Naslov
In vitro model of intestinal barrier in oxidative stress research

Autori
Štroser, Marina ; Borović Šunjić, Suzana ; Živković, Emilija ; Stolc, Svorad ; Žarković, Neven

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

Izvornik
Herman Esterbauer Meeting From Biochemistry to Human Disease : Reactive Oxygen Species & Antioxidants : book of abstracts / - , 2007

Skup
Herman Esterbauer Meeting From Biochemistry to Human Disease : Reactive Oxygen Species & Antioxidants

Mjesto i datum
Graz, Austrija, 15.10.2007. - 18.10.2007

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
CaCo-2; differentiation; 4-hydroxynonenal; HNE; intestinal barrier; antioxidants; oxidative stress; gastrointestinal tract

Sažetak
Human colon adenocarcinoma cell line CaCo-2 was shown to undergo spontaneous in vitro enterocytic differentiation leading to the formation of cell monolayer with several morphological an functional characteristics of mature enterocytes. Gastrointestinal tract is often exposed to harmful compounds originating from diet ; they can cause oxidative stress and intestinal dysfunction. Such dysfunction can make the intestinal barrier permeability greater for toxins, bacteries, harmful products of digestion, free radicals and second messangers of free radicals like 4-hydroxynonenal (HNE). HNE is one of major lipid peroxidation product, and because of its lipophilic caracter, it can be readily absorbed in the intestine or can accumulate in membranes causing phospholipid and protein damage. Aims of this study were: establishment of intestinal barrier model by growing CaCo-2 cells ; exploring the effect of HNE on this model ; and determining effects of several antioxidants in preventing damaging effects of HNE. Toxicity of HNE and antioxidants used in experiment was determined by MTT assay and cell viabilty by Tripan blue staining. CaCo-2 cells were grown in cell culture inserts with 0, 4 µ ; m pores.. Differentiation of the monolayer was determined by measuring activity of alkaline phosphatase and reading apsorbance at 405 nm on plate reader. Permeability of the differentiated monolayer was measured by the difusion of phenol red solution (2 mg/ml) prepared in Hank's buffer through cell monolayer into plate well with Hank's buffer without color. Percentage of difusion was presented as ratio of measured apsorbances at 556 nm before and after treatment. Cell monolayer is good developed after 15th day and represents intestinal barrier model. HNE impaired function of intestinal barrier model and caused increase in its permeability when used as 160 µ ; M. N-acetylcystein (NAC) showed the best effect since it managed to completely protect cells from damaging effects of HNE ; permeability was blocked and integrity of cell monolayer was preserved. With this study, we have shown that 15 days is enough time for cells to differentiate and develop consistent monolayer that represents intestinal barrier and good model for oxidative stress research. HNE impaired function of intestinal barrier model, while NAC completely prevented this

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA