LPS介导三种啮齿动物原代胶质细胞炎症氧化反应的研究
A Study of LPS Mediated Inflammatory Oxidation Reaction of Three Types of Rodents
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摘要: 目的 以Sprague-Dawley大鼠 (SD Rats) 、C57BL/6小鼠和昆明小鼠 (Kunming mice) 为研究对象, 探索研究脂多糖 (Lipopolysaccharide, LPS) 诱导3种鼠的原代混合胶质细胞释放炎症因子的对比.从而进一步探讨、发现研究三种鼠的应用价值、寻找更理想的炎症模型.方法 以一氧化氮合酶 (i NOS) 、环氧化酶 (COX-2) 、白介素-1beta (IL-1β) 和Nitric Oxide (NO) 作为代表激活的炎症氧化应激反应表征.获得C57BL/6小鼠、昆明种小鼠和SD大鼠23 d龄乳鼠后, 进行断头取脑和胶质细胞培养.传代至第3代接种于24孔板使用, 经LPS介导24 h后分别用Griess法检测NO浓度变化, Western b Lot检测COX-2、IL-1β、i NOS的蛋白表达.结果 用含N2的无血清培养基处理后的SD大鼠混合胶质细胞形态发生改变.比较三种鼠混合胶质细胞的正常对照组和LPS组, LPS组的NO释放量显著升高 (P<0.01) ;并且3种鼠混合胶质细胞的LPS组的i NOS/COX-2/IL-1β的表达也明显升高.结论 LPS可以诱导C57BL/6小鼠、昆明鼠以及SD大鼠混合胶质细胞NO的释放以及IL-1β、i NOS、COX-2蛋白的表达, 从而诱发炎症反应;LPS可以诱导3种鼠的原代混合胶质细胞形成炎症氧化模型.并且, SD大鼠反应较为灵敏.Abstract: Objective By comparing with the SD rats mixed glial cells, C57 mice mixed glial cells and Kunming mice mixed glial cells and exploring the expression of inflammatory factor of three mixed glial cells induced by lipopolysaccharide (LPS) , the study aimed to explore the application value of three kinds of mice and find the ideal model of inflammation. Me thods We used LPS as inducers, and NO, IL-1β, COX-2 and i NOS as anti-inflammatory antioxidant index. After cutting the head and taking the brain, we cultured the mixed glials. Then we used Greiss assay to detect the expression of NO and used western blot to detect the expression of protein of IL-1β, COX-2, and i NOS protein.Finally we compared the mixed glial cells from SD rats, C57 mice mixed glial cells and Kunming mouse mixed glial cells and selected the best inflammation model from three mixed glial cells.Re s ults The results showed that the morphological changes of the mixed glial cells in SD rats were treated with N2-free medium. Compared with the control group, the quantity of NO of LPS group of three mixed glial cells increased significantly (P<0.01) . The LPS group of SD rats released the highest concentration of NO. Western blot was used to detect the expression of IL-1β, Cox-2 and i NOS in three kinds of rodents. Compared with the blank control group, the expression of COX-2 protein, i NOS and IL-1β in the LPS group of the three mice increased significantly. The results showed that LPS could successfully stimulate the release of inflammatory cytokines in three kinds of mice, among which the SD rats were more sensitive and it could be used in the study of AD inflammation model. Conclus ion The results showed that LPS could induce the release of NO and the expression of IL-1β, i NOS and COX-2 in C57 BL/6 mice, Kunming mice and SD rats to induce inflammatory response. Thus, LPS can induce the formation of inflammatory oxidation models of the original mixed glial cells of the three mice. Moreover, the SD rats were more sensitive.
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Key words:
- LPS /
- Kunming Mouse /
- C57 Mouse /
- SD rats /
- NO /
- Interleukin-1β
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