论文部分内容阅读
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。
Objective To investigate the effects of PSP on the inflammatory factors induced by lipopolysaccharide (LPS) and its related mechanisms. Methods The RAW 264.7 cells were cultured in a routine manner and were divided into model group (1 mg · L -1 LPS) and PSP group (25 mg · L -1 PSP) LPS) and normal control group. The cells were further cultured for 24 hours after grouping. The morphological changes of the cells were observed under light microscope. The production of nitric oxide (NO) was detected by Griess reagent. The levels of arachidonic acid 2 (PGE2) and interleukin-1β (IL- 1β) were detected by Western blotting. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were detected by Western blotting. Changes in fluorescence intensity after binding. Results Light microscopy showed that a large number of cells in the model group showed an over-activated morphology. Pseudopodia and cytoplasmic particles in the PSP group were significantly less. Compared with model group, the levels of NO, PGE2 and IL-1βwere significantly decreased in PSP group (P <0.05 or P <0.01), while the expression of iNOS and COX-2 in PSP group was significantly decreased (P <0.01). The amount of FITC-LPS bound to RAW 264.7 cells in PSP group decreased, and the mean fluorescence intensity decreased from 2.1 ± 0.4 to 1.20 ± 0.25. Conclusion PSP can interfere with the binding of LPS to macrophages, thereby reducing the inflammatory mediators induced by LPS, and has some anti-inflammatory effects.