Hansson Elisabeth, Werner Tony, Björklund Ulrika, Skiöldebrand Eva
Department of Clinical Neuroscience, Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg, SE 413 45, Gothenburg, Sweden.
Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
IBRO Rep. 2016 Jun 23;1:1-9. doi: 10.1016/j.ibror.2016.06.001. eCollection 2016 Dec.
This study aimed to test pharmaceutical compounds targeting astrocytes showing inflammatory dysregulation. The primary rat brain cultures were treated with different batches of serum with or without microglia added to make the cells inflammatory-reactive. Lipopolysaccharide (LPS) and tryptase were used as inflammatory inducers. Expression levels of Toll-like receptor 4 (TLR4), Na/K-ATPase, and matrix metalloprotease-13 (MMP-13), as well as actin filament organization, pro-inflammatory cytokines, and intracellular Ca release, were evaluated. LPS combined with tryptase upregulated TLR4 expression, whereas Na/K-ATPase expression was downregulated, ATP-evoked Ca transients were increased, actin filaments were reorganized and ring structures instead of stress fibers were observed. Other aims of the study were to prevent astrocytes from becoming inflammatory-reactive and to restore inflammatory dysregulated cellular changes. A combination of the μ-opioid antagonist (-)-naloxone in ultra-low concentrations, the non-addictive μ-opioid agonist (-)-linalool, and the anti-epileptic agent levetiracetam was examined. The results indicated that this drug cocktail prevented the LPS- and tryptase-induced inflammatory dysregulation. The drug cocktail could also restore the LPS- and tryptase-treated cells back to a normal physiological level in terms of the analyzed parameters.
本研究旨在测试针对表现出炎症调节异常的星形胶质细胞的药物化合物。用不同批次的血清处理原代大鼠脑培养物，添加或不添加小胶质细胞以使细胞产生炎症反应。脂多糖（LPS）和类胰蛋白酶用作炎症诱导剂。评估了Toll样受体4（TLR4）、钠钾ATP酶、基质金属蛋白酶-13（MMP-13）的表达水平，以及肌动蛋白丝的组织、促炎细胞因子和细胞内钙释放情况。LPS与类胰蛋白酶联合上调了TLR4的表达，而钠钾ATP酶的表达下调，ATP诱发的钙瞬变增加，肌动蛋白丝重新组织，观察到环状结构而非应力纤维。该研究的其他目的是防止星形胶质细胞产生炎症反应，并恢复炎症调节异常的细胞变化。研究了超低浓度的μ-阿片受体拮抗剂（-）-纳洛酮、非成瘾性μ-阿片受体激动剂（-）-芳樟醇和抗癫痫药物左乙拉西坦的组合。结果表明，这种药物组合可预防LPS和类胰蛋白酶诱导的炎症调节异常。就分析的参数而言，该药物组合还可使经LPS和类胰蛋白酶处理的细胞恢复到正常生理水平。
