20.500.12556/RUL-186941 Vpliv 1,25-dihidroksivitamina D₃ na z LPS sprožen vnetni odziv primarnih epitelnih celic goveje mlečne žleze Effect of 1,25-dihydroxyvitamin D₃ on the LPS-induced inflammatory response in primary bovine mammary epithelial cells Mastitis pri govedu predstavlja velik zdravstveni in ekonomski iziv v prireji mleka, zato se raziskave zaradi naraščajoče antibiotične rezistence usmerjajo v proučevanje prirojene imunosti ter iskanju alternativnih metod nadzora. Imunski odziv v vimenu temelji na mlečnih celicah ter imunskih celicah, ki preko receptorjev TLR prepoznavajo bakterijske molekularne vzorce, kot je LPS. Aktivacija signalnih poti vodi do sproščanja vnetnih citokinov (TNF α, IL-1β, IL-6) ter kemokina IL-8, kar povzroči rekrutacijo nevtrofilcev na mesto okužbe in povečanje števila somatskih celic v mleku. Za preučevanje teh mehanizmov in vitro se pogosto uporabljajo primarne goveje mlečne epitelne celice. Ker lahko prekomerno vnetje poškoduje žlezno tkivo, je pomembna imunomodultorna vloga vitamina D (1,25(OH)₂D₃), ki preko receptorja VDR spodbuja sintezo protimikrobnih peptidov in umirja vnetni odziv. V okviru eksperimentalnega dela smo na pbMEC preučevali vpliv predhodnega tretiranja z vitaminom D pri dveh različnih koncentracijah (10 nM in 100 nM) na z LPS sprožen vnetni odziv. Z metodo qPCR smo analizirali spremembo v diferenčnem izražanju vnetnih genov (TNF-α, IL-8, NF-κB, MyD88, IL-6) in primerjali zgodnji (3 ure pred izolacijo RNA) in kasnejši (24 ur pred izolacijo RNA) imunski odziv. Rezultati so potrdili, da dodatek LPS sproži vnetni odziv, predhodno tretiranje z vitaminom D pa je pokazalo potencialni imunomodulatorni učinek. Pokazali smo tudi celično specifičnost imunskega odziva, medtem ko neposredne odvisnosti od koncentracije vitamina D nismo potrdili. Bovine mastitis represents a major health and economic challenge in dairy production. Due to rising antibiotic resistance, research is increasingly focusing on understanding innate immunity and searching for alternative approaches for disease control. The immune response in the udder relies on mammary epithelial cells and immune cells, which recognize pathogen-associated molecular patterns, such as lipopolysaccharide (LPS), via TLR receptors. Activation of downstream signalling pathways triggers the release of inflammatory cytokines (TNF-α, IL-1β, IL-6) and the chemokine IL-8, leading to the recruitment of neutrophils to the site of infection and an increase in the somatic cell count in milk. Primary bovine mammary epithelial cells are widely used as in vitro models to study these mechanisms. Because excessive inflammation can cause significant damage to the glandular tissue, the immunomodulatory role of vitamin D (1,25(OH)₂D₃) is crucial, as it stimulates the synthesis of antimicrobial peptides and dampens the inflammatory response via the VDR receptor. In the experimental part of this study, we evaluated the effect of pretreatment with vitamin D at two different concentrations (10 nM and 100 nM) on the LPS induced inflammatory response in pbMEC. Using qPCR, we analysed changes in the differential gene expression of inflammatory genes (TNF α, IL 8, NF κB, MyD88, IL 6) and compared early (3 hours prior to RNA isolation) and late (24 hours prior to RNA isolation) immune responses. The results confirmed that LPS stimulation triggers an inflammatory response, whereas pretreatment with vitamin D demonstrated a potential immunomodulatory effect. We also demonstrated cell line specificity of the immune response, although a direct dependency on vitamin D concentration was not confirmed. mastitis epitelne celice mlečne žleze vnetje vitamin D imunski odziv mastitis mammary epithelial cells inflammation vitamin D immune response true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-09-07 13:05:04 2026-09-07 13:05:15 2026-09-08 04:19:42 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 25955 Ciglar_Nika_-_Vpliv_1,25-dihidroksivitamina_D₃_na_z_LPS_sprozen_vnetni_odziv_primarnih_epitelnih.pdf Ciglar_Nika_-_Vpliv_1,25-dihidroksivitamina_D₃_na_z_LPS_sprozen_vnetni_odziv_primarnih_epitelnih.pdf 1 9A2E7A521BAAC07DC558677187A02D41 5f5caf445d59e4ebce844cc7ebc8b689ed88df354aefb1cf53b6da9fd83fc073 fdc0c6c0-aaab-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=246341 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0