云芝糖肽下调脂多糖诱导的小鼠巨噬细胞炎症性因子  

Down-regulating effects of Coriolus versicolor polysaccharopeptide on murine macrophage production of inflammatory mediators induced by lipopolysaccharide

作　　者：任文智[1] 杨晓彤[1,2] 马伟超[1] 冯慧琴[1] 杨明俊[1] 杨庆尧 董莎莎[1] 

机构地区：[1]上海师范大学微生物与免疫学研究所,上海200234 [2]上海芝草生物技术有限公司,上海200234

出　　处：《中国新药与临床杂志》2010年第4期306-309,共4页Chinese Journal of New Drugs and Clinical Remedies

基　　金：国家科技部创新基金项目(09C2613100756);上海市科委创新基金项目(0802H48400);上海市徐汇区科委创新基金(0A2006020)

摘　　要：目的探讨云芝糖肽(PSP)对脂多糖(LPS)诱导的炎症性因子产生的影响及其相关机制。方法小鼠单核巨噬细胞Raw 264.7常规培养,并分为模型组(加入1 mg·L-1 LPS)、PSP组(25 mg·L-1PSP预孵1 h后加入1 mg·L-1 LPS)和正常对照组。分组后细胞继续培养24 h,光镜观察各组细胞的形态变化,Griess试剂检测一氧化氮(NO)生成量,ELISA法检测花生四烯酸2(PGE2)和白细胞介素-1β(IL-1β)的含量,Western blotting方法检测诱导型一氧化氮合酶(iNOS)和选择性环氧合酶-2(COX-2)的表达,流式细胞术检测PSP对LPS-FITC与各组细胞结合后的荧光强度变化。结果光镜观察显示,模型组大量细胞呈过度激活形态,PSP组细胞的伪足和胞浆内颗粒物的明显较少。与模型组相比,PSP组的NO、PGE2和IL-1β含量显著减少(P<0.05或P<0.01),iNOS和COX-2的高表达显著降低(P<0.01)。PSP组结合到Raw 264.7细胞上的FITC-LPS量减少,细胞平均荧光强度从2.1±s 0.4下降到1.20±0.25。结论PSP能干扰LPS与巨噬细胞的结合,从而减少LPS诱导产生的炎症介质,具有一定的抗炎效果。AIM To study the effects and mechanisms of Coriolus versicolor polysaccharopeptide （PSP） on murine macrophage production of inflammatory mediators induced by lipopolysaccharide （LPS）. METHODSMurine macrophage cell line Raw 264.7 was routinely maintained, and stimulated with 1 mg.L^-1 LPS in modelgroup, pretreated with 25 mg ·L^-1 PSP for 1 h and then stimulated with 1 mg .L^-1 LPS in PSP group, respectively. Cells received culture medium only was served as control group. After further 24 h incubation, the morphological changes were observed under microscope. Nitric oxide （NO） production was detected with Griess＇s reagent. Prostaglandin E2 （PGE2） and interleukin-1β （IL-1β） were measured by ELISA method. Inducible nitric oxide synthase （iNOS） and cyclooxygenase-2 （COX-2） enzyme expressions were detected by Western blotting. Flow cytometry was applied to detect the effect of PSP on the binding of FITC-conjugated LPS to RAW 264.7. RESULTS Morphological observation showed that a large number of cells in model group were highly activated, but less pseudopods and intracytoplasmic granules were observed in PSP group. Compared with the model group, the productions of NO, PGE2, IL-1β and the high expressions of iNOS, COX2 were markedly decreased in the PSP group. The binding of FITC-conjugated LPS to the murine macrophage cell line RAW 264.7 were less in the PSP group, and consequently the mean fluorescent intensity decreased from 2.1 ± s 0.4 to 1.20 ± 0.25. CONCLUSION PSP could inhibit the binding of LPS to macrophage and decrease the high production of LPS stimulated inflammatory mediators.

关 键 词：云芝糖肽 脂多糖类 巨噬细胞 炎症 

分 类 号：R285.5[医药卫生—中药学]