Protutumorska i antioksidativna svojstva terpinolena u moždanih stanica štakora

Terpinolene (TPO) is a natural monoterpene present in essential oils of many aromatic plant species. Although various biological activities of TPO have been demonstrated, its neurotoxicity has never been explored. In this in vitro study we investigated TPO’s antiproliferative and/or cytotoxic properties using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) test, genotoxic damage potential using the single-cell gel electrophoresis (SCGE), and oxidative effects through total antioxidant capacity (TAC) and total oxidative stress (TOS) in cultured primary rat neurons and N2a neuroblastoma cells. Dose-dependent effects of TPO (at 10 mg L-1, 25 mg L-1, 50 mg L-1, 100 mg L-1, 200 mg L-1, and 400 mg L-1) were tested in both cell types. Significant (P<0.05) decrease in cell proliferation were observed in cultured primary rat neurons starting with the dose of 100 mg L-1 and in N2a neuroblastoma cells starting with 50 mg L-1. TPO was not genotoxic in either cell type. In addition, TPO treatment at 10 mg L-1, 25 mg L-1, and 50 mg L-1 increased TAC in primary rat neurons, but not in N2a cells. However, at concentrations above 50 mg L-1 it increased TOS in both cell types. Our findings clearly demonstrate that TPO is a potent antiproliferative agent for brain tumour cells and may have potential as an anticancer agent, which needs to be further studied.Terpinolen (TPO) prirodni je monoterpen prisutan u esencijalnim uljima mnogih aromatskih biljaka. Premda su otprije poznate razne biološke aktivnosti TPO-a, dosad nije ispitana njegova neurotoksičnost. Svrha je ovog istraživanja in vitro bila utvrditi antiproliferacijska i/ili citotoksična svojstva TPO-a pomoću testa 3-(4,5-dimetiltiazol-2-yl)-2,5 difeniltetrazolijeva bromida (MTT), njegov genotoksični potencijal pomoću komet-testa te oksidativno djelovanje kroz ukupni antioksidativni kapacitet i ukupni oksidativni stres u uzgojenim primarnim neuronima štakora i N2a stanicama neuroblastoma. U objema staničnim linijama ispitani su učinci TPO-a u skladu sa sljedećim dozama: 10 mg L-1, 25 mg L-1, 50 mg L-1, 100 mg L-1, 200 mg L-1 i 400 mg L-1. Značajni (p<0.05) pad stanične proliferacije u primarnim neuronima štakora zamijećen je pri dozama od 100 mg L-1 naviše, a u N2a stanicama neuroblastoma pri dozama od 50 mg L-1 naviše. Niti u jednoj staničnoj liniji TPO se nije pokazao genotoksičnim. Usto se primjenom TPO-a pri dozama od 10 mg L-1, 25 mg L-1 i 50 mg L-1 povećao ukupni antioksidativni kapacitet primarnih štakorskih neurona, ali je takvo djelovanje izostalo u N2a stanica. Međutim, pri koncentracijama višim od 50 mg L-1 TPO je povećao ukupni oksidativni stres u objema staničnim linijama. Naši rezultati nedvojbeno pokazuju da je TPO snažan antiproliferacijski agens u tumorskih stanica mozga, a njegovu potencijalnu ulogu kao protutumorskog lijeka trebalo bi dalje istraživati

The genoprotective activity of resveratrol on permethrin-induced genotoxic damage in cultured human lymphocytes

The aim of this work was to investigate the genetic effects of resveratrol (RSV) at concentrations of 10, 15, 25, 40, 75 and 100 µM and its activities on the genotoxicity induced by the permethrin (PM) (200 µM). After the application of PM and RSV, separately and together, cultured human lymphocytes were assessed by chromosome aberrations (CA) and sister chromatid exchange (SCE) tests. According to results, the frequencies of CA and SCE rates in the peripheral lymphocytes were significantly increased by PM compared with the controls. However, RSV had no genotoxic effect. Furthermore, the findings revealed that PM-induced increases in the mean frequencies of both genotoxic indices were diminished by RSV in a clear dose dependent manner, indicating its protective role towards the cells from PM exerted injury. In conclusion, these effects of RSV should be considered while evaluating the possible use of RSV as a therapeutic agent

An antidote for imazalil-induced genotoxicity in vitro: The lichen, Dermatocarpon intestiniforme (Körber) hasse

Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce serious toxic effects in vertebrates. In recent years, a number of studies have suggested that lichens might be easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being performed to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. In this context, the antigenotoxic effect of aqueous Dermatocarpon intestiniforme (Körber) Hasse. extract (DIE) was studied against the genotoxic damage induced by IMA on cultured human lymphocytes (n = 6) using chromosomal aberration (CA) and micronucleus (MN) as cytogenetic endpoints. Human peripheral lymphocytes were treated in vitro with varying concentrations of DIE (0, 25, 50 and 100 μg/ml), tested in combination with IMA (336 μg/ml). DIE alone were not genotoxic and when combined with IMA treatment, it reduced the frequency of CAs and the rate of MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of DIE. The results of the present study suggest that this plant extract per se does not have a genotoxic potential, but can alleviate the genotoxicity of IMA on cultured human lymphocytes. In conclusion our findings may have an important application for the protection of cultured human lymphocyte from the genetic damage and side effects induced by medical and agricultural chemicals hazardous for people

