芪丹通脉片下调NLRP3炎症小体对巨噬细胞泡沫化进程的影响  

The effect of Qidan Tongmai tablets on the foaming process of macrophages by down-regulating NLRP3 inflammasome

作　　者：王南丁 陈晋玉 李成龙 胡乔斌 雷瑗琳 张莎 李哲 WANG Nanding;CHEN Jinyu;LI Chenglong;HU Qiaobin;LEI Yuanlin;ZHANG Sha;LI Zhe(The Cardiovascular Department of Xi’an Traditional Chinese Medicine Hospital,Xi’an 710021,China)

机构地区：[1]西安市中医医院心血管内科,710021 [2]陕西中医药大学 [3]陕西中医药大学基础医学院 [4]陕西中医药大学第一临床医学院

出　　处：《环球中医药》2022年第12期2362-2367,共6页Global Traditional Chinese Medicine

基　　金：陕西省自然科学基础研究计划(2020JQ-966);西安市科协青年人才托举计划(095920201330);陕西中医药大学学科创新团队(2019-YS01);2021年西安市科技局项目(21YXYJ2256)。

摘　　要：目的分别观察益气中药、活血中药、益气活血配伍中药芪丹通脉片对氧化低密度脂蛋白(oxidized low densitylipoprotein,ox-LDL)诱导的小鼠单核巨噬细胞白血病细胞(RAW264.7细胞)泡沫化进程影响及其对泡沫化过程中核苷酸结合寡聚化结构域样受体蛋白3(NLR pyrin domain containing 3,NLRP3)炎症小体的影响。方法培养后的RAW264.7细胞,随机分正常组、ox-LDL诱导组益气组、活血组、复方组及复方+siRNA组,泡沫化诱导经中药干预后的各组细胞,分析其油红染色阳性面积,计算泡沫化诱导率,收集细胞,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)法检测白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)等炎性因子的表达水平,分别用实时荧光定量PCR(real-time PCR)及免疫印迹法(western blot,WB)分析手段分析NLRP3、半胱氨酸蛋白水解酶-1(Caspase-1)的表达水平。结果(1)对照组细胞几乎没有油红O着色,诱导后的各组细胞均出现不同程度的油红O着色,ox-LDL诱导组细胞油红着色面积比值最高,其余四组油红O着色面积比值与ox-LDL诱导组比较均降低均具有统计学意义,其中复方组及复方+siRNA组油红着色面积比值最低,与益气组及活血组比较均具有统计学意义。(2)对照组中IL-1β及IL-18含量最低,与对照组比较其它各组的IL-1β及IL-18含量均升高,益气组、活血组、复方组、复方+siRNA组与ox-LDL诱导组比较IL-1β及IL-18含量降低具有统计学意义,其中复方组与复方+siRNA组IL-1β及IL-18含量较益气组、活血组降低具有统计学意义。(3)对照组的NLRP3、Caspase-1含量最低;ox-LDL诱导组NLRP3、Caspase-1含量最高;药物干预的四组与ox-LDL诱导组相比,两个指标降低均具有统计学意义,其中复方组NLRP3、Caspase-1含量较益气组、活血组降低具有统计学意义。(4)ox-LDL诱导组NLRP3、Caspase-1含量最高,与ox-LDL诱导组比较,其它各组的NObjective To observe the effects of Qidantongmai tablet on the transformation process of RAW264.7 cells to the foam cells induced by ox-LDL and NLRP3 inflammasomes.Methods RAW264.7 cells were cultured and randomly divided into control group,ox-LDL induction group and ox-LDL induction+Yiqi group,ox-LDL induction+huoxue group,ox-LDL induction+QDTM group,ox-LDL induction+QDTM group+siRNA group.The area of Oil red positive express on each group was analyzed and the ratio of Oil red O positive area/cell total area was computed after Macrophage-to-foam Cell transformation.After collecting the cells,inflammatory factors IL-1β、IL18 were detected by ELISA,the levels of NLRP3 and caspase-1 were detected by Real-time PCR and Western blot.Results(1)The normal group cells almost no oil red O staining,groups of cells in different degrees after induction of oil red O staining,ox-LDL induced cell groups oil red staining area ratio is the highest,the rest of the four groups of oil red O staining area ratio is lower and ox-LDL induced group were statistically significant,the compound group and complex the ratio of oil red staining area in fang+siRNA group was the lowest,which was statistically significant compared with the Yiqi group and the blood activating group.(2)The contents of IL-1βand IL-18 in the control group were the lowest,and the contents of IL-1βand IL-18 in other groups were increased compared with the control group,the Yiqi group,Huoxue group and the compound group.The levels of IL-1βand IL-18 in the compound+siRNA group were significantly decreased than those in the ox-LDL-induced group,and the levels of IL-1βand IL-18 in the compound+siRNA group and the compound+siRNA group were significantly lower than those in the Yiqi group and the Huoxue group.(3)The content of NLRP3 caspase-1 in the control group was the lowest.The content of NLRP3 caspase-1 in the ox-LDL induced group was the highest.Compared with the ox-LDL-induced group,the two indexes in the four groups of drug intervention were significantly decre

关 键 词：益气活血 核苷酸结合寡聚化结构域样受体蛋白3炎症小体 芪丹通脉片 泡沫化 炎症反应 动脉硬化 中药复方 巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]