数学分析法评估大蒜素对HeLa细胞氧化还原水平的影响

Effect of Allicin on the Redox Level of HeLa Cells as Assessed by Various Mathematical Analysis Methods

为探究大蒜素调控典型肿瘤细胞中的氧化还原水平,建立了佛波酯(phorbol-12-myristate-13-acetate,PMA)诱导人宫颈癌细胞HeLa中活性氧(reactiveoxygenspecies,ROS)升高的氧化应激模型,通过借助中心响应面模型,探究发现当HeLa细胞在10 mg/L的大蒜素培养液中孵育24 h时,其胞内ROS水平不会发生显著的上调。考虑到机体中的氧化还原水平处于复杂的动态变化中,因此分别对受到大蒜素调控的不同组HeLa细胞中的超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)、过氧化物酶(peroxidase,POD)、谷胱甘肽巯基转移酶(glutathione S-transferase,GST)等氧化还原生物标志物的活力进行测定,并建立基于层次分析法的数学分析模型,对HeLa细胞的抗氧化能力进行综合评估。结果认为,细胞中ROS在大蒜素调控的过程中,SOD、GSH-Px含量或许起到了主要的作用,并且10 mg/L大蒜素提前干预24 h后的HeLa细胞在抗氧化评估中获得了最高分,这与响应面模型筛选结果保持一致。最后,通过测定各组别中的丙二醛及8-氧鸟嘌呤脱氧核苷含量,得出大蒜素的抗氧化调控与保护或修复细胞中的脂质及DNA的过氧化损伤有关,并与综合评估结果呈现一定的相关性。采用多种数学分析法研究大蒜素对HeLa细胞氧化还原水平的调控作用,克服了生物体系中复杂体系的研究困难,经过推测与验证多种途径进行分析,为大蒜素保护细胞免受PMA氧化损伤的相关机理研究提供了参考。

In order to study redox regulation by allicin in typical tumor cells, an oxidative stress model was established using HeLa cells with increased reactive oxygen species(ROS) induced by phorbol-12-myristate-13-acetate(PMA). Using response surface methodology based on central composite design, it was found that the intracellular ROS level in HeLa cells was not be significantly up-regulated after incubation with 10 mg/L allicin for 24 h. Considering that the redox level in the body is in complex dynamic changes, the activities of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px),catalase(CAT), peroxidase(POD) and glutathione S-transferase(GST) as redox biomarkers in HeLa cells treated with different concentrations of allicin were determined, and a comprehensive mathematical model based on the analytic hierarchy process(AHP) was established to evaluate the antioxidant capacity of HeLa cells. The results showed that SOD and GSHPx may play major roles in the regulation of ROS in HeLa cells by allicin, and HeLa cells scored highest in antioxidant capacity evaluation after incubation with 10 mg/L allicin for 24 h, which was consistent with the results of the response surface model. Finally, by measuring the contents of malondialdehyde(MDA) and 8-oxo-2’-deoxyguanosine(8-oxo-dG) in each group, it was shown that the regulation of antioxidant capacity by allicin was related to its ability to protect and repair lipid peroxidation-induced DNA damage in cells, which was correlated with the comprehensive evaluation results. The application of a variety of mathematical analysis methods to study the regulation of allicin on the redox level of HeLa cells can overcome the difficulties encountered in research on complex biological systems. The results of this study provide a basis for further research on the mechanism of action of allicin in protecting cells from PMA-induced oxidative damage.