cytochrome-c-t and deoxynivalenol

Zeolitic imidazolate framework (ZIF-8) base-aptamer "gate-lock" biomaterial probes have been synthesized for monitoring intracellular deoxynivalenol (DON) and cytochrome c (cyt c) levels. The aptamer and organic fluorescent dye were regarded as a recognition element and a sensing element, respectively. In the presence of DON, the aptamers of DON and cyt c were specifically bound with the DON and induced cyt c, leading to the dissociation of aptamers from the porous surface of the probes. The gate was subsequently opened to release methylene blue (MB) and Rhodamine 6G (Rh6G), and their fluorescence (emission of MB at 700 nm and Rh6G at 550 nm) significantly recovered within 6 h. Cell imaging successfully monitored the exposure of DON and the biological process of cyt c discharge triggered by the activation of the DON-induced apoptosis pathway. In addition, the response between DON and cyt c was observed during the apoptosis process, which is of high significance for the comprehensive and systematic development of mycotoxins cytotoxicity.

Topics: Aptamers, Nucleotide; Cytochromes c; Trichothecenes; Zeolites

Deoxynivalenol (DON) is considered a mycotoxin that is toxic to the agricultural environment and human body. It is necessary to study the pathophysiological mechanism of DON toxicity at the cellular level. Cytochrome c (Cyt c), as an important biomarker of DON-induced apoptosis that may lead to a bipartite 'point-of-no return' event, is of great significance to be detected using cell imaging. Herein, we synthesized a DON-deactivated emission fluorescent probe, the molecularly imprinted polymer-coated quantum dots (CdTe@MIP), for monitoring the Cyt c level with a photoinduced electron transfer strategy. The CdTe@MIP probe can be easily loaded into cells and perform well due to its great sensitivity and selectivity and its fluorescence was gradually quenched with the increasing concentration (0-10 μM) and incubation time (0-7.5 h) of DON. Cell imaging results of apoptosis induced by DON was consistent with that of the cell counting kit-8 assay and flow cytometry technique. The developed method can be used to monitor DON-induced apoptosis and provide an early-warning system for the contaminant toxicity.

Topics: Apoptosis; Cadmium Compounds; Cytochromes c; Electrons; Fluorescent Dyes; Humans; Quantum Dots; Tellurium; Trichothecenes

Deoxynivalenol (DON) has broad toxicity in animals and humans. In this study the impact of DON treatment on apoptotic pathways in PC12 cells was determined. The effects of DON were evaluated on (i) typical indicators of apoptosis, including cellular morphology, cell activity, lactate dehydrogenase (LDH) release, and apoptosis ratio in PC12 cells, and on (ii) the expression of key genes and proteins related to apoptosis, including Bcl-2, Bax, Bid, cytochrome C (Cyt C), apoptosis inducing factor (AIF), cleaved-Caspase9, and cleaved-Caspase3. DON treatment inhibited proliferation of PC12 cells, induced significant morphological changes and apoptosis, promoted the release of Cyt C and AIF from the mitochondria, and increased the activities of cleaved-Caspase9 and cleaved-Caspase3. Bcl-2 expression decreased with increasing DON concentrations, in contrast to Bax and Bid, which were increased with increasing DON concentration. These data demonstrate that DON induces apoptosis in PC12 cells through the mitochondrial apoptosis pathway.

Topics: Animals; Apoptosis; Caspase 3; Cytochromes c; Hazardous Substances; Mitochondria; PC12 Cells; Proto-Oncogene Proteins c-bcl-2; Rats; Trichothecenes

The Fusarium genus of fungi is responsible for commercially devastating crop diseases and the contamination of cereals with harmful mycotoxins. Fusarium mycotoxins aid infection, establishment, and spread of the fungus within the host plant. We investigated the effects of the Fusarium mycotoxin deoxynivalenol (DON) on the viability of Arabidopsis cells. Although it is known to trigger apoptosis in animal cells, DON treatment at low concentrations surprisingly did not kill these cells. On the contrary, we found that DON inhibited apoptosis-like programmed cell death (PCD) in Arabidopsis cells subjected to abiotic stress treatment in a manner independent of mitochondrial cytochrome c release. This suggested that Fusarium may utilise mycotoxins to suppress plant apoptosis-like PCD. To test this, we infected Arabidopsis cells with a wild type and a DON-minus mutant strain of F. graminearum and found that only the DON producing strain could inhibit death induced by heat treatment. These results indicate that mycotoxins may be capable of disarming plant apoptosis-like PCD and thereby suggest a novel way that some fungi can influence plant cell fate.

Topics: Apoptosis; Arabidopsis; Cell Survival; Cycloheximide; Cytochromes c; Ethanol; Fusarium; Heat-Shock Response; Hot Temperature; Mitochondria; Suspensions; Trichothecenes

Deoxynivalenol (DON), known colloquially as "vomitoxin", is a pathogenic mycotoxin produced by Fusarium fungi. Human food poisoning outbreaks, with nausea, diarrhea, and vomiting as primary symptoms, have been associated with Fusarium-infected cereals. Therefore, this study was designed to determine the molecular aspects of DON in human colon cancer cells (HT-29). To this aim, we have monitored the effects of DON on (i) cellular morphological changes via optical and transmission electron microscopy, especially in regards to cell viability and mitochondria changes, and (ii) its effects on key regulators of cell apoptosis, including cytochrome c, caspase-9, caspase-3, Bcl-2, Bax, and Bid. Our results showed that DON treatment inhibited cell proliferation, induced significant morphological changes, and promoted the activation of cytochrome c and caspases. Furthermore, changes in Bcl-2, Bax, and Bid expression were detected. The relative expression profile of Bcl-2 was contrary to that of Bax and Bid, as Bcl-2 expression decreased as the concentrations DON increased, reaching a minimum at the highest concentration of DON. We concluded that DON-induced apoptosis was caused by mitochondrial dysfunction and subsequent release of cytochrome c into the cytoplasm and successive activation of caspases, and this was likely regulated by Bcl-2 family proteins.

Topics: Apoptosis; bcl-2-Associated X Protein; BH3 Interacting Domain Death Agonist Protein; Caspases; Cell Proliferation; Colonic Neoplasms; Cytochromes c; Cytoplasm; Gene Expression Regulation, Neoplastic; HT29 Cells; Humans; Microscopy, Electron, Transmission; Mitochondria; Proto-Oncogene Proteins c-bcl-2; Trichothecenes

Deoxynivalenol (DON) is a widespread trichothecene mycotoxin which contaminates cereal crops and harmfully affects the gastrointestinal tract. Since it is well known that mitochondria play a central role in apoptosis triggered by many stimuli, an effort was made to examine whether DON-induced cytotoxicity occurs through mitochondria-mediated apoptotic pathway. The intestinal system being one of the primary targets of mycotoxins, the human colon carcinoma cell line HCT116 was used in this study. Using flow cytometric analyses and immunofluorescence, we showed that DON at 100 μM induced a mitochondria-dependent apoptotic pathway associated with opening of the mitochondrial permeability transition pore (PTP), loss of the mitochondrial transmembrane potential (ΔΨm), downstream generation of O₂·⁻ and cytochrome c release. The DON-induced apoptosis was accompanied by an activation of caspase 9 and 3, as demonstrated by Western blot and caspase activity assay. In addition, by taking advantage of HCT116 cells invalidated for Bax, we showed that this pro-apoptotic protein favored mitochondrial alterations induced by the mycotoxin. Besides, incubation of purified mitochondria with DON indicated that this mycotoxin does not directly target mitochondria to induce PTP-dependent permeabilization of mitochondrial membranes. Altogether, our results indicate that mitochondria-related caspase-dependent apoptotic pathway is involved in this in vitro model of DON induced-cytotoxicity.

Topics: Apoptosis; Caspases; Cell Line, Tumor; Cytochromes c; Enzyme Activation; Humans; Matrix Metalloproteinases; Membrane Potentials; Mitochondria; Trichothecenes