Josip Juraj Strossmayer University of Osijek. Faculty of Medicine.
Abstract
Cilj istraživanja: Ekstrahirati i okarakterizirati gangliozide (lipide) stanične membrane hepatocita i proučiti njihovu ulogu u patogenezi inzulinske rezistencije kod predijabetičnih ženki štakora. Na istim životinjama procijeniti učinkovitost uobičajene antidijabetske terapije u održavanju fiziološkog omjera gangliozida u staničnim membranama. Nacrt studije: Istraživanje je provedeno kao studija parova. Materijali i metode: Prikupljeno je 27 uzoraka tkiva jetre iz 4 skupine ženki Sprague Dawley štakora hranjenih standardnom ili HFHS dijetom te tretiranih metforminom odnosno liraglutidom. Uzorci su obrađeni modificiranom metodom po Schnaaru kojom se pomoću različitih organskih otapala ekstrahiraju gangliozidi iz jetrenog tkiva. Iz svake skupine nasumično su odabrana tri uzorka. Za kvalitativnu analizu gangliozida korištena je modificirana metoda tankoslojne kromatografije po Schnaaru i Needhamu. Gangliozidi su vizualizirani prskanjem HPTLC pločice rezorcinolom, fotografirani i kvantificirani programom ImageJ. Rezultati i zaključak: Moguće je da masna i slatka hrana utječe na povećanje ekspresije jednostavne forme gangliozida GM3 i time dovodi do promjene omjera gangliozida staničnih membrana hepatocita. Antidijabetici metformin i liraglutid nemaju značajan učinak u održavanju fiziološkog omjera tih gangliozida u ženskoj populaciji.Objectives: The aim of this study was to extract and differentiate hepatocyte cell membrane gangliosides (lipids) and study their role in the pathogenesis of insulin resistance in prediabetic female rats. By observing the same animals, the effectiveness of the most frequently used antidiabetic therapy was evaluated in maintaining the physiological ganglioside ratio in cell membranes. Study design: The study was conducted as a matched pair study. Material and methods: 27 liver tissue samples were collected from 4 groups of Sprague Dawley female rats fed with regular or HFHS diet and treated with metformin or liraglutide. The samples were processed following a modified Schnaar method which involves using various organic solvents to extract gangliosides from the liver tissue. Three samples were randomly selected from each group. A thin layer chromatography method modified by Schnaar and Needham was used for the qualitative analysis of gangliosides. They were then visualized by spraying HPTLC plates with resorcinol, photographed and quantified using a computer program ImageJ. Results and conclusion: It is possible that high-fat and high-sugar diet increases the expression of a simple form of ganglioside GM3 and hence leads to a change in the hepatic cell membrane ganglioside ratio. Antidiabetic drugs metformin and liraglutide do not have a significant effect on maintaining the physiological proportion of these gangliosides in the female population
