摘要
目的探讨黄蜀葵花中5种黄酮类化合物对肠道L细胞AGEs/RAGE/p38MAPK/NF-κB信号通路的调节作用。方法200 mg·L^(-1)AGEs作用肠道于L细胞株GLUTag细胞,Western blot检测细胞中RAGE、p22^(Phox)、p47^(Phox)、p53、Bax蛋白的相对表达量,ELISA法检测TNF-α、IL-1、IL-6、caspase-3、caspase-9的含量。实验分为7组:空白对照组(NG)、模型组(AGEs)、槲皮素组(QT)、异槲皮苷组(IQT)、金丝桃苷组(HY)、槲皮素-3'-O-葡萄糖苷组(QG)、棉皮素-8-O-葡萄糖醛酸苷组(GG)。NG细胞培养于DMEM低糖培养基中;AGEs细胞培养于含200 mg·L^(-1)AGEs的DMEM低糖培养基中;给药组细胞分别培养于含50μmol·L^(-1)各黄酮单体及含200 mg·L^(-1)AGEs的DMEM低糖培养基中。各组细胞经药物干预24 h后,Western blot检测细胞中RAGE、p22^(Phox)、p47^(Phox)、p53、Bax、p-p38MAPK、NF-κB蛋白的相对表达量,ELISA法检测TNF-α、IL-1、IL-6的含量及caspase-3、caspase-9的活性。结果模型组细胞中RAGE、p22^(Phox)、p47^(Phox)、Bax、caspase-3、caspase-9、Phospho-p38MAPK、NF-κB相对表达量及细胞上清液中TNF-α、IL-1、IL-6的分泌量明显上调(P<0.01),p53相对表达量明显下调(P<0.01)。50μmol·L^(-1)槲皮素、异槲皮苷、金丝桃苷、槲皮素-3'-O-葡萄糖苷、棉皮素-8-O-葡萄糖醛酸苷分别给药后,各蛋白的相对表达量及炎症因子的分泌量接近正常组。结论黄属葵花中5种黄酮类化合物能明显抑制肠道L细胞AGEs/RAGE/p38MAPK/NF-κB信号通路。
Aim To investigate the regulation of five flavonoids from Abelmoschl manihot on the AGEs/RAGE/p38MAPK/NF-κB signaling pathway in GLUTag cells. Methods With GLUTag cells treated with 200 mg · L^(-1)AGEs,Western blot analysis was applied to determine the expression of RAGE,p22 ^(Phox),p47^(Phox),p53 and Bax proteins and ELISA analysis was used to detect the levels of TNF-α, IL-1, IL-6,caspase-3,caspase-9. The cells were divided into seven groups: the normal group( NG), model group( AGEs),quercetin-treated group( QT),isoquercitrintreated group( IQT),hyperoside-treated group( HY),quercetin-3'-O-glucoside-treated group( QG),gossypetin-8-O-glucuronide-treated group( GG). NG cells were cultured in DMEM low-glucose culture medium without adding any other intervention factors; model group cells were cultured in DMEM low-glucose medium containing 200 mg·L^(-1)AGEs; cells in five treatment groups were cultured in DMEM low-glucose culture medium containing 200 mg · L^(-1)AGEs,which were respectively added 50 μmol·L^(-1)QT,IQT,HY,QG,GG. Western blot analysis was applied to determine the expression of RAGE,p22 ^(Phox),p47^(Phox),p53 and bax proteins and ELISA analysis was used to detect the levels of TNF-α,IL-1,IL-6,caspase-3,caspase-9. Results The contents of RAGE,p22 ^(Phox),p47^(Phox),bax,caspase-3,caspase-9,phospho-p38 MAPK,NF-κB and TNF-α,IL-1 and IL-6 significantly increased in model group cells( P < 0. 01),and the relative expression of p53 was significantly down-regulated( P <0. 01) in model group cells. After treated with 50μmol · L^(-1)quercetin, isoquercitrin, hyperoside,quercetin-3'-O-glucoside,gossypetin-8-O-glucuronide,the content of proteins and inflammatory factors were close to normal group. Conclusion The five flavonoids in Abelmoschus manihot( L.) Medicus can inhibit AGEs/RAGE/p38MAPK/NF-κB signaling pathway in GLUTag cells.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2017年第6期798-805,共8页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81473408,81673533)
江苏高等学校中药学优势学科建设项目(No ysxk-2014)
教育部新世纪优秀人才计划项目(No NCET-13-0873)
