Yoon Hyun Joong, Moon Myoung E, Park Haeng Soon, Im Suhn Young, Kim Young Ho
College of Pharmacy, and Research Institute of Drug Development, Chonnam National University, Gwangju, Republic of Korea.
Biochem Biophys Res Commun. 2007 Jul 6;358(3):954-9. doi: 10.1016/j.bbrc.2007.05.042. Epub 2007 May 14.
The focus of this study was to clarify the relation between the nitric oxide (NO) production and cytokine expression including tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and also investigated the effect of COS on LPS stimuli from RAW 264.7 cell. The lipopolysaccharide (LPS) of Gram-negative bacteria induces the expression of cytokines and potent inducers of inflammatory cytokines such as TNF-alpha and IL-6. In this experiment, upon stimulation with increasing concentrations of chitosan, the LPS-stimulated TNF-alpha and IL-6 secretion was significantly recovered within the incubation media of RAW 264.7 cells. Consistently, RT-PCR with mRNA and Western blot with anti-cytokine antiserum including TNF-alpha and IL-6 showed that the amount of TNF-alpha and IL-6 secretion in the incubation media recovered with the concentration of chitosan. The LPS-stimulated NO secretion was significantly recovered within the 6h and 12h incubation media of RAW 264.7 cells, too. The recovery effect of chitosan on IL-6 and NO secretion may be induced via the stimulus of TNF-alpha in RAW 264.7 cell. These results once again suggest that chitosan oligosaccharide may have the anti-inflammatory effect via the stimulus of TNF-alpha in the LPS-stimulated inflammation in RAW 264.7 cells.
本研究的重点是阐明一氧化氮（NO）生成与包括肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）在内的细胞因子表达之间的关系，并研究壳寡糖对RAW 264.7细胞脂多糖（LPS）刺激的影响。革兰氏阴性菌的脂多糖（LPS）可诱导细胞因子以及TNF-α和IL-6等炎性细胞因子强效诱导剂的表达。在本实验中，随着壳聚糖浓度增加进行刺激时，RAW 264.7细胞孵育培养基中LPS刺激的TNF-α和IL-6分泌显著恢复。同样，对mRNA进行逆转录聚合酶链反应（RT-PCR）以及使用包括TNF-α和IL-6在内的抗细胞因子抗血清进行蛋白质免疫印迹分析表明，孵育培养基中TNF-α和IL-6的分泌量随壳聚糖浓度而恢复。RAW 264.7细胞孵育6小时和12小时的培养基中，LPS刺激的NO分泌也显著恢复。壳聚糖对IL-6和NO分泌的恢复作用可能是通过RAW 264.7细胞中TNF-α的刺激诱导的。这些结果再次表明，壳寡糖可能通过在RAW 264.7细胞的LPS刺激炎症中刺激TNF-α而具有抗炎作用。
