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首页> 外文期刊>Cytokine >The effect of minocycline on indolamine 2, 3 dioxygenase expression and the levels of kynurenic acid and quinolinic acid in LPS-activated primary rat microglia
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The effect of minocycline on indolamine 2, 3 dioxygenase expression and the levels of kynurenic acid and quinolinic acid in LPS-activated primary rat microglia

机译：米诺环素对吲哚胺2,3中二恶英酶表达和LPS活化原代大鼠小鼠微胶质细胞蛋白酸和喹啉酸水平的影响

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Microglia are one of the most important neural cells in the central nervous system (CNS) which account for 10-15% of all cells found in the brain. A vast majority of studies indicate that microglia play a pivotal role in protection and damage of the CNS. It has been shown that microglia are mainly scavenger cells but also produce a barrage of factors that are involved in tissue repair and neural regeneration. Several lines of evidence indicate that unregulated activation of microglia in response to either endogenous or exogenous insults results in the production of toxic factors that propagate neuronal injury. Studies demonstrated that the activated microglia secret the excessive amounts of quinolinic acid (QA) and kynurenic acid (KYNA) which are highly toxic for the neuronal cells. In line with this, indolamine 2, 3 dioxygenase (IDO), an enzyme producing KYNA and QA has been shown to be elevated during the inflammation in microglia. In this study, we established primary microglial cell cultures obtained from cerebral cortices of 1-day neonatal Wistar rats. Minocycline (20-60 mu M) or its vehicle was added to the culture media 60 min prior to 48 h incubation with lipopolysaccharide (LPS; 10 ng/mL). Using a specific process of adhesion and shaking of the cultured glial cells, a purified culture of approximately 94% enriched microglia was obtained and then, corroborated by immunocytochemistry (ICC). The cell viability after minocycline treatments was assessed using the MTT colorimetric assay. The expression of IDO was evaluated using qPCR. The levels of KYNA and QA were determined using enzyme-linked immunosorbent assay (ELISA). The results showed that minocycline significantly decreased the levels of both KYNA and QA in glia cells exposed to LPS. Moreover, minocycline decreased the expression of IDO in treated LPS-induced microglia. It seems that minocycline has a potent ability to oppress the inflammatory process via the decrease in production of IDO expression and the concentrations of KYNA and QA.
机译：微胶鸡是中枢神经系统（CNS）中最重要的神经细胞之一，其占大脑中发现的所有细胞的10-15％。绝大多数研究表明，微胶质细胞在CNS的保护和损害中发挥着关键作用。已经表明，小胶质细胞主要是清除剂细胞，但也产生了参与组织修复和神经再生的因素的阻碍。几种证据表明，响应于内源或外源性损伤的引起的微胶质引起的引入导致繁殖神经元损伤的毒性因素的产生。研究表明，活化的微胶质细胞批定过量的喹啉酸（QA）和蛋白质酸（Kyna），其对神经元细胞具有高度毒性。符合此，在微胶质细胞的炎症期间，吲哚胺2,3二氧化根酶（IDO），产生Kyna和QA的酶。在这项研究中，我们建立了从1天新生儿Wistar大鼠的脑皮质获得的初级小胶质细胞培养物。在与脂多糖（LPS; 10ng / ml）孵育之前60分钟将米诺环素（20-60μm）或其载体加入到培养基中。使用培养的胶质细胞的粘附和摇动的特定方法，得到约94％的富集的微胶质细胞的纯化培养，然后通过免疫细胞化学（ICC）来证实。使用MTT比色测定评估米诺环素治疗后的细胞活力。使用QPCR评估IDO的表达。使用酶联免疫吸附测定（ELISA）测定Kyna和QA的水平。结果表明，米诺霉素在暴露于LPS暴露于LPS的胶质细胞中的Kyna和QA的水平显着降低。此外，米诺环素减少了IDO治疗LPS诱导的小胶质细胞的表达。似乎米诺环素似乎通过IDO表达的减少和kyna和Qa的浓度降低了炎症过程的能力。

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