Utjecaj stresa endoplazmatskog retikuluma na ekspresiju citokina IL-1beta, IL-6, IL-10 i TGF-beta u jetri miša

master's thesis

• Title (english): The inffluence of endoplasmic-reticulum stress on cytokine IL-1beta, IL-6, IL-10 and TGF-beta expression in mouse liver
• Author: Mima Bašić
• Mentor: Mirela Baus Lončar (mentor)
• Mentor: Ana Galov (mentor)
• Committee member: Ana Galov (predsjednik povjerenstva)
• Committee member: Renata Matoničkin Kepčija (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2019-09-12, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Stres endoplazmatskog retikuluma (ER) stanje je narušene homeostaze, okarakteriziran nakupljanjem nesmotanih ili pogrešno smotanih proteina. Tada se pokreće adaptivni mehanizam stanice nazvan odgovor nesmotanih proteina koji sekod sisavaca sastoji od tri glavna signalna puta. Njihova aktivacija rezultira promjenama na transkripcijskoj i translacijskoj razini u svrhu uspostavljanja normalne funkcije ER-a. Kada je stres perzistentan i nepopravljiv, odgovor nesmotanih proteina pokreće apoptozu stanice.Odgovor nesmotanih proteina može aktivirati ključni transkripcijski faktor u modulaciji upalnih odgovora, NF-κB, povećati ekspresiju proupalnih citokina i time upalne procese. Danas se intenzivno istražuje uloga stresa ER-a u patogenezi raznih bolesti. Kako bi se ustanovilo na koji način stres ER-a mijenja ekspresiju gena IL1B, IL6, IL10 i TGFB u tkivu jetre miševa, upoređena je njihova relativna ekspresija u uvjetima stresa ER-a, kod miševa tretiranih tunikamicinom, s relativnom ekspresijom u normalnim fiziološkim uvjetima u kontrolnoj grupi miševa. U tu svrhu izolirana je RNA iz tkiva jetre miševa, reverznom transkripcijom prevedena u cDNA te je analizirana metodom lančane reakcije polimerazom u stvarnom vremenu (qPCR). Analizom rezultata metode qPCR računalnim programom REST ustanovljeno je da stres ER-a uzrokuje statistički značajno povećanje ekspresije IL1B i IL6 te smanjenje ekspresije protuupalnog IL10 u jetri miša.

Abstract (english)

Endoplasmic-reticulum stress is a state of disrupted ER homeostasis, characterized by accumulation of unfolded or misfolded proteins. ER activates adaptive mechanism of cells called unfolded protein response which is comprised of three major signal pathways in mammals. Their activation results in changes on transcriptional and translational level to restore normal ER functions. In case of persistent and irremediable ER stress, unfolded protein response initiates cell apoptosis. Unfolded protein response can activate crucial transcription factor of inflammation, NF-κB, and increase expression of proinflammatory cytokines and processes. Roles of ER stress in pathology of different diseases are being intensively investigated today. To determine how ER stress changes expression of IL1B, IL6, IL10 and TGFB genes in mouse liver, their expression under conditions of ER stress in mice treated with tunicamycin was compared to their expression in normal physiological conditions in control group of mice. For that purpose RNA from mouse liver tissue was isolated, transcribed by reverse transcription into cDNA and analyzed by quantitative polymerase chain reaction (qPCR). Results obtained by qPCR were analysed by REST software. The results showed that ER stress causes statistically significant increase of IL1B and IL6 expression and statistically significant decrease of IL10 expression.

• Keywords: stres endoplazmatskog retikuluma; UPR; upala; jetra; qPCR
• Keywords (english): endoplasmic-reticulum stress; UPR; inflammation; liver; qPCR
• Language: croatian
• URN:NBN: urn:nbn:hr:217:685671
• 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
• Created on: 2019-11-29 10:08:30