Barbisan Fernanda, Azzolin Verônica Farina, Monteiro Gustavo Cardenas, Teixeira Cibele F, Mastella Moisés Henrique, Bueno Vitor, Duarte Marta Maria Medeiros Frescura, Wagner Glauber, do Prado-Lima Pedro Antônio Schmidt, Ribeiro Euler Esteves, da Cruz Ivana B M
Postgraduate Program of Gerontology, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Postgraduate Program of Pharmacology, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Gene. 2018 May 20;655:48-55. doi: 10.1016/j.gene.2018.02.046. Epub 2018 Feb 18.
Lithium (Li), a mood stabilizer used to treat bipolar disorder (BP) symptoms has important anti-inflammatory effects by downregulation of glycogen synthase kinase-3 beta (GSK-3β). However, sometime Li effect is not efficient in some patients suggesting genetic interference. Previous investigations described association between a genetic superoxide‑hydrogen (S-HP) imbalance caused by a superoxide dismutase manganese dependent gene polymorphism (Val16Ala-SOD2 SNP, rs4880) and differential anti-inflammatory response of some drugs and bioactive molecules. Therefore, we postulated here that S-HP imbalance could present some effect on GSK-3β modulation by Li.
to test this hypothesis, a genetic and a pharmacological S-HP imbalance protocols were performed. In the two protocols, immune cells were activated by phythohemaglutin (PHA). The first one, used peripheral blood mononuclear cells (PBMCs) cultures carrying different Val16Ala-SOD2 genotypes, and the second used a commercial macrophage cell line RAW 264.7. Macrophages were exposed to paraquat to induce high S levels (VV-like cells) or porphyrin, that is a SOD2-like molecule that increase dismutation of S into HP (AA-like cells). In both protocols the Li effects on GSK-3β gene and protein modulation as evaluated in 24 h cultures. The inflammatory activation was also analyzed by cellular proliferation in 72 h cell cultures.
as expected PHA exposure triggered a strong upregulation of GSK-3β gene expression (p ≤ 0.001), and Li exposure showed GSK-3β gene downregulation from 0.7 mEq/L concentrations. However, Li modulatory effects on GSk-3β gene and protein expression was directly influenced by basal S-HP balance. Presence of high S-basal levels (VV genotype and VV-like cells) induced attenuated Li anti-inflammatory effects in comparison with balanced and AA and AA-like cells (p < 0.001). Despite methodological limitations related to in vitro assays, the whole of results suggested that Li anti-inflammatory effects is influenced by S-HP basal state and is plausible that its influence could contributes to resistance of some patients to Li treatment or to increase of intensity of some side effects Li-associated.
锂（Li）是一种用于治疗双相情感障碍（BP）症状的情绪稳定剂，通过下调糖原合酶激酶-3β（GSK-3β）具有重要的抗炎作用。然而，有时锂在某些患者中效果不佳，提示存在基因干扰。先前的研究描述了由超氧化物歧化酶锰依赖性基因多态性（Val16Ala-SOD2 SNP，rs4880）引起的遗传超氧化物-过氧化氢（S-HP）失衡与某些药物和生物活性分子的不同抗炎反应之间的关联。因此，我们在此推测S-HP失衡可能对锂对GSK-3β的调节产生某些影响。
为了验证这一假设，进行了遗传和药理学S-HP失衡实验方案。在这两个实验方案中，免疫细胞通过植物血凝素（PHA）激活。第一个方案使用携带不同Val16Ala-SOD2基因型的外周血单核细胞（PBMC）培养物，第二个方案使用商业巨噬细胞系RAW 264.7。巨噬细胞暴露于百草枯以诱导高S水平（VV样细胞）或卟啉，卟啉是一种类似SOD2的分子，可增加S向HP的歧化（AA样细胞）。在两个实验方案中，均在24小时培养物中评估锂对GSK-3β基因和蛋白质调节的影响。还通过72小时细胞培养物中的细胞增殖分析炎症激活情况。
正如预期的那样，PHA暴露引发了GSK-3β基因表达的强烈上调（p≤0.001），锂暴露在0.7mEq/L浓度时显示出GSK-3β基因下调。然而，锂对GSK-3β基因和蛋白质表达的调节作用直接受到基础S-HP平衡的影响。与平衡状态以及AA和AA样细胞相比，高S基础水平（VV基因型和VV样细胞）的存在导致锂的抗炎作用减弱（p<0.001）。尽管与体外试验相关存在方法学局限性，但整体结果表明锂的抗炎作用受S-HP基础状态影响，并且其影响可能导致一些患者对锂治疗产生耐药性或增加与锂相关的某些副作用的强度，这似乎是合理的。
