宁心涤痰汤通过AMPK/NF-κB通路抑制巨噬细胞分化改善血管再狭窄的机制研究  

Mechanism of Ningxin Ditan Decoction in Inhibiting In-stent Restenosis by AMPK/NF-kB Pathway Mediated Macrophage Differentiation

作　　者：曹丽娟[1,2] 于玲 刘莉 Cao Lijuan;Yu Ling;Liu Li(Heilongjiang University of Chinese Medicine,Heilongjiang,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江省齐齐哈尔市中医医院,黑龙江齐齐哈尔161099 [3]广东省中医院,广东广州510120 [4]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《中国中医急症》2023年第4期592-596,623,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82104962);黑龙江省青年人才托举工程项目(2022-QNRC1-28)。

摘　　要：目的 研究宁心涤痰汤通过AMPK/NF-κB通路抑制巨噬细胞分化改善经皮冠状动脉介入术(PCI)后血管再狭窄的分子机制。方法 将人急性单核细胞白血病细胞(THP-1)体外诱导分化成巨噬细胞,并分为空白组、模型组、宁心涤痰汤低、中、高剂量组和辛伐他汀组。除空白组外,余下各组通过氧化修饰型低密度脂蛋白(ox-LDL)诱导构建泡沫细胞模型,并给予相应的含药血清进行干预。分别检测各组细胞内胆固醇的含量,采用酶联免疫吸附试验(ELISA)检测细胞上清中炎症因子[白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)]的含量,流式细胞术检测巨噬细胞表型(CD11b+CD86+)及凋亡的影响,Western blotting检测细胞中AMPK/NF-κB通路中相关蛋白的表达水平。结果 与空白组比较,模型组中游离胆固醇(FC)、总胆固醇(TC)、IL-6、IL-1β、TNF-α的含量升高和CD11b+CD86+及凋亡的比例显著升高,细胞内p-AMPK及细胞质NF-κB p65蛋白的表达显著降低,细胞核NF-κB p65蛋白的表达显著上调,差异均有统计学意义(P <0.01);与模型组比较,宁心涤痰汤各剂量及辛伐他汀均可显著抑制细胞内TC、FC、IL-6、IL-1β、TNF-α的含量和CD11b+CD86+及凋亡的比例,上调细胞内p-AMPK及细胞质NF-κB p65蛋白的表达,下调细胞核NF-κB p65蛋白的表达水平,差异均有统计学意义(P <0.05或P <0.01)。结论 宁心涤痰汤通过抑制AMPK/NF-κB信号通路来抑制巨噬细胞分化,达到预防PCI术后血管再狭窄的目的。Objective:To observe the effect of Ningxin Ditan Decoction on AMPK/NF-kB pathway mediated macrophage difierentiation and to analyze its molecular mechanism of improving in-stent restenosis after percutaneous coronary intervention.Methods:THP-1 was induced to differentiate into macrophages in vitro,and were divided into control group,model group,Ningxin Ditan Decoction low-,middle-and high-dose groups,and simvastatin group.In addition to the control group,the remaining groups were induced by oxidized low-density lipoprotein(ox-LDL)to build a foam cell model,and the corresponding drug containing serum was given for intervention.The content of cholesterol in cells of each group was detected,and the inflammatory factors(IL-6,TNF-a and IL-1P)in the supernatant of cells were detected by enzyme-linked immunosorbent assay(ELISA).The expression of macrophage phenotype(CD 11 b+CD86+)and apoptosis were detected by flow cytometry,and the expression level of related proteins in AMPKZNF-kB pathway were detected by Western blotting.Results:Compared with the control group,the content of TC,FC,IL-6,TNF-a and IL-lp and the ratio of CDllb+CD86+and apoptosis of macrophages were significantly increased,p-AMPK and cytoplasmic NF-kB p65 protein was significantly reduced,while the expression of nuclear NF-kB p65 protein was significantly up-regulated in the model group,and the difference was statistically significant(P<0.01).Compared with the model group,Ningxin Ditan Decoction and simvastatin could significantly inhibit intracellular TC,FC,IL-6,IL-1 p,TNF-a contents and the ratio of CDllb+CD86+and apoptosis,up-regulated p-AMPK and cytoplasmic NF-kB p65 protein expression,down regulating the expression level of nuclear NF-kB p65,and the difiference was statistically significant(P<0.05 and P<0.01).Conclusion:Ningxin Ditan Decoction inhibits the differentiation of macrophages through inhibiting AMPK/NF-kB signal pathway to prevent in-stent restenosis after percutaneous coronary intervention.

关 键 词：血管再狭窄 宁心涤痰汤 巨噬细胞 AMPK/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]