP3表达和定位。结果:高、中、低剂量葛根素对巨噬细胞存活率无明显影响(P>0.05),均可抑制脂质在巨噬细胞内沉积及泡沫化(P<0.01),下调炎症细胞因子IL-6、TNF-α、IL-1β、IL-2 mRNA水平(P<0.01),抑制细胞焦亡蛋白NLRP3、cleaved-Caspase-1表达(P<0.01),并呈剂量依赖性。结论:葛根素可能通过抑制ox-LDL诱导的巨噬细胞NLRP3/Caspase-1焦亡信号通路活化,下调炎症细胞因子表达,稳定AS易损斑块。Objective:To investigate effect of puerarin on ox-LDL induced focal death signaling pathway NLRP3/Caspase-1 of macrophages,and to explore potential regulatory mechanism of puerarin stabilizing atherosclerotic(AS)vulnerable plaque.Methods:THP-1 cells were stimulated by phorbol ester(PMA)for 48 h to induce them differentiation into macrophages,and intervened by ox-LDL(100μg/ml),low dose(100μg/ml),medium dose(250μg/ml)and high dose puerarin(500μg/ml),respectively.Effects of different concentrations of puerarin on survival rate of macrophages were detected by CCK-8.Lipid aggregation and foaming in macrophages were detected by oil red O staining.Inflammatory cytokine IL-6,TNF-α,IL-1βand IL-2 mRNA levels were detected by RTqPCR.Protein expressions of NLRP3,Caspase-1,cleaved-Caspase-1 were detected by Western blot,protein expression and localization of NLRP3 were detected by cell immunofluorescence.Results:High,medium,low doses of puerarin had no significant effect on macrophage survival(P>0.05),and could inhibit lipid deposition and foamation in macrophages(P<0.01),decrease levels of inflammatory cytokine IL-6,TNF-α,IL-1β,IL-2 mRNA(P<0.01),and inhibit protein expressions of NLRP3,cleaved-Caspase-1(P<0.01),which in a dose-dependent manner.Conclusion:Puerarin may stabilize AS vulnerable plaque by inhibiting ox-LDL-induced activation of NLRP3/Caspase-1 focal death signaling pathway forehead and down-regulating inflammatory cytokines expressions.

关 键 词：动脉粥样硬化 葛根素 巨噬细胞 焦亡 

分 类 号：R593.22[医药卫生—内科学]