The Investigation of DNA Damage Induced by Adrenaline in Human Lymphocytes in Vitro/Ispitivanja Oštećenja DNK Izazvanih Adrenalinom U Limfocitima Čoveka in Vitro

Abstract Adrenaline is a neurotransmitter and hormone that plays an important role in physiological regulatory mechanisms. The objective of this study was to assess primary DNA damage in isolated human lymphocytes exposed to adrenaline using the in vitro comet assay. Dose-response of human lymphocytes was determined at concentration range of adrenaline from 0.01 μM to 300 μM for various treatment times (1h, 2h, 4h and 24h). The obtained results showed that adrenaline induced DNA damage at concentration range from 5 μM to 300 μM after 1h, 2h and 4h of treatment. The slightest DNA damage was observed after 24 h of adrenaline treatment - only the highest concentrations of adrenaline (150 μM and 300 μM) caused increased level of DNA damage. In order to evaluate the potential contribution of reactive oxygen species (ROS) in adrenaline-induced DNA damage we used antioxidants catalase (100 IU/mL and 500 IU/mL) and quercetin (100 μM and 500 μM). Co-treatment of lymphocytes with adrenaline (300 μM) and antioxidants for 1 h, significantly reduced the quantity of DNA in the comet tails. Therefore, it can be concluded that adrenaline exhibits genotoxic effects mainly through induction of reactive oxygen species and that some of the DNA damage is repaired during the first four hours following the treatment with adrenaline.

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Niosome-carvedilol protects DNA damage of supraphysiologic concentrations of insulin using comet assay: An in vitro study

Human & Experimental Toxicology ◽

10.1177/09603271211036124 ◽

2021 ◽

pp. 096032712110361

Author(s):

Marzieh Farahani-Zangaraki ◽

Azade Taheri ◽

Mahmoud Etebari

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Dna Damage ◽

Comet Assay ◽

Protective Effect ◽

Reactive Oxygen ◽

Diabetic Patients ◽

Oxygen Species ◽

Type 2 Diabetic Patients

Introduction: Hyperinsulinemia occurs in type 2 diabetic patients with insulin resistance. This increase in insulin levels in the blood increases reactive oxygen species production and oxidative stress, resulting in DNA damage. Carvedilol (CRV) is a non-selective beta-blocker, and research has shown that this compound and its metabolites have anti-oxidative properties. Carvedilol can, directly and indirectly, reduce reactive oxygen species (ROS) and has a protective effect on DNA damage from oxidative stress. Given the insolubility of CRV in water, finding new methods to increase its solubility can be an essential step in research. This study aimed to determine whether carvedilol could have a protective effect on insulin-induced genomic damage. Methods: We treated cells with insulin alone, amorphous-CRV alone, and amorphous-CRV and niosomal-CRV with insulin and DNA damage were investigated using the comet method to achieve this goal. Results: Our results showed that insulin in the studied concentration has a significant genotoxic effect and non-cytotoxic at higher concentrations. CRV, both in amorphous and niosome form, reduced insulin-induced DNA damage by reducing ROS production. The comet assay results demonstrate that treating HUVEC cells in pretreatment condition with amorphous-CRV and niosome-CRV significantly reduces DNA damage of insulin.

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