diplomsko delo univerzitetnega študijskega programa I. stopnje
Tjaša Brenčič (Author), Uroš Potočnik (Mentor), Vojko Berce (Co-mentor)

Abstract

Astma je kompleksna genetska bolezen s heterogenim fenotipom, ki ni posledica samo posameznih genetskih ali samo okoljskih dejavnikov, ampak prihaja do nje z mešanjem obeh. V diplomski nalogi smo se osredotočili na vlogo endokanabinoidnega sistema v imunskem sistemu, ki sodeluje pri regulaciji številnih fizioloških in patoloških funkcij, prav tako pa je prisoten pri protivnetnem odgovoru imunskega sistema ob različnih boleznih, zato bi lahko bil potencialna tarča za zdravljenje astme. V diplomski nalogi smo primerjali gensko ekspresijo kanabinoidnih receptorjev CNR1 in CNR2 med skupino bolnikov z astmo in kontrolno skupino ter genotipizirali štiri polimorfizme posameznega nukleotida (SNP), za katere smo predvidevali, da bodo povezani z izbranima genoma. Ugotavljali smo, kakšen vpliv imajo izbrani polimorfizmi na klinične parametre pred zdravljenjem z glukokortikoidi in antilevkotrieni in po njem, ter jih primerjali z gensko ekspresijo le-teh. Ekspresijo genov CNR1 in CNR2 smo merili z reakcijo PCR v realnem času v krvnih limfocitih 340 otrok, obolelih za astmo, in 257 zdravih otrok, starih med 5 in 18 let. Bolnike smo razdelili v dve skupini glede na fenotip (alergijsko in nealergijsko obliko). Za genotipizacijo smo uporabili tehniko analize talilne krivulje visoke ločljivosti (HRM). Rezultate smo statistično obdelali s programskim paketom GraphPad Prism 6. Ugotovili smo kar nekaj povezav med polimorfizmi SNP rs4237, rs2229579 in rs2501432 ter genom CNR2, tako s kliničnimi parametri kot z gensko ekspresijo. Polimorfizma SNP rs13197090 nismo povezali z genom CNR1, smo pa ugotovili, da je pri bolnikih z astmo izražanje tega gena izrazito povečano. Tako smo prvi, ki smo ugotovili, da je ekspresija obeh genov izrazito povečana pri bolnikih z astmo in da izbrani polimorfizmi za gen CNR2 vplivajo na delovanje le-tega. Iz dobljenih rezultatov je mogoče sklepati, da imata gena CNR1 in CNR2 pomembno vlogo pri patogenezi astme zato bi lahko v bodoče služila kot biomarkerja v medicinski diagnostiki ter pripomogla h kvalitetnejšemu zdravljenju bolnikov z astmo.

Keywords

astma;asociacijska študija;polimorfizem posameznega nukleotida;genotipizacija;genska ekpresija;kanabidni receptor;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher: [T. Brenčič]
UDC: 577.171.6(043.2)
COBISS: 19432982 Link will open in a new window
Views: 1748
Downloads: 150
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: GENOTYPING AND EXPRESSION OF CANNABINOID RECEPTORS CNR1 AND CNR2 IN ASTHMA PATIENTS
Secondary abstract: Asthma is a complex genetic disease with heterogeneous phenotype which is not only the consequence of individual genetic or environmental factors but the result of both. In the diploma thesis we focused on the role of endocanabinoid system in the immune system which participates in regulation of many physiological and pathological functions. It is also present as a anti-inflammatory answer of the immune system in different diseases. Therefore it could be a potential target for treatment of asthma. The aim of our study was to compare gene expression of canabinoids CNR1 and CNR2 receptors between the group of asthma patients and the control group. We also genotyped four polymorphisms SNP for which we predicted that they are connect with our two genes. Further we compared the expression of CNR1 and CNR2 receptor with clinical parameters before and after anti-asthmatic treatment. We measured gene expression of CNR2 receptor with real-time PCR in blood lymphocytes of 340 children with asthma and 257 healthy children between the ages 5 and 18 of the Slovenian population. The patients had been analysed according to the phenotype (allergic and non allergic group). Polymorphisms had been genotyped with HRM analysis. Results had been analysed with statistic program GraphPad Prism 6. The results confirmed our hypothesis that the expression of canabinoids CNR2 receptor is higher in astma patients. We determined that polymorphisms SNP rs4237, SNP rs2229579, SNP rs2501432 are associated with genotype, phenotype and clinical parameters before and after the treatment and the expression of CNR2 receptor. We did not connect polymorphism SNP rs13197090 with gene CNR1. We were the first one who associated receptors CNR1 and CNR2 with asthma and determined that they are strongly up-regulated in asthma patients. From that we can conclude that gene CNR1 and CNR2 has an important role in patogenesis of asthma and in the future it could serve as a biomarkers in medical diagnostics and contribute to the quality of treatment of asthma patients.
Secondary keywords: asthma;asociation study;single nucleotide polymorphism;genotyping;gene expression;cannabinoid receptors;
URN: URN:SI:UM:
Type (COBISS): Bachelor thesis/paper
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: XI, 81 str.
ID: 8887472