脉络宁口服液改善动脉硬化性闭塞症的体外药效与机制  

Effects and mechanism of Mailuoning oral liquid on arteriosclerosis occlusion in vitro

作　　者：来兴兆 范玲玲 黄素清 黄聪聪 李剑 谭宁华[1] LAI Xingzhao;FAN Lingling;HUANG Suqing;HUANG Congcong;LI Jian;TAN Ninghua(School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198;Jinling Pharmaceutical Co.,Ltd.,Nanjing 210009,China)

机构地区：[1]中国药科大学中药学院,南京211198 [2]金陵药业股份有限公司,南京210009

出　　处：《中国药科大学学报》2025年第1期91-98,共8页Journal of China Pharmaceutical University

摘　　要：以脂多糖(LPS)诱导小鼠单核巨噬细胞(RAW264.7)建立体外炎症模型,探究脉络宁口服液改善动脉硬化性闭塞症(arteriosclerosis occlusion,ASO)的体外药效与机制。通过MTT法检测RAW264.7细胞活力;Griess试剂法检测一氧化氮(nitric oxide,NO)的浓度;Q-PCR法和Western blot法检测NFAT5/NLRP3信号通路的mRNA与蛋白表达水平;细胞转染NFAT5-siRNA联合Western blot法探究NFAT5与NLRP3的关系;免疫荧光法检测NFAT5核转位。结果显示:脉络宁口服液减少LPS诱导的RAW264.7细胞NO释放,下调NFAT5、NLRP3、caspase1、IL-18和MMP9的mRNA水平,抑制NFAT5、NLRP3、cleaved-caspase1(p20)的蛋白表达量以及NF-κB-P65磷酸化水平;NFAT5-siRNA显著逆转LPS诱导的RAW264.7细胞中NLRP3的蛋白表达增加;脉络宁口服液与KRN2(NFAT5抑制剂)均能够抑制NFAT5表达与核转位。综上所述,脉络宁口服液在体外通过抑制NFAT5/NLRP3信号通路发挥显著抗炎作用,并且NFAT5可能参与调控NLRP3的表达。Lipopolysaccharide(LPS)was used to induce mouse mononuclear macrophages(RAW 264.7 cells)to establish the inflammation model for investigating the effect and mechanism of Mailuoning oral liquid on arteriosclerosis occlusion(ASO)in vitro.RAW264.7 cells viability was measured by MTT assay.NO concentration was determined by Griess.mRNA levels and protein expressions of NFAT5/NLRP3 signaling pathway were detected by Q-PCR and Western blot.The relationship between NFAT5 and NLRP3 was explored by cellular transfection of NFAT5-siRNA combined with Western blot.Nuclear translocation of NFAT5 was detected by immunofluorescence.The results showed that Mailuoning oral liquid decreased the NO release induced by LPS in RAW264.7 cells.The mRNA levels of NFAT5,NLRP3,caspase1,IL-18 and MMP9,the protein expressions of NFAT5,NLRP3,cleaved-caspase1(p20)and the phosphorylation of NF-κB-P65 were decreased after administration of Mailuoning oral liquid.NFAT5-siRNA significantly reversed the increase in protein expressions of NLRP3 induced by LPS in RAW264.7 cells.Both Mailuoning oral liquid and KRN2(NFAT5 inhibitor)could inhibit the expressions and nuclear translocation of NFAT5.In conclusion,Mailuoning oral liquid exert significant anti-inflammatory effects in vitro by inhibiting the NFAT5/NLRP3 signaling pathway,and NFAT5 might be involved in regulating the expressions of NLRP3.

关 键 词：动脉硬化性闭塞症 脉络宁口服液 RAW264.7巨噬细胞 NFAT5/NLRP3信号通路 炎症 

分 类 号：R285.5[医药卫生—中药学]