老瓜头生物总碱对LPS诱导RAW264.7细胞中COX-2酶活性、mRNA及蛋白表达的影响  

Effect of total alkaloids of Cynanchum Komarovii AI. Icjinski on COX-2 enzyme activity and COX-2 mRNA /protein expression in LPS-stimulated RAW264. 7 cells

作　　者：杨琤琤 陈亚寒 杨文博[3] 冉林武[3] 杨卫东[1,3] 

机构地区：[1]宁夏医科大学药学院,银川750004 [2]银川市第一人民医院,银川750004 [3]宁夏医科大学实验动物中心,银川750004

出　　处：《中国新药杂志》2014年第9期1081-1085,共5页Chinese Journal of New Drugs

基　　金：宁夏回族自治区自然科学基金(NZ11107)

摘　　要：目的:研究老瓜头生物总碱(TACKI)对脂多糖(LPS)诱导小鼠巨噬细胞RAW264.7在COX-2酶活性、基因表达及蛋白表达的影响。方法:采用LPS诱导RAW264.7细胞作为炎症模型,以非甾体抗炎药选择性COX-2酶抑制剂塞来昔布作为阳性对照。MTT比色法分析TACKI对RAW264.7生长活性的影响,LPS长时间刺激RAW264.7,ELISA检测细胞上清液中前列腺素-E2(PGE2)活性,以PGE2的活性反映COX-2的酶活性,半定量RT-PCR检测RAW264.7细胞中COX-2 mRNA的表达,免疫荧光观察RAW264.7细胞中COX-2的表达,Western blotting法检测COX-2蛋白表达。结果:与LPS组比较,TACKI 0.1,1μg·L-1能减弱COX-2酶活性(P<0.05),TACKI 0.01,0.1,1μg·L-1可下调COX-2 mRNA的表达(P<0.05),TACKI 0.1,1μg·L-1能减少COX-2蛋白表达(P<0.05)。结论:TACKI减弱LPS诱导RAW264.7细胞COX-2酶活性,对LPS诱导RAW264.7细胞中COX-2基因表达及蛋白表达抑制作用明显。Objective ： To assess the effect of total alkaloids of Cynanchum Komarovii AI. Icjinski （ TAC- KI） on the COX-2 enzyme activity and COX-2 expression in LPS-stimulatd RAW264.7 cells. Methods： The effect of TACKI on RAW264.7 cells was determined by MTT assay. After pretreated with various concentrations of TAC- KI for 2 h, the cells were exposed to 1 mg. L^-1 LPS for 22 h. The activity of COX-2 enzyme in RAW264.7 cells was analyzed by ELISA. The expressions of COX-2 mRNA and protein were analyzed by RT-PCR and Western blot- ting, and the expression of COX-2 protein was also determined by immunofluorescence. Results： TACKI inhibited COX-2 enzyme activity at 0.1 and 1 μg. L^-1, significantly inhibited COX-2 mRNA expression at 0.01, 0. 1 and 1 μg.L^-1, and inhibited COX-2 protein expression at 0. 1 and 1 μg.L^-1. Conclusion： TACKI has an inhibitory effect on COX-2 enzyme activity and the expression of COX-2 mRNA and protein in LPS-stimulated RAW264.7 cells.

关 键 词：老瓜头生物总碱 环氧合酶-2 脂多糖 抗炎 

分 类 号：R961.5[医药卫生—药理学]