Sambucus ebulus L. Fruit Aqueous Infusion Modulates GCL and GPx4 Gene Expression

O. TASINOV, Y. KISELOVA-KANEVA and D. IVANOVA
Medical University “Prof. Dr. P. Stoyanov”, Department of Biochemistry, Molecular Medicine and Nutrigenomics, BG – 9002 Varna, Bulgaria

Abstract

Tasinov, O., Y. Kiselova-Kaneva and D. Ivanova, 2013.Sambucus ebulus L. fruit aqueous infusion modulates GCL and GPx4 gene expression. Bulg. J. Agric. Sci., Supplement 2, 19: 143–146

 

Glutamate-cysteine ligase and glutathione peroxidase 4 are enzymes involved in cellular antioxidant defense. Biologically active compounds, such as polyphenols, may modulate their gene expression. Sambucus ebulus L. is a plant, which fruit infusions exhibits high antioxidant activity, due to its high polyphenol content. We analyzed the effect of Sambucus ebulus fruit aqueous infusion on glutamate-cysteine ligase catalytic subunit and glutathione peroxidase 4 gene expression levels in the presence and in the absence of oxidizing agent tert-butyl hydroperoxide in a 3T3-L1 preadipocyte cell culture model. Gene expression was analyzed by Real-Time qPCR, and relative gene expression levels were calculated using 2–ΔΔCt method. Viability of treated preadipocytes was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid reduction assay. In the range of 1.25% to 12.5% content of the infusion in the culture medium cell viability was not affected significantly. Glutamate-cysteine ligase mRNA levels were increased significantly (p < 0.05) after treatment. Pretreatment decreased tert-butyl hydroperoxide-induced gene expression of glutamate-cysteine ligase (p < 0.05) and of glutathione peroxidase 4 (p < 0.05). These data suggest that Sambucus fruit infusion exhibits strong antioxidant activity by modulating glutamate-cysteine ligase and glutathione peroxidase 4 gene expression. The study provides first scientific data about the antioxidant activity of Sambucus ebulus fruit infusion in a cell culture model.

Key words: antioxidant, glutamate-cysteine ligase, glutathione peroxidase 4, Sambucus ebulus, 3T3-L1cells
Abbreviations: GCLc – glutamate-cysteine ligase catalytic subunit; GPx4 – glutathione peroxidase 4; GSH – gluthatione; SE – Sambucus ebulus; FAI – fruit aqueous infusion; tButOOH – tert-butyl hydroperoxide; Real-Time qPCR – Real Time quantitative polymerase chain reaction; ROS – reactive oxygen species; cAMP – cyclic adenosine monophosphate; PKA – protein kinase A; ARE/EpRE – аntioxidant response element/electrophile response element; CREB – cAMP response element binding protein

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