Objective: This study aimed to determine the capability of newly designed 3-methoxy derivatives of salicylaldehyde benzoylhydrazone to influence the oxidative stress processes and to test their in vitro cytotoxicity.

Methods: We have used chemiluminescent and spectrophotometric model systems containing different types of reactive oxygen species (OH^●, OCl^─ and O₂^─●). The hydrazones effect on the viability of Hep-G2, HEK-293 and SH-SY5Y cell lines was determined via MTT assay.

Results: The comparative analysis of the C₅₀ values of the chemiluminescent investigation demonstrated moderate activity against the hydroxyl radicals (C₅₀ > 50 μmol/L) and remarkable reactivity in the systems containing a superoxide radical and a hypochlorous anion (C₅₀ < 3.7 μmol/L). Further experiments in the spectrophotometric system of UV-induced OH^● generation and consequent 2′-deoxyribose oxidative damage excluded the possibility of quenching effect and proved the direct interaction of the studied compounds with that generated in the system reactive oxygen species (ROS). The encapsulation of the studied derivatives into chitosan-alginate particles led to the protection and stabilization of their antioxidant activity as revealed by a one-month study using the ABTS ^●+ method. The cytotoxic study revealed less pronounced effects against the non-malignant cell line (HEK-293) compared to Hep-G2 and SH-SY-5Y cells.

Discussion: The incorporation of a hydroxyl group in the hydrazide part of a parent molecule which relates to better antioxidant effect in most of the studied systems is associated with higher IC₅₀ values in all cytotoxicity experiments and relates to the cytoprotective effect against N-methyl-D-aspartate-induced excitotoxicity in SH-SY5Y human neuronal cells.

目的：本研究旨在确定新设计的水杨醛苯甲酰hydr的3-甲氧基衍生物影响氧化应激过程并测试其体外细胞毒性的能力。

方法：我们已经使用化学发光和含有不同类型的反应性氧物种的分光光度模型系统（OH ^●，OCL ^─和O ₂ ^─●）。通过MTT分析确定对Hep-G2，HEK-293和SH-SY5Y细胞系活力的影响。

结果：化学发光研究的C ₅₀值的对比分析表明，对羟基自由基的活性适中（C ₅₀  > 50μmol/ L），并且在含有超氧自由基和次氯阴离子的系统中（C ₅₀  <3.7μmol ）具有显着的反应活性/ L）。UV诱导的分光光度法中的进一步实验^●生成和随后的2'-脱氧核糖氧化损伤排除了淬灭作用的可能性，并证明了所研究化合物与系统活性氧（ROS）中所产生化合物的直接相互作用。一项为期一个月的使用ABTS ^●+方法的研究表明，将研究的衍生物封装到壳聚糖-海藻酸盐颗粒中可导致其抗氧化活性的保护和稳定。细胞毒性研究显示，与Hep-G2和SH-SY-5Y细胞相比，对非恶性细胞系（HEK-293）的作用不太明显。

讨论：在大多数研究的系统中，在母体分子的酰肼部分掺入羟基与更好的抗氧化作用有关，在所有细胞毒性实验中均与较高的IC ₅₀值相关，并与针对N-甲基- D-天冬氨酸诱导的SH-SY5Y人神经元细胞的兴奋性毒性。