T-2 toxin is a worldwide trichothecenetoxin and can cause various toxicities.T-2 toxin is involved in G1 phase arrest in several cell lines but molecular mechanism is still not clear. In present study, we used rat pituitary GH3 cells to investigate the mechanism involved in cell cycle arrest against T-2 toxin (40 nM) for 12, 24, 36 and 48 h as compared to control cells. GH3 cells showed a considerable increase in reactive oxygen species (ROS) as well as loss in mitochondrial membrane potential (△Ym) upon exposure to the T-2 toxin. Flow cytometry showed a significant time-dependent increase in percentage of apoptotic cells and gel electrophoresis showed the hallmark of apoptosis oligonucleosomal DNA fragmentation. Additionally, T-2 toxin-induced oxidative stress and DNA damage with a time-dependent significant increased expression of p53 favors the apoptotic process by the activation of caspase-3 in T-2 toxin treated cells. Cell cycle analysis by flow cytometry revealed a time-dependent increase ofG1 cell population along with the significant time-dependent up-regulation of mRNA and protein expression of p16 and p21 and significant down-regulation of cyclin D1, CDK4, and p-RB levels further verify the G1 phase arrest in GH3 cells. Morphology of GH3 cells by TEM clearly showed the damage and dysfunction to mitochondria and the cell nucleus. These findings for the first time demonstrate that T-2 toxin induces G1 phase cell cycle arrest by the involvement of p16/Rb pathway, along with ROS mediated oxidative stress and DNA damage with p53 and caspase cascade interaction, resulting in apoptosis in GH3 cells.
- MeSH
- buněčné linie MeSH
- buněčný cyklus účinky léků fyziologie MeSH
- geny p16 účinky léků fyziologie MeSH
- hypofýza účinky léků metabolismus ultrastruktura MeSH
- krysa rodu rattus MeSH
- retinoblastomový protein biosyntéza MeSH
- signální transdukce účinky léků fyziologie MeSH
- T-2 toxin toxicita MeSH
- viabilita buněk účinky léků fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
T-2 toxin, a trichothecene mycotoxin, is a common contaminant in food and animal feed, and is also present in processed cereal products. The most common route of T-2 toxin exposure in humans is through dietary ingestion. The cytotoxic effects of T-2 toxin include modifications to feeding behavior, nervous disorders, cardiovascular alterations, immunosuppression, and hemostatic derangements. However, to date, effects on the central nervous system (CNS) have rarely been reported. In the present study, female Wistar rat were given a single dose of T-2 toxin at 2 mg/kg b.w. and were sacrificed at one, three, and seven days post-exposure. Histopathological analysis and transmission electron microscope (TEM) observations were used to investigate injury to the brain and pituitary gland. Damage to the brain and pituitary at the molecular level was detected by real time-polymerase chain reaction (RT-PCR), western blot, and immunohistochemical assays. Liquid chromatograph-mass spectrometer/mass spectrometer (LC-MS/MS) was used to investigate T-2 concentration in the brain. The results showed that pathological lesions were obvious in the brain at three days post-exposure; lesions in the pituitary were not observed until seven days post-exposure. Autophagy in the brain and apoptosis in the pituitary suggest that T-2 toxin may induce different acute reactions in different tissues. Importantly, low concentrations of T-2 toxin in the brain were observed in only one rat. Responsible for the above mentioned, we hypothesize that brain damage caused by this toxin may be due to the ability of the toxin to directly cross the blood-brain barrier (BBB). Therefore, given its widespread pollution in food, we should pay more attention to the neurotoxic effects of the T-2 toxin, which may have widespread implications for human health.
- MeSH
- apoptóza účinky léků MeSH
- autofagie účinky léků MeSH
- časové faktory MeSH
- chování zvířat účinky léků MeSH
- chromatografie kapalinová MeSH
- hematoencefalická bariéra metabolismus MeSH
- hodnocení rizik MeSH
- hypofýza účinky léků metabolismus ultrastruktura MeSH
- imunohistochemie MeSH
- kapilární permeabilita MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- mozek účinky léků metabolismus ultrastruktura MeSH
- neurotoxické syndromy etiologie metabolismus patologie psychologie MeSH
- potkani Wistar MeSH
- regulace genové exprese MeSH
- T-2 toxin metabolismus toxicita MeSH
- tandemová hmotnostní spektrometrie MeSH
- transmisní elektronová mikroskopie MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- histocytochemie MeSH
- hypofýza metabolismus ultrastruktura MeSH
- Perciformes anatomie a histologie metabolismus MeSH
- roční období MeSH
- rozmnožování * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- embryo savčí MeSH
- hypofýza ultrastruktura MeSH
- krysa rodu rattus MeSH
- Check Tag
- krysa rodu rattus MeSH
