摘要
目的:以短穗兔耳草中分离得到的苯乙醇苷类代表化合物大车前苷及黄酮苷元的代表化合物苜蓿素和柯伊利素为研究对象,研究其对尿酸钠结晶(MSU)诱导的RAW264.7巨噬细胞炎症模型的影响及作用机理。方法:用MSU诱导RAW264.7小鼠单核巨噬细胞建立炎症模型,分为空白对照组、模型对照组、秋水仙碱0.001μmol/L组、大车前苷0.078、0.156μmol/L组、苜蓿素0.078、0.156μmol/L组;柯伊利素0.031、0.062μmol/L组。在药物干预12 h后酶联免疫吸附测定法检测上清液中的肿瘤坏死因子-α(TNF-α)含量;Western blot法检测全细胞提取液中核转录因子κB(NF-κB)、Toll样受体4(TLR4)、TLR2、半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)和NOD样受体蛋白3(NLRP3)的蛋白表达。结果:与空白对照组相比,模型对照组TNF-α含量及NF-κB、TLR4、TLR2、Caspase-1、NLRP3蛋白表达显著上调(P<0.01);与模型对照组相比,秋水仙碱0.001μmol/L组与3种化合物均能显著降低TNF-α含量(P<0.01),大车前苷0.078μmol/L组TLR4、NLRP3蛋白表达显著下调(P<0.01),大车前苷0.156μmol/L组TLR4、Caspase-1、NLRP3蛋白表达显著下调(P<0.01),苜蓿素0.078μmol/L组TLR4蛋白表达显著下调(P<0.01),苜蓿素0.156μmol/L组NF-κB、TLR4、Caspase-1、NLRP3蛋白表达显著下调(P<0.01),柯伊利素0.031、0.062μmol/L组NF-κB、TLR4、TLR2、Caspase-1、NLRP3蛋白表达显著下调(P<0.01)。结论:大车前苷、苜蓿素和柯伊利素可能通过调控NF-κB、TLR4、TLR2、Caspase-1及NLRP3的表达抑制RAW264.7巨噬细胞炎症模型TNF-α炎症因子的释放进而发挥抗炎作用。
Objective:To investigate the effect of phenylethanoid glycoside(plantamajoside)and flavonoid aglycone(tricin and chrysoeriol)on the inflammation model of RAW264.7 macrophages induced by sodium urate crystals(MSU),and to explore its mechanism.Methods:MSU was used to induce inflammation in mouse mononuclear macrophages RAW264.7.The mice were divided into blank control group,model control group,colchicine(0.001μmol/L)group,plantamajoside(0.078 and 0.156μmol/L)groups,tricin(0.078 and 0.156μmol/L)groups,and chrysoeriol(0.031 and 0.062μmol/L)groups.After 12 h of drug intervention,the content of tumor necrosis factorα(TNF-α)in the supernatant was detected by enzyme-linked immunosorbent assay(ELISA),and the protein levels of nuclear factorκB(NF-κB),Toll-like receptor 4(TLR4),TLR2,caspase 1,and NOD-like receptor protein 3(NLRP3)in the whole cell extract were detected by Western blot.Results:Compared with the blank control group,the model control group showed increased TNF-αcontent and up-regulated protein levels of NF-κB,TLR4,TLR2,Caspase-1,and NLRP3(P<0.01).As compared with the model control group,the content of TNF-αwas decreased in the colchicine 0.001μmol/L group and the plantamajoside,tricin,and chrysoeriol groups(P<0.01).The protein levels of TLR4 and NLRP3 were down-regulated in the plantamajoside 0.078μmol/L group(P<0.01),and the protein levels of TLR4,Caspase-1,and NLRP3 were down-regulated in the plantamajoside 0.156μmol/L group(P<0.01).The protein level of TLR4 was decreased in the tricin 0.078μmol/L group(P<0.01),and the protein levels of NF-κB,TLR4,Caspase-1,and NLRP3 were reduced in the tricin 0.156μmol/L group(P<0.01).The chrysoeriol(0.031 and 0.062μmol/L)groups exhibited down-regulated protein levels of NF-κB,TLR4,TLR2,Caspase-1,and NLRP3(P<0.01).Conclusion:Plantamajoside,tricin,and chrysoeriol may have anti-inflammatory effects by inhibiting the release of TNF-αinflammatory factor in RAW264.7 macrophage inflammation model through regulating the expressions of NF-κB,TLR4,TLR2,Caspase-1
作者
郭敏侠
欧阳香
王陆
周睿卿
黄小方
程虹毓
朱继孝
Guo Minxia;Ouyang Xiang;Wang Lu;Zhou Ruiqing;Huang Xiaofang;Cheng Hongyu;Zhu Jixiao(Research Center of Chinese Medicine Resources and Ethnic Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004;Chia Tai Tianqing Pharmaceutical Group Co.,Ltd.,Nanjing 211100;College of Traditional Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004;School of Humanities,Jiangxi University of Chinese Medicine,Nanchang 330004)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第2期49-53,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金项目(编号:82060757)
江西省教育厅科学技术研究项目重点项目(编号:GJJ190632)
国家重点研发计划项目(编号:2019YFC1712302)
