Učinak stresa endoplazmatskog retikula na ekspresiju gena TFF3, TNFalfa, MCP1 i CXCL1 u jetri miša

master's thesis

• Title (english): The effect of endoplasmic reticulum stress on the gene expression of Tff3, TNFalfa, MCP1 and CXCL1 in mice liver
• Author: Tajana Burkuš
• Mentor: Mirela Baus Lončar (mentor)
• Mentor: Julija Erhardt (sumentor)
• Committee member: Julija Erhardt (predsjednik povjerenstva)
• Committee member: Nenad Malenica (član povjerenstva)
• Committee member: Tvrtko Dražina (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2020-09-29, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Stres endoplazmatskog retikula (ER) nastaje nakupljanjem krivo namotanih ili nenamotanih proteina u lumenu endoplazmatskog retikula, a ponajviše u sekretornim stanicama, koje su, zbog svojih funkcija, bogate ER-om. U odgovor na stres, eukariotska stanica razvila je adaptivne mehanizme poznate kao odgovor na nenamotane proteine (UPR) čije nastojanje je vratiti staničnu homeostazu. Homeostaza se vraća zaustavljanjem translacije proteina, aktivacijom molekularnih šaperona ili degradacijom krivo namotanih proteina, a produljeni stres dovodi do patoloških promjena i stanične smrti. Hepatociti su, kao stanice s velikim kapacitetom sinteze proteina, podložni aktivaciji UPR puteva koji igraju važnu ulogu u prevenciji i patogenezi bolesti jetre. Sve više istraživanja usmjerava se prema povezanosti ER stresa, upale, stanične ozljede i steatoze u nealkoholnoj bolesti masne jetre (NAFLD) gdje glavnu ulogu imaju upalni citokini i kemokini. U ovom istraživanju razvijena je qPCR metoda (optimalni reakcijski uvjeti i početnice) za mišje tkivo jetre soja C57Bl6Crl. ER stres je izazvan jednokratnom primjenom tunikamicina, te je ispitana relativna promjena ekspresije upalnih citokina, TNFα, MCP1 i CXCL1 te proteina TFF3 čija je uloga u jetri još uvijek nedovoljno istražena. Rezultati su analizirani programom REST 2009 nakon čega je ustanovljena statistički značajna, povišena ekspresija kod Tnfα, Mcp1 i Cxcl1 gena te statistički neznačajna, smanjena ekspresija Tff3 gena.

Abstract (english)

Endoplasmic reticulum stress (ER) is characterized by the accumulation of misfolded and unfolded proteins in the ER lumen, mostly in secretory cells, which, due to their functions, are rich in the ER. In response to stress, the eukaryotic cell has evolved adaptive mechanisms known as the unfolded protein response (UPR) whose main goal is to restore cellular homeostasis. Homeostasis is restored by stopping protein translation, activation of molecular chaperones, or degradation of misfolded proteins, and can lead to cell death. Unresolved chronic ER stress leads to pathological changes. Hepatocytes have large capacity for protein synthesis and they are highly exposed to possiiblities of ER stress. Activation of UPR pathways has an important role in the prevention of liver pathogenesis. Growing body of evidence connects ER stress with inflammation, cell injury, and steatosis in nonalcoholic fatty liver disease (NAFLD) where inflammatory cytokines and chemokines play a major role. The qPCR method (optimal reaction conditions and primers) was developed for C57Bl6Crl mouse strain liver tissue. ER stress was induced with single tunicamycin injection and expression of inflammatory cytokines, TNFα, MCP1, CXCL1, and the TFF3 protein, whose role in the liver is still insufficiently investigated, was examined. The results were analyzed by the REST 2009 software. ER stress induced statistically significant increased expression of Tnfα, Mcp1, and Cxcl1 genes while expression of the Tff3 gene was not significantly changed.

• Keywords: stres endoplazmatskog retikula; jetra; Tff3; Tnfα; Mcp1; Cxcl1; qPCR
• Keywords (english): (ER stress; liver; Tff3; Tnfα; Mcp1; Cxcl1; qPCR)
• Language: croatian
• URN:NBN: urn:nbn:hr:217:477821
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-09-29
• Created on: 2020-10-07 09:12:39