Barbisan Fernanda, Azzolin Verônica Farina, Ribeiro Euler Esteves, Duarte Marta Maria Medeiros Frescura, da Cruz Ivana Beatrice Mânica
1 Postgraduate Program of Pharmacology, Federal University of Santa Maria , Santa Maria, Brazil .
2 Open University of the Third Age, State University of Amazonas , Manaus, Brazil .
Rejuvenation Res. 2017 Aug;20(4):334-345. doi: 10.1089/rej.2016.1892. Epub 2017 May 10.
As superoxide is a key molecule of inflammatory activation, superoxide-hydrogen peroxide (S-HP) imbalance genetically caused could alter immunosenescence patterns. To test this hypothesis, we collected and cultured peripheral blood mononuclear cells (PBMCs) carrier's different genotypes of a genetic polymorphism located in the superoxide dismutase manganese-dependent gene (Val16Ala-SOD2). We used an in vitro genetic model based on previous studies, which suggested an association between homozygous genotypes (AA and VV) and alterations in oxidative-inflammatory mediators. PBMCs collected from young healthy volunteers were cultured in the presence of phytohemagglutinin, as well as the following cell culture passages obtained from the 72-hour initial culture. Each follow passage started with the same cell concentration (1 × 10 cells). The general immunosenescence pattern was observed independent of SOD2 genotypes: cellular proliferation until the 15th passage, when cellular arrestment occurred in the G0/G1 phase. From the 10th passage, a higher proliferative state was observed, indicating inflammatory hyperactivation, with an increase in the levels of inflammatory cytokines (IL-1, IL-6, and TNFα), nitric oxide, superoxide, lipoperoxidation, protein carbonylation, reactive oxygen species, and DNA damage. The S-HP imbalance affected the intensity of some immunosenescence parameters. AA cells, which present basal high HP levels, were associated with higher DNA damage and lipoperoxidation levels, whereas VV, which present basal high S levels, was associated with higher proinflammatory cytokine levels. In summary, the results suggested that a basal S-HP imbalance could affect the intensity of some immunosenescence markers, and this influence could explain the potential association between an imbalance of genotypes (AA and VV) and the risk of developing some chronic diseases.
由于超氧化物是炎症激活的关键分子，基因导致的超氧化物-过氧化氢（S-HP）失衡可能会改变免疫衰老模式。为了验证这一假设，我们收集并培养了外周血单个核细胞（PBMC），其携带者为位于锰依赖性超氧化物歧化酶基因（Val16Ala-SOD2）的不同基因型的遗传多态性。我们基于先前的研究使用了一种体外遗传模型，该研究表明纯合基因型（AA和VV）与氧化-炎症介质的改变之间存在关联。从年轻健康志愿者收集的PBMC在植物血凝素存在的情况下进行培养，以及从72小时初始培养中获得的以下细胞培养传代。每次后续传代都从相同的细胞浓度（1×10个细胞）开始。观察到一般的免疫衰老模式与SOD2基因型无关：细胞增殖直至第15代，此时细胞在G0/G1期停滞。从第10代开始，观察到更高的增殖状态，表明炎症过度激活，炎症细胞因子（IL-1、IL-6和TNFα）、一氧化氮、超氧化物、脂质过氧化、蛋白质羰基化、活性氧和DNA损伤水平增加。S-HP失衡影响了一些免疫衰老参数的强度。具有基础高HP水平的AA细胞与更高的DNA损伤和脂质过氧化水平相关，而具有基础高S水平的VV细胞与更高的促炎细胞因子水平相关。总之，结果表明基础S-HP失衡可能会影响一些免疫衰老标志物的强度，这种影响可以解释基因型（AA和VV）失衡与某些慢性疾病发生风险之间的潜在关联。
