Cytotoxic effects of two extracts from garlic (Allium sativum L.) cultivars on the human squamous carcinoma cell line SCC-15

AbstractGarlic (Allium sativum L., Alliaceae) has acquired a reputation as a therapeutic agent and herbal remedy to prevent and treat several pathologies. The aim of the present study was to determine the effect of two Allium sativum L. cultivars, Harnaś and Morado, on reactive oxygen species (ROS) production, viability and apoptotic process in human squamous carcinoma cell line SCC-15. The experiments were conducted on SCC-15 cell line exposed to increasing concentrations of garlic extracts of 0.062, 0.125, 0.250, 0.500 and 1.000mg/mL. After the experiments, ROS formation, caspase-3 activity and neutral red uptake were measured in the cells, and in a collected medium lactate dehydrogenase (LDH) release was measured. The Spanish cultivar Morado has demonstrated higher potential to stimulate ROS production in SCC-15 cells after a short time period (6h) than the Polish cultivar Harnaś. However, the Polish cultivar Harnaś manifested more prolonged potential to stimulate ROS production in SCC-15 cells. Both studied garlic extracts induced cytotoxicity on SCC-15 cell line which was probably ROS-dependent. We also determined that in SCC-15 cells high concentrations of studied extracts did not cause activation of caspase-3 which suggested caspase-independent or necrotic cell death

Review of the Relationship between Reactive Oxygen Species (ROS) and Elastin-Derived Peptides (EDPs)

Reactive oxygen species (ROS) are central elements of a number of physiological processes such as differentiation and intracellular signaling, as well as pathological processes, e.g., inflammation or apoptosis. ROS are involved in the growth and proliferation of stem cells, cell communication, cell aging, all types of inflammation, cancer development and proliferation, or type 2 diabetes. Elastin-derived peptides (EDPs) are detected in all these conditions and, according to the current state of knowledge, the role of the extracellular matrix (ECM) protein is crucial. It is believed that EDPs are a result of the aforementioned pathological conditions and are generated during degradation of ECM. However, as shown in the literature, the production of EDPs can be induced not only by inter alia chemical, enzymatic, and physical factors but also directly by ROS. No comprehensive study of the impact of ROS on EDPs and EDPs on ROS production has been conducted to date; therefore, the aim of this paper is to summarize the current state of knowledge of the relationship between ROS and ECM with special involvement of EDPs in the processes mentioned above. Depending on the type of cells, tissue, or organism, the relationships between ROS and ECM/EDPs may differ completely

Methods of Isolation of Active Substances from Garlic (Allium sativum L.) and Its Impact on the Composition and Biological Properties of Garlic Extracts

Garlic (Allium sativum L.) is widely used in the human diet and in scientific research due to its biological properties. Various factors, e.g., temperature, pressure, extraction method, type of solvent, size, and territorial origin of garlic, affect the amount and type of bioactive compounds obtained from garlic extracts. In turn, the content of bioactive compounds correlates with the biological activity of the extracts. Therefore, the aim of this review was to summarize the current state of knowledge of the methods and effectiveness of isolation of active substances from garlic and their impact on the garlic extract composition and, consequently, biological properties. According to the literature, extracts obtained using water as a solvent are mainly responsible for antimicrobial properties, which is related to, inter alia, the high content of allicin. The use of alcohols, such as methanol or ethanol, is associated with the outstanding antioxidant power of extracts resulting from the presence of phenolic compounds. In turn, due to the presence of diallyl disulfide and disulfide trisulfide, garlic oil has anticancer potential. Acetone is the most effective organic solvent; however, it is not suitable for immediate consumption

Possibility to Biotransform Anthracyclines by Peroxidases Produced by Bjerkandera adusta CCBAS 930 with Reduction of Geno- and Cytotoxicity and Pro-Oxidative Activity

