Naslov
Immunoelectronmicroscopy of the HNE-DNA adducts in normal and HNE-treated spleen

Autori
Živković, Morana ; Lunec, Joe ; Žarković, Kamelija ; Mistry, Nalini ; Žarković, Neven

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
2nd Meeting of the HNE-Club "HNE and Lipid Peroxidation Products : from basic science to medicine" / - , 2004, P44-P44

Skup
Meeting of the HNE-Club (2 ; 2004)

Mjesto i datum
Berlin, Njemačka, 06.07.2004. - 09.07.2004

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
oxidative stress; immunogold method; HNE-DNA

Sažetak
Oxidative stress is considered an important part of different disorders (cancer, cardiovascular, metabolic, degenerative and autoimmune diseases, etc.), as well as in physiological processes (in particular inflammation). Overproduction of ROS is cytotoxic, damages macromolecules and can lead to occurrence of lipid peroxidation “ end products” , among which is 4-hydroxy-2-nonenal (HNE), denoted as a “ second toxic messenger of free radicals” . HNE is considered to be mutagenic and carcinogenic on one side, but on the other it is a signaling molecule, regulating cellular proliferation, differentiation and apoptosis. It seems that most if biomedical effects of HNE are results of the aldehyde’ s affinity to bind to bioactive macromolecules. Although it is generated in excess during acute and chronic oxidative stress, HNE is also found in normal tissues under physiological circumstances. Therefore, studies on the presence of possible HNE-DNA adducts under pathological and physiological circumstances in tissue are of importance both for research and possible clinical applications. The aim of this study was to determine distribution of HNE-DNA conjugates on subcellular level. For this purpose spleen tissue on normal healthy rats was used. As immunopositive controls we used 100  M HNE treated tissues samples. Specimens were anaylsed by genuine immunogold procedure using genuine polyclonal antibodies against HNE-DNA adducts. This polyclonal antibody was raised in rabbits using as an immunogen DNA reacted with a mixture of aldehydes generated by the lipid peroxidation process. These included HNE, acrolein glyoxal and MDA (malondialdehyde) and greatest reactivity of antibodies was detected against HNE-DNA. We found HNE present in normal rat spleens very rarely and it was present almost exclusively in nucleus. In case of some cells, strong immunopositivity was observed in condensed nuclear chromatin. In case of tissue pretreatment with HNE, we detected immunopositivity spread diffusely, but again most of HNE immunopositivity was associated with nucleus, in particular chromatin. In several cells obvious immunopositivity was seen in mitochondria. Therefore, our preliminary data suggest that the novel polyclonal antiserum cold be used to detect HNE-DNA adducts by immuno-electronmicroscopy and that HNE-DNA adducts are formed even under physiological circumstances in rat spleen. Further studies are on the way.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti, Kliničke medicinske znanosti

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