Abstract：
Further understanding the mechanism for microglia activation is necessary for developing novel anti-inflammatory strategies. Our previous study found that the activation of sigma-1 receptor can effectively inhibit the neuroinflammation, independent of the canonical mechanisms, such as NF-κB, JNK and ERK inflammatory pathways. Thus, it is reasonable that an un-identified, non-canonical pathway contributes to the activation of microglia. In the present study, we found that a sigma-1 receptor agonist of 2-morpholin-4-ylethyl 1-phenylcyclohexane-1-carboxylate (PRE-084) suppressed lipopolysaccharide (LPS) elevated nitric oxide (NO) content in BV-2 microglia culture supernatant and LPS-raised mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS) in BV-2 microglia. Moreover, PRE-084 alleviated LPS-increased Ser 9 de-phosphorylation of glycogen synthase kinase-3 beta (GSK-3β), LPS-elevated catalytic activity of calcineurin, and LPS-raised percent and frequency of Ca2+ oscillatory BV-2 cells. We further found that the inhibitory effect of PRE-084 was reversed by a calcineurin activator of chlorogenic acid and a GSK-3β activator of pyrvinium. Moreover, an IP3 receptor inhibitor of 2-aminoethoxydiphenyl borate mimicked the anti-inflammatory activity of PRE-084. Thus, we identified a noncanonical pro-neuroinflammary pathway of Ca2+ oscillation/Calcineurin/GSK-3β and the inhibition of this pathway is necessary for the anti-inflammatory activity of sigma-1 receptor activation.
摘要：
进一步了解小胶质细胞激活的机制对于开发新的抗炎策略是必要的。我们之前的研究发现激活sigma-1受体能有效抑制神经炎症,独立于规范机制，如NF-κB，JNK和ERK炎症途径。因此，一个身份不明的人是合理的，非经典途径有助于小胶质细胞的活化。在本研究中,我们发现2-吗啉-4-乙基1-苯基环己烷-1-羧酸（PRE-084）的sigma-1受体激动剂抑制脂多糖（LPS）升高BV-2小胶质细胞培养上清液中一氧化氮（NO）的含量和LPS升高的肿瘤坏死因子-α（TNF-α）的mRNA水平，白细胞介素-1β（IL-1β），BV-2小胶质细胞中的诱导型一氧化氮合酶（iNOS）。此外，PRE-084减轻了LPS增加的糖原合酶激酶-3β（GSK-3β）的Ser9去磷酸化，LPS提高钙调磷酸酶的催化活性，和LPS升高Ca2振荡BV-2细胞的百分比和频率。我们进一步发现，绿原酸的钙调磷酸酶激活剂和吡咯的GSK-3β激活剂可以逆转PRE-084的抑制作用。此外，2-氨基乙氧基二苯基硼酸盐的IP3受体抑制剂模拟PRE-084的抗炎活性。因此，我们确定了Ca2+振荡/钙调磷酸酶/GSK-3β的非经典促神经炎症途径,并且该途径的抑制对于sigma-1受体激活的抗炎活性是必需的.