CYTOTOXICITY AND GENOTOXICITY OF IRON OXIDE NANOPARTICLES: AN IN VITRO BIOSAFETY STUDY

With the development of nanotechnology and the wide use of iron oxide nanoparticles, it has become necessary to assess the potential adverse biological effects of magnetite. This study investigated the cytotoxicity, genotoxicity and oxidative damage of different concentrations of magnetite (0 to 1000 mg/L) in human whole blood cultures. After supplementation of magnetite, the blood samples were incubated for 72 h. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) release assays. The total antioxidant capacity (TAC) and total oxidant status (TOS) were determined to evaluate the dose-dependent effects of magnetite on the oxidant/antioxidant balance and to evaluate the potential oxidative injury due to increased oxidative stress. Genotoxicity was estimated by by the sister chromatid exchange (SCE), micronuclei (MN) and chromosome aberration (CA) assays and determination of 8-oxo-2-deoxyguanosine (8-OH-dG) levels. The results of MTT and LDH assays showed that the higher concentrations of magnetite (100, 150, 300, 500 and 1000 mg/L) decreased cell viability. Concentrations of magnetite higher than 10 mg/L increased TOS levels and decreased TAC levels in human blood cells. Increasing concentrations of magnetite caused significant increases in MN, SCE and CA rates and 8-OH-dG levels. The obtained results showed that magnetite exerted dose-dependent effects on oxidative damage, genotoxicity and cytotoxicity in human blood cells

Synthesis, Biological Activity Evaluation and Molecular Docking of Imidazole Derivatives Possessing Hydrazone Moiety

In an attempt to identify potential active anticancer agents with low cytotoxic properties and CA inhibitors, a new series of hybrid compounds incorporating imidazole ring and hydrazone moiety as part of their structure were synthesized by aza-Michael addition reaction followed by intramolecular cyclization. The structure of synthesized compounds was elucidated using various spectral techniques. Synthesized compounds were evaluated for their in vitro anticancer (prostate cell lines; PC3) and CA inhibitory (hCA I and hCA II) activity. Among them, some compound displayed remarkable anticancer activity and CA inhibitory activity with Ki values in range of 17.53±7.19–150.50±68.87 nM against cytosolic hCA I isoform associated with epilepsy, and 28.82±14.26–153.27±55.80 nM against dominant cytosolic hCA II isoforms associated with glaucoma. Furthermore, the theoretical parameters of the bioactive molecules were calculated to establish their drug-likeness qualities. The proteins used for the calculations are prostate cancer protein (PDB ID: 3RUK and 6XXP). ADME/T analysis was carried out to examine the drug properties of the studied molecules

Djelovanje taurina protiv citogenetičkog i oksidativnog oštećenja u kulturama ljudskih limfocita uzrokovanih permetrinom

Permethrin (PM) is a common pyrethroid pesticide used to control pests in agriculture, forestry, horticulture, health care, homes, and textile industry. It is confi rmed as a strong mutagen in animals and humans. Taurine (TA) is an amino acid found in mammalian tissues that protects the cell against DNA damage. In this study, we investigated whether supplementation of human lymphocyte cultures with TA (in the concentrations of 25 μg mL-1, 50 μg mL-1 and 100 μg mL-1) provided any protection against PM toxicity applied in the concentration of 200 μg mL-1. Genotoxicity was assessed using the micronucleus (MN) and sister chromatid exchanges (SCE) tests. In addition, we measured the total antioxidant capacity (TAC) and total oxidative stress (TOS) levels in the plasma to determine oxidative effects. PM increased SCE and MN levels and altered TAC and TOS levels. TA alone did not affect SCE and MN levels compared to controls, regardless of the concentration applied. In addition, it increased TAC levels without changing TOS levels. Moreover, it signifi cantly buffered the negative cytogenetic and oxidative effects induced by PM in a clear dose-dependent manner. In conclusion, this study is the first to evidence the beneficial effects of TA against PM-induced DNA and oxidative damages in vitro.Permetrin je piretroidni pesticid koji se često rabi za suzbijanje nametnika u poljoprivredi, šumarstvu, povrtlarstvu, zdravstvenoj zaštiti, domovima i tekstilnoj industriji. Poznat je kao snažan mutagen u životinja i ljudi. Taurin je aminokiselina koja se nalazi u tkivu sisavaca i štiti stanicu od oštećenja DNA. Svrha je ovog istraživanja bila saznati hoće li taurin (u koncentracijama od 25 μg mL-1, 50 μg mL-1 te 100 μg mL-1) zaštititi ljudske limfocite od toksičnoga djelovanja permetrina koji je dodan kulturama u koncentraciji od 200 μg mL-1. Genotoksično djelovanje ocijenili smo s pomoću mikronukleus (MN)-testa i testa izmjena sestrinskih kromatida (engl. sister chromatid exchanges, krat. SCE). Da utvrdimo oksidativno djelovanje, izmjerili smo ukupni antioksidativni kapacitet (engl. total antioxidant capacity, krat. TAC) i ukupni oksidativni stres (engl. total oxidative stress, krat. TOS) u plazmi. Permetrin je povećao učestalost SCE i MN te promijenio TAC i TOS. Sam taurin, bez obzira na koncentraciju, nije utjecao na učestalost SCE i MN u odnosu prema kontroli. Usto je podigao TAC, a da pritom nije utjecao na TOS. Štoviše, značajno je ublažio štetno citogenetičko i oksidativno djelovanje permetrina, a učinkovitost mu je bila izravno povezana s primijenjenom koncentracijom. Ovo je prvo in vitro istraživanje koje je pokazalo povoljno djelovanje taurina protiv oksidacijskoga djelovanja permetrina i oštećenja DNA koje on uzrokuje