20.500.12556/RUL-186058 Vpliv imunostimulacije primarne epitelijske celične kulture mlečne žleze goveda z lipopolisaharidom in lipoteihoično kislino na izražanje izbranih genov Effect of immunostimulation of primary bovine mammary epithelial cell cultures with lipopolysaccharide and lipoteichoic acid on the expression of selected genes Mastitis je vnetje mlečne žleze, ki prizadene skoraj polovico krav mlekaric in posledično zada velike finančne izgube mlečni industriji. Bolezen lahko močno zniža količino in kvaliteto mleka, saj vpliva na količine in razmerja med mlečnimi proteini. Mastitis povzročajo različni patogeni, najpogosteje pa sta povzročitelja E. coli in S. aureus. Želeli smo preveriti, kako lipopolisaharid (LPS) in lipoteihoična kislina (LTA), ki sta del celične stene teh patogenov ter virulentna dejavnika, vplivata na imunski odziv in sintezo proteinov v mlečni žlezi. V ta namen smo vzpostavili primarno celično kulturo epitelijskih celic mlečne žleze goveda in jo 24 ur stimulirali z virulentnima dejavnikoma. Z metodo qPCR smo preverili izražanje genov za vnetne dejavnike in ugotovili, da se po imunostimulaciji njihovo izražanje poveča, pri čemer LPS povzroči močnejši imunski odziv kot LTA. V primeru imunostimulacije z LPS-om sta se v kulturah povišali koncentraciji vnetnih citokinov IL-8 in TNF-α, medtem ko so se v primeru LTA znižali, kar smo zaznali tudi z ELISA. Vzpostavljene kulture so bile sposobne sintetizirati nekatere mlečne proteine, kar smo potrdili z imunobarvanjem. Z analizo diferenčnega izražanja genov smo ugotovili, da se izražanje genov za mlečne proteine po imunostimulaciji večinoma ni spremenilo, le pri PAEP se je zmanjšalo, pri CSN1S1 pa celo povečalo. Naše ugotovitve lahko pripomorejo k boljšemu razumevanju imunskega odziva mlečne žleze, predvsem začetnih faz aktivacije vnetnega odziva preko epitelijskih celic, ter lahko predstavljajo izhodišče za nadaljnje študije mastitisa in vitro. Mastitis is an inflammatory disease of the mammary gland, which affects almost half of all dairy cattle and thus leads to huge financial losses in the dairy industry. The disease can lower milk yield and milk quality significantly, as it can affect the amount and destabilize the ratio between milk proteins. Various pathogens can cause mastitis, among which E. coli and S. aureus are the most common. In this work, the effect of lipopolysaccharide (LPS) and lipoteichoic acid (LTA), the key components of pathogens’ cell wall and virulent factors, on immune response and protein synthesis in mammary gland was comparatively investigated. For this purpose, primary bovine mammary epithelial cells were successfully cultured in vitro and stimulated for 24 hours with LPS and LTA, respectively. The qPCR results showed the gene expression of inflammatory factors was increased after overnight stimulation, where LPS led to greater increase than LTA. ELISA demonstrated higher concentrations of IL-8 and TNF-α in LPS-stimulated cultures, whereas LTA decreased them. Established cultures were able to synthesize milk proteins, as it was shown with immunostaining. Differential gene expression analysis showed no changes in gene expression for most milk proteins, only PAEP was downregulated, while CSN1S1 was upregulated. Our findings could lead to better understanding of immune response in mammary gland, especially the early stages of activation of inflammatory response through epithelial cells, and could represent the basis for subsequent studies of mastitis in vitro. goveja mlečna žleza izražanje genov mastitis mlečni proteini prirojena imunost bovine mammary gland gene expression innate immunity mastitis milk proteins true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-08-26 10:40:06 2026-08-26 10:40:15 2026-09-09 08:58:00 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25630 290446851 Puhov_Spela_-_Vpliv_imunostimulacije_primarne_epitelijske_celicne_kulture_mlecne_zleze_goveda_z_.pdf Puhov_Spela_-_Vpliv_imunostimulacije_primarne_epitelijske_celicne_kulture_mlecne_zleze_goveda_z_.pdf 1 5F471094DE4BDB811368E2A9B7F0680B bd90ad1df10519a8a5f10a396c916537a597b86a4b4c93b8b24513ed269a874d 5940feb5-a129-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245111 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0