Promjene prisutnosti i raspodjele obje podjedinice feritina u nefrotoksičnisti uzrokovanoj cisplatinom

master's thesis

• Title (english): Changes in the presence and distribution of both ferritin subunits in cisplatin nephrotoxicity
• Author: Valentina Dolić
• Mentor: Marija Ljubojević (mentor)
• Mentor: Domagoj Đikić (komentor)
• Committee member: Domagoj Đikić (predsjednik povjerenstva)
• Committee member: Perica Mustafić (član povjerenstva)
• Committee member: Jasna Lajtner (član povjerenstva)
• Committee member: Sofia Ana Blažević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-09-18, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Cisplatina, planarni platinski kompleks, uspješan je kemoterapeutik za liječenje solidnih tumora. Vezanjem cisplatine za DNA dolazi do smrti neoplazmi, ali je primjena ograničena nefrotoksičnošću koja rezultira smanjenjem ili gubitkom funkcije bubrega. Od brojnih mehanizama koji doprinose nefrotoksičnosti, oksidativni stres je među izraženijim. Željezo je esencijalni element koji bubrezi nakupljaju kroz vrijeme jer se ono ne izlučuje. Time je pojačana pohrana željeza u nanokavezima feritina u proksimalnim kanalićima nefrona te opasnost od oslobađanja pri staničnom stresu. Nedovoljno je istraženo kako se ekspresija feritina mijenja tretmanom cisplatinom. Najviše oštećenja dožive stanice S3 odsječka (pars recta) proksimalnih kanalića nefrona u eksperimentalnih životinja. Ovim radom cilj nam je utvrditi dolazi li do promjene prisutnosti i raspodjele obje podjedinice feritina u nefrotoksičnosti uzrokovanoj cisplatinom. Nakon jednokratnog tretmana cisplatinom (i.p. u dozi 5 mg/kg) nefrotoksični učinak u muških štakora soja Wistar izražen je 5. dan. S pomoću specifičnih protutijela pratili smo promjene prisutnosti lakog i teškog lanca feritina, koji čine nanokavez, u arhivskim uzorcima homogenata tkiva bubrega western blot metodom. Imunohistokemijskom analizom pratili smo njihovu promjenu u raspodjeli na tkivnim kriosekcijama bubrega. Promjenu u ekspresiji nukleinskih kiselina utvrdili smo end-point RT-PCR tehnikom. Nađeno je povećanje prisutnosti mRNA lakog te proteina lakog i teškog lanca feritina te povećanje i promjena raspodjele koja ukazuje narušen metabolizam feritina kao moguć uzrok oksidativnog stresa. Proces vjerojatno kreće od feritinofagije, razgradnje u lizosomima, preko oslobađanja željeza iz feritina do iterativnog nastanka hidroksid radikala, pa dalje u feroptozu. Kontrola navedenog, uz pozitivan utjecaj na terapijsku funkciju, poželjan je ishod ovakvog saznanja.

Abstract (english)

Cisplatin, platinum cordinated planar complex, is successful chemotherapeutic, used for the treatment of solid tumors. Binding of cisplatin to DNA results in the death of neoplasms, but application is limited by nephrotoxicity resulting in reduction or loss of kidney function. Of the numerous mechanisms that contribute to nephrotoxicity, oxidative stress is among the most pronounced. Iron is an essential element that the kidneys accumulate over time because it is not excreted. This increases the storage of iron in ferritin nanocages in the proximal tubules of the nephron and the risk of its release during cellular stress. How ferritin expression is altered by cisplatin treatment has been insufficiently investigated. The cells of the S3 section (pars recta) of the proximal tubules of the nephron in experimental animals experience the most damage. The aim of this work is to determine whether there is a change in the presence and distribution of both ferritin subunits in cisplatin-induced nephrotoxicity. After a single treatment with cisplatin (i.p. at a dose of 5 mg/kg), the nephrotoxic effect in male Wistar rats was expressed on the 5th day. Using specific antibodies, we monitored changes in the presence of light and heavy ferritin chains, which make up the nanocage, in archival samples of kidney tissue homogenates by western blot analysis. Using immunohistochemical analysis, we monitored their change in distribution on kidney tissue cryosections. We determined the change in the expression of nucleic acids using the end-point RT-PCR technique. An increase in the presence of light mRNA and light and heavy ferritin protein was found, as well as an increase and change in distribution, which indicates impaired ferritin metabolism as a possible cause of oxidative stress. The process probably starts from ferritinophagy, degradation in lysosomes, through the release of iron from ferritin to the iterative formation of hydroxide radicals, and further into ferroptosis. Control of the above, with a positive impact on the therapeutic function, is the desired outcome of this kind of results.

• Keywords: autofagocitoza; feritinofagija; lizosomi; prijelazni metali; nanokavez
• Keywords (english): autophagocytosis; ferritinophagy; lysosomes; transition metals; nanocage
• Language: croatian
• URN:NBN: urn:nbn:hr:217:269937
• Project: Number: 533-03-23-0006; Title: Procjena učinkovitosti i toksičnosti biološki aktivnih tvari; Title: Assessment of efficacy and toxicity of biologically active substances; Acronym: BioMolTox; Leader: Nikolina Maček Hrvat; Jurisdiction: eu; Funder: Europska unija
• Study programme: Title: Experimental Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra eksperimentalne biologije (sveučilišni/a magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-10-03 10:02:29