Ekspresija alfa podjedinice NaK-ATPaze u nefrotoksičnosti uzrokovanoj cisplatinom

master's thesis

• Title (english): Expression of alpha subunit of NaK-ATPase in cisplatin nephrotoxicity
• Author: Daria Sudac
• Mentor: Marija Ljubojević (mentor)
• Mentor: Domagoj Đikić (komentor)
• Committee member: Domagoj Đikić (predsjednik povjerenstva)
• Committee member: Ivana Šola (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-02-01, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Bubrezi su parni organi smješteni u trbušnoj šupljini i zaduženi za filtraciju krvi, izlučivanje
štetnih tvari i općenito održavanje homeostaze unutrašnjeg okoliša tijela. Zbog njihove
anatomije i funkcije podložni su oštećenjima uzrokovanim toksičnim spojevima uključujući
okolišne kemikalije, metale i različite lijekove. Zbog važne funkcije u reapsorbciji korisnih i
sekreciji štetnih tvari proksimalni kanalić, pogotovo njegov S3 segment, osjetljiv je na
različite ksenobiotike. Cisplatina je važan citostatik korišten za tretman različitih malignih
tumora. Jedan od glavnih nedostataka cisplatine je njezina nefrotoksičnost. Kratkoročno,
tretman cisplatine dovodi do akutnog zatajenja bubrega koje uzrokuje različite patofiziološke
promjene. Dugoročno postoji mogućnost razvoja kronične bolesti bubrega (CKD), pri kojoj se
funkcija bubrega progresivno smanjuje. Kod CKD uvijek je prisutna fibroza bubrega, za koju
je karakteristična akumulacija i aktivacija fibroblasta i miofibroblasta. Poznato je da je proces
epitelno-mezenhimske tranzicije uključen u nastanak fibroblasta iz epitelnih stanica tubula. U
ovaj proces uključena je NaK ATPaza, proteinska pumpa zadužena za aktivni transport natrija
i kalija suprotno kemijskom gradijentu. U ovom istraživanju sam pomoću metoda
imunohistokemije i western blota pokazala da dolazi do smanjenja ekspresije alfa podjedinice
NaK ATPaze na razini proteina kod štakora nakon jednokratnog tretmana cisplatinom.
Također je vidljiva povećana ekspresija mRNA za isti protein, dobivena metodom RT-PCR.

Abstract (english)

Kidneys are pair organs located in the abdominal cavity and they filter blood, excrete harmful
substances and in general maintain the homeostasis of the body’s internal environment.
Because of their anatomy and function, they are susceptible to damage caused by toxic
substances such as environmental chemicals, metals and various drugs. Due to its important
function in reabsorption of useful and secretion of harmful substances the proximal tubule,
especially in its S3 segment, kidneys are susceptible to damage by various xenobiotics.
Cisplatin is an important cytostatic used for the treatment of various malignant tumors. One of
the main disadvantages of cisplatin is its nephrotoxicity. In the short term, cisplatin treatment
leads to acute renal failure that causes various pathophysiological changes. In the long term
there is a possibility of developing chronic kidney disease (CKD), in which kidney function
progressively decreases. Kidney fibrosis is always present in CKD, and it is characterized by
the accumulation and activation of fibroblasts and myofibroblasts. It is known that the process
of epithelial-mesenchymal transition is involved in the formation of fibroblasts from tubular
epithelial cells. NaK ATPase, a protein pump responsible for the active transport of sodium
and potassium against the chemical gradient, is involved in this process. Here, using
immunohistochemistry and western blot methods, I showed that there is a decrease in the
expression of the alpha subunit of NaK ATPase at the protein level in rats, after single
treatment with cisplatin. Increased expression of mRNA for the same protein, obtained by the
RT-PCR method, is also visible.

• Keywords: proksimalni kanalić; fibroza; epitelno-mezenhimska tranzicija
• Keywords (english): proximal tubule; fibrosis; epithelial-mesenchymal transition
• Language: croatian
• URN:NBN: urn:nbn:hr:217:545515
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-04-05 10:11:51