Kvantitativno određivanje glutationa u tkivima štakora

master's thesis

• Title (english): Quantitive determination of gluthatione in rat tissues
• Author: Svjetlana Kovačević
• Mentor: Branimir Šimić (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Toxicology); Zagreb
• Defense date and country: 2007-05-11, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Food Technology

Abstract

Glutation igra nekoliko vrlo važnih uloga u organizmu od kojih su najznačajnije detoksikacija ksenobiotika, antioksidativno djelovanje, održavanje redoks sustava u stanici i regulacija brojnih drugih metaboličkih procesa. U stanicama se javlja u reduciranom i oksidiranom obliku od kojih je reducirani oblik zastupljen sa više od 99% od ukupne količine GSH (GSH + GSSG). U ovome radu koristila se neenzimska kolorimetrijska metoda s Ellmanovim reagensom za kvantitativno određivanje GSH u jetri, bubrezima i mozgu mužjaka štakora s naglaskom na preciznost metode i uvjete pohranjivanja uzoraka za vrijeme provođenja eksperimenta. Izmjerene količine GSH u svježim uzorcima bile su u jetri 6,8 ± 0,2 nmol/mg tkiva, u bubregu 2,9 ± 0,09 nmol/mg tkiva i u mozgu 1,5 ±0,08 nmol/mg tkiva. Postoje indicije da nakon uklanjanja organa iz organizma može doći do degradacije GSH što može dati lažne rezultate mjerenja. Dobiveni rezultati pokazali su da je kod bubrega nakon 7 dana čuvanja na -20ºC došlo do najvećeg pada količine GSH od 28,2%, a do najmanjeg pada od svega 6,7% došlo je kod mozga. Kod jetre je došlo do pada od 11,2% što je značajno ukoliko se uzorci žele pohranjivati duže vrijeme. Rezultati mjerenja količina GSH u uzorcima čuvanim 7 dana na -70 ºC pokazali su da je kod jetre došlo do pada od svega 2,3%, a kod bubrega svega 2,6%. Količina GSH u mozgu je i pri -20 ºC pokazivala malu promjenu.

Abstract (english)

Glutathione plays a couple of essential roles in the organism including detoxification of xenobiotics, antioxidative protection, maintaining redox capacity in the cell and regulation of many important metabolic processes. It is normally found in cells in reduced or oxidized form of which the reduced one is present in more than 99% of total GSH (GSH + GSSG). In this work a nonenzymatic colorimetric method with Ellmans reagent was used for quantitative determination of GSH in male rat liver, kidney and brain. The precision of the applied method and the conditions of samples storage during the experiment was followed because of possible GSH degradation after the organ removal which can cause false results of the measurements. Measured quantities of GSH in fresh samples of liver were 6.8 ± 0.2 nmol/mg tissue, in kidney were 2.9 ± 0.09 nmol/mg tissue and in the brain were 1.5 ±0.08 nmol/mg tissue. The results have shown that after 7 days of storage at -20 ºC the greatest decrease of GSH content of over than 28.2% was found in kidney and the least decrease of just 6.7% was found in the brain. In the liver, the loss was approximately 11.2% which is significant if the long-term storage of samples is necessary. The results of measuring GSH content after 7 days of storage at -70 ºC show that the loss of GSH in liver was 2.3%, and in kidney only 2.6%. The content of GSH in the brain was not significantly changed during the experiments at -20 ºC.

• Keywords: glutation; štakor; jetra; bubreg; mozak
• Keywords (english): glutathione; rat; liver; kidney; brain
• Language: croatian
• URN:NBN: urn:nbn:hr:159:597457
• Study programme: Title: Food Technology; specializations in: Nutrition; Course: Nutrition; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer prehrambene tehnologije; smjer nutricionizam (diplomirani inženjer prehrambene tehnologije; smjer nutricionizam)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-06 12:37:54