孤儿核受体Nur77对氧化型低密度脂蛋白诱导的小鼠巨噬细胞系分泌炎症因子的影响  

Effect of orphan nuclear receptor Nur77 on inflammatory factor secretion in murine macrophage strain

作　　者：胡刘华[1] 何奔[1] 沈玲红[1] 江立生[1] 柴大军[1] 王力[2] 曾锦章[2] 

机构地区：[1]上海交通大学医学院附属仁济医院心内科,上海200127 [2]中科院上海生命科学研究院生茂联合研发中心,上海200233

出　　处：《心脏杂志》2008年第5期541-544,共4页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(30670880)

摘　　要：目的观察孤儿核受体Nur77对氧化型低密度脂蛋白(ox-LDL)诱导小鼠巨噬细胞分泌炎症因子的影响。方法建立稳定转染GFP-Nur77质粒的小鼠巨噬细胞系RAW 264.7,并以稳定转染绿色荧光蛋白(GFP)基因的细胞系RAW 264.7作为对照。将转染的细胞与ox-LDL(40 mg/L)共同孵育24 h后,采用ELISA法检测炎症因子TNF-α、IFNγ-、基质金属蛋白酶-9(MMP-9)、单核细胞趋化蛋白-1(MCP-1)和IL-6的表达。结果经ox-LDL刺激后,不同质粒DNA转染的两种细胞中,IFNγ-、TNF-α、MMP-9和MCP-1的浓度均升高(P<0.05);但GFP-Nur77-RAW 264.7细胞的增长水平明显低于GFP-RAW 264.7细胞(P<0.05)。ox-LDL刺激前,GFP-RAW 264.7细胞分泌的4种炎症因子(TNF-α、IFN-γ、MMP-9和MCP-1)的浓度,分别为(19.00±1.69)ng/L、(99.10±3.09)ng/L、(0.14±0.02)ng/L和(23.96±3.57)ng/L;ox-LDL刺激后,分别上升为(27.22±0.38)ng/L、(549.90±1.81)ng/L、(0.45±0.06)ng/L和(49.31±3.66)ng/L。ox-LDL刺激前,GFP-Nur77-RAW 264.7和GFP-RAW 264.7细胞分泌的4种炎症因子的浓度无统计学的差异性。ox-LDL刺激后,4种炎症因子的浓度分别上升为(18.34±1.06)ng/L、(320.56±1.27)ng/L、(0.20±0.03)ng/L和(38.92±4.46)ng/L。而两种细胞(CTFP-Nur77-RAW 264.7细胞与GFP-RAW 264.7细胞)分泌IL-6的浓度未见显著差异。结论RAW 264.7细胞中Nur77表达的上调,能够显著减少ox-LDL诱导的炎症因子(IL-6除外)分泌,这可能是孤儿核受体Nur77心血管保护作用的重要机制之一。AIM To observe the effect of orphan nuclear receptor Nur77 on inflammatory factor secretion in murine maerophage RAW264. 7 induced by oxidized low density lipoprotein （ox-LDL）. METHODS RAW264. 7 cells stably expressing transfecting plasmid GFP-Nur77 and GFP cDNA（ as a control） were respectively established. After the two transfeeted macrophages were incubated with ox-LDL （40 mg/L）, the concentrations of TNF-α, IFN-γ, MMP-9, MCP-1 and IL-6 in culture supernatant were detected by ELISA. RESULTS After the ox-LDL stimulation, the concentrations of IFN-γ, TNF-α, MMP-9 and MCP-1 in the two transfeeted macrophages all increased （P 〈0. 05）. However, the increased concentrations in GFP-Nur77-RAW264. 7 were significantly less than those in GFP-RAW264.7 （P 〈 0. 05 ）. Before ox-LDL stimulation, the concentrations of TNF-α, IFN-γ, MMP-9 and Mcp-1 in GFP-RAW264.7 were respectively （ 19. 00 ± 1.69） ng/L, （99. 10 ± 3.09 ） ng/L, （0. 14 ± 0. 02 ） ng/L and （23.96 ± 3.57） ng/L. After the stimulation, the concentrations of the 4 inflammatury factors increased to （27.22 ±0.38） ng/L, （549.90 ± 1.81） ng/L, （0.45 ±0.06） ng/L and （49. 31 ±3.66） ng/L respectively. Before ox-LDL stimulation, the concentrations of the 4 inflammatory factors secreted by GFP-Nur77- RAW264. 7 and GFP-RAW264.7 had no statistical difference. After ox-LDL stimuladtion, the concentrations of the 4 inflammatory factors increased to （ 18.34 ± 1.06） ng/L, （320.56 ± 1.27） rig/L, （0. 20 ± 0. 03） ng/L and （38.92 ± 4. 46） ng/L respectively. However, no significant changes were observed in IL-6 concentration secreted by GFP-Nur77-RAW264. 7 or GFP-RAW264.7. CONCLUSION High expression of Nur77 on RAW264.7 cells obviously reduce the inflammatory factor secretion induced by ox-LDL. This may be an important protective mechanism of Nur77.

关 键 词：孤儿核受体 氧化型低密度脂蛋白 巨噬细胞 炎症因子 小鼠 

分 类 号：R513.5[医药卫生—内科学]