The aim of this study was to evaluate the bioremoval mechanism of anthracycline antibiotics by the white-rot fungus B. adusta CCBAS 930. The activity of oxidoreductases and levels of phenolic compounds and free radicals were determined during the biotransformation of anthraquinone antibiotics: daunomycin (DNR) and doxorubicin (DOX) by B. adusta strain CCBAS 930. Moreover, phytotoxicity (Lepidium sativum L.), ecotoxicity (Vibrio fischeri), genotoxicity and cytotoxicity of anthraquinone dyes were evaluated before and after biological treatment. More than 80% and 90% of DNR and DOX were removed by biodegradation (decolorization). Initial solutions of DNR and DOX were characterized by eco-, phyto-, geno- and cytotoxicity. Despite efficient decolorization, secondary metabolites, toxic to bacteria, formed during biotransformation of anthracycline antibiotics in B. adusta CCBAS 930 cultures. DNR and DOX metabolites did not increase reactive oxygen species (ROS) production in human fibroblasts and resazurin reduction. DNR metabolites did not change caspase-3 activity

Possibility to Biotransform Anthracyclines by Peroxidases Produced by <i>Bjerkandera adusta</i> CCBAS 930 with Reduction of Geno- and Cytotoxicity and Pro-Oxidative Activity

The aim of this study was to evaluate the bioremoval mechanism of anthracycline antibiotics by the white-rot fungus B. adusta CCBAS 930. The activity of oxidoreductases and levels of phenolic compounds and free radicals were determined during the biotransformation of anthraquinone antibiotics: daunomycin (DNR) and doxorubicin (DOX) by B. adusta strain CCBAS 930. Moreover, phytotoxicity (Lepidium sativum L.), ecotoxicity (Vibrio fischeri), genotoxicity and cytotoxicity of anthraquinone dyes were evaluated before and after biological treatment. More than 80% and 90% of DNR and DOX were removed by biodegradation (decolorization). Initial solutions of DNR and DOX were characterized by eco-, phyto-, geno- and cytotoxicity. Despite efficient decolorization, secondary metabolites, toxic to bacteria, formed during biotransformation of anthracycline antibiotics in B. adusta CCBAS 930 cultures. DNR and DOX metabolites did not increase reactive oxygen species (ROS) production in human fibroblasts and resazurin reduction. DNR metabolites did not change caspase-3 activity

Elastin-derived peptide VGVAPG decreases differentiation of mouse embryo fibroblast (3T3-L1) cells into adipocytes

Elastin is a highly elastic protein present in connective tissue. As a result of protease activity, elastin hydrolysis occurs, and during this process, elastin-derived peptides (EDPs) are released. One of the constitutively repeating elastin and EDP building sequences is the hexapeptide VGVAPG. Therefore, the aim of our research was to define the effect of VGVAPG peptide on adipogenesis in a mouse 3T3-L1 cell line. 3T3-L1 cells were differentiated according to a previously described protocol and exposed to increasing concentrations of VGVAPG or VVGPGA peptide. The obtained results showed that VGVAPG peptide does not stimulate reactive oxygen species (ROS) production, caspase-1 activation, and 3T3-L1 cell proliferation. In the second part of the experiments, it was proved that VGVAPG peptide decreased lipid accumulation as measured by oil red O staining, which was confirmed by the profile of increased expression markers of undifferentiated preadipocytes. In our experiments, 10 nM VGVAPG added for differentiating to adipocytes increased the expression of Pref-1, serpin E1, and adiponectin as compared to rosiglitazone (PPARγ agonist)-treated group and simultaneously decreased the expression of VEGF and resistin as compared to the rosiglitazone-treated group. The obtained results show that VGVAPG peptide sustains 3T3 cells in undifferentiated state. Abbreviations: DMSO: dimethyl sulphoxide; EBP: elastin-binding protein; EDPs: elastin-derived peptides; FBS: foetal bovine serum; Glb1: gene for beta-galactosidase; LDL: low-density-lipoprotein; PAI-1 (Serpin E1): plasminogen activator inhibitor-1; PBS: phosphate-buffered saline; PPARγ: peroxisome proliferator-activated receptor gamma; Pref-1: preadipocyte factor 1; ROS: reactive oxygen species; VEGF-A: vascular endothelial growth factor-A; VGVAPG: Val-Gly-Val-Ala-Pro-Gly; β-Gal: beta-galactosidase; ORO: oil red O; IBMX: 3-isobutyl-1-methylxanthine; H2DCFDA: 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate; DMEM: Dulbecco’s Modified Eagle’s Medium; VVGPGA: Val-Val-Gly-Pro-Gly-Ala