Responses of rat and mouse primary microglia to pro- and anti-inflammatory stimuli: molecular profiles, K(+) channels and migration.
Responses of rat and mouse primary microglia to pro- and anti-inflammatory stimuli: molecular profiles, K(+) channels and migration.
复制标题
大鼠和小鼠原发性小胶质细胞对促炎和抗炎刺激的反应：分子特征，K（+）通道和迁移。
DOI：
10.1186/s12974-017-0941-3
复制
发表时间：
2017-08-22
影响因子：
9.3
通讯作者：
Schlichter LC
中科院分区：
文献类型：
--
作者：
Lam D;Lively S;Schlichter LC
急性中枢神经系统损伤通常使用大鼠和小鼠模型进行研究，但分子分析越来越多地发现物种差异可能会影响将研究结果转化为人类的能力。小胶质细胞可以经历复杂的分子和功能变化，通常通过体外对离散激活刺激的反应来研究。有相当多的证据表明，促炎(M1)激活可以加剧组织损伤，而抗炎(M2)状态有助于消炎和促进组织修复。然而，在评估控制炎症的潜在治疗靶点时，确定大鼠和小鼠的小胶质细胞是否有相同的反应是至关重要的。培养SD大鼠和C57BL/6小鼠的原代小胶质细胞，然后用干扰素-γ+肿瘤坏死因子-α(I+T；M1激活)、白介素4(M2a，交替激活)或IL-10(M2c，获得性失活)刺激。为了描述它们的激活反应，纳米串被用来监测许多促炎和抗炎介质、小胶质细胞标记物、免疫调节剂和其他分子的信使RNA(MRNA)的表达。Western分析用于检测选定的蛋白质。检测了控制炎症的两个潜在靶点-内向和外向整流性K+通道(Kir2.1，Kv1.3)，并应用特定的通道阻滞剂来确定它们在不同激活状态下对小胶质细胞迁移的贡献。I+T治疗后，促炎分子增加，但不同物种之间存在一些定性和定量的差异(例如，iNOS和一氧化氮，COX-2)。几个通常与M2a状态相关的分子在物种之间不同，或者它们是在额外的激活状态(例如CD206、ARG1)中诱导的。几种小胶质细胞标志物(IBA1、CD11b、CD68)的静息水平和/或反应因激活状态、物种或两者兼而有之而不同。在这两个物种中都检测到了几个Kir2和KV1家族成员的转录。然而，电流幅度(主要是Kir2.1和Kv1.3)取决于激活状态和物种。处理引起的形态和迁徙能力的变化在物种之间是相似的(迁徙减少I+T，增加IL-4或IL-10)。在这两个物种中，Kir2.1阻止迁移减少，Kv1.3阻止迁移增加，无论激活状态如何；因此，这些通道可能影响小胶质细胞向损伤部位的迁移。建议在概括小胶质细胞对物种间激活刺激的分子和功能反应时要谨慎。本文的在线版本(doi：10.1186/s12974-0170941-3)包含补充材料，授权用户可以使用。
Acute CNS damage is commonly studied using rat and mouse models, but increasingly, molecular analysis is finding species differences that might affect the ability to translate findings to humans. Microglia can undergo complex molecular and functional changes, often studied by in vitro responses to discrete activating stimuli. There is considerable evidence that pro-inflammatory (M1) activation can exacerbate tissue damage, while anti-inflammatory (M2) states help resolve inflammation and promote tissue repair. However, in assessing potential therapeutic targets for controlling inflammation, it is crucial to determine whether rat and mouse microglia respond the same. Primary microglia from Sprague-Dawley rats and C57BL/6 mice were cultured, then stimulated with interferon-γ + tumor necrosis factor-α (I + T; M1 activation), interleukin (IL)-4 (M2a, alternative activation), or IL-10 (M2c, acquired deactivation). To profile their activation responses, NanoString was used to monitor messenger RNA (mRNA) expression of numerous pro- and anti-inflammatory mediators, microglial markers, immunomodulators, and other molecules. Western analysis was used to measure selected proteins. Two potential targets for controlling inflammation—inward- and outward-rectifier K+ channels (Kir2.1, Kv1.3)—were examined (mRNA, currents) and specific channel blockers were applied to determine their contributions to microglial migration in the different activation states. Pro-inflammatory molecules increased after I + T treatment but there were several qualitative and quantitative differences between the species (e.g., iNOS and nitric oxide, COX-2). Several molecules commonly associated with an M2a state differed between species or they were induced in additional activation states (e.g., CD206, ARG1). Resting levels and/or responses of several microglial markers (Iba1, CD11b, CD68) differed with the activation state, species, or both. Transcripts for several Kir2 and Kv1 family members were detected in both species. However, the current amplitudes (mainly Kir2.1 and Kv1.3) depended on activation state and species. Treatment-induced changes in morphology and migratory capacity were similar between the species (migration reduced by I + T, increased by IL-4 or IL-10). In both species, Kir2.1 block reduced migration and Kv1.3 block increased it, regardless of activation state; thus, these channels might affect microglial migration to damage sites. Caution is recommended in generalizing molecular and functional responses of microglia to activating stimuli between species. The online version of this article (doi:10.1186/s12974-017-0941-3) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
9.3
作者：
Cherry JD;Olschowka JA;O'Banion MK
通讯作者：
O'Banion MK
影响因子：
9.3
作者：
Das A;Kim SH;Arifuzzaman S;Yoon T;Chai JC;Lee YS;Park KS;Jung KH;Chai YG
通讯作者：
Chai YG
影响因子：
3.8
作者：
Bowlby, MR;Fadool, DA;Levitan, IB
通讯作者：
Levitan, IB
影响因子：
6.2
作者：
Arnoux, Isabelle;Hoshiko, Maki;Audinat, Etienne
通讯作者：
Audinat, Etienne
影响因子：
6.3
作者：
Chen, Yi-Je;Nguyen, Hai M.;Wulff, Heike
通讯作者：
Wulff, Heike