Programa de Pós-Graduação em Bioquímica, Universidade Federal do Pampa, Uruguaiana, Brazil.
Curso de Farmácia, Universidade Federal do Pampa, Uruguaiana, Brazil.
BMC Pharmacol Toxicol. 2019 Dec 19;20(Suppl 1):79. doi: 10.1186/s40360-019-0359-x.
The γ-hexalactone is a flavoring agent for alcoholic beverages, teas, breads, dairy products, coffees, buttery products among others. It presents low molecular weight and exhibits sweet fruity aroma with nuances of nuts. As far as we know, both literature and government regulations have gaps regarding the safe use of the γ-hexalactone. In this context, the main objective of this work was to evaluate the effects of γ-hexalactone through in silico and in vitro approaches.
The in silico analysis was performed through four free online platforms (admetSAR, Osiris Property Explorer, pkCSM platform and PreADMET) and consisted of comparative structural analysis with substances present in databases. The computational prediction was performed in the sense of complement and guide the in vitro tests. Regarding in vitro investigations, screening of cytotoxicity (assessed by cell proliferation and viability parameters) in lymphocytes exposed to γ-hexalactone for 72 h were carried out previously to determine non-cytotoxic concentrations. Following this screening, concentrations of 5.15, 0.515, and 0.0515 μM were selected for the study of the respective potentials: genotoxic (assessed by DNA comet assay), chromosomal mutation (analysis of micronucleus frequency) and immunomodulatory (cytokine quantification using ELISA immunoassay). The results of in vitro assays were compared by one-way analysis of variance (ANOVA), followed by Bonferroni's post hoc test, conducted by statistic software.
The platform PreADMET pointed out that γ-hexalactone is potentially mutagenic and carcinogenic. The comet assay data corroborate with these results demonstrating that γ-hexalactone at 5.15 μM caused lymphocytes DNA damage. In relation to cytokine secretion, the results indicate that lymphocytes were activated by γ-hexalactone at non-cytotoxic concentrations, involving an increase in the IL-1 levels in all tested concentrations, ranging from approximately 56 to 93%. The γ-hexalactone only at 5.15 μM induced increase in the levels of IL-6 (~ 60%), TNF-α (~ 68%) and IFN-γ (~ 29%), but decreased IL-10 (~ 46%) in comparison with the negative control (p < 0.05). No change was observed in total lymphocytes or in cell viability at the concentrations tested.
In summary, the γ-hexalactone demonstrated immunomodulatory and genotoxic effects at non-cytotoxic concentrations in healthy lymphocytes.
γ-己内酯是一种用于酒精饮料、茶、面包、乳制品、咖啡、黄油产品等的调味剂。它的分子量低，具有甜果味香气，并带有坚果的细微差别。据我们所知，文献和政府法规在 γ-己内酯的安全使用方面都存在空白。在这种情况下，这项工作的主要目的是通过计算和体外方法评估 γ-己内酯的影响。
通过四个免费在线平台（admetSAR、Osiris Property Explorer、pkCSM 平台和 PreADMET）进行计算分析，包括与数据库中存在的物质进行比较结构分析。计算预测是为了补充和指导体外测试而进行的。关于体外研究，在确定非细胞毒性浓度之前，用 γ-己内酯处理淋巴细胞 72 小时，进行细胞增殖和活力参数评估的细胞毒性筛选。在此筛选之后，选择 5.15、0.515 和 0.0515 μM 的浓度用于研究各自的潜力：遗传毒性（通过 DNA 彗星试验评估）、染色体突变（微核频率分析）和免疫调节（ELISA 免疫分析测定细胞因子定量）。通过统计软件进行单向方差分析（ANOVA），然后进行 Bonferroni 事后检验，比较体外试验的结果。
PreADMET 平台指出，γ-己内酯具有潜在的致突变性和致癌性。彗星试验数据与这些结果相符，表明 5.15 μM 的 γ-己内酯导致淋巴细胞 DNA 损伤。关于细胞因子分泌，结果表明，γ-己内酯在非细胞毒性浓度下激活淋巴细胞，所有测试浓度的 IL-1 水平均增加，约为 56%至 93%。γ-己内酯仅在 5.15 μM 时诱导 IL-6（60%）、TNF-α（68%）和 IFN-γ（29%）水平升高，但与阴性对照相比，IL-10 水平降低（46%）（p<0.05）。在测试浓度下，总淋巴细胞或细胞活力没有变化。
总之，γ-己内酯在健康淋巴细胞的非细胞毒性浓度下表现出免疫调节和遗传毒性作用。
