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Title:Vpliv onesnažil iz okoljskih aerosolov na tarčne celice v pljučih
Authors:Pribovšek, Sara (Author)
Bergant Marušič, Martina (Mentor) More about this mentor... New window
Files:.pdf RAZ_Pribovsek_Sara_i2016.pdf (7,73 MB)
 
Language:Slovenian
Work type:Bachelor thesis/paper (mb11)
Tipology:2.11 - Undergraduate Thesis
Organization:FZO - Faculty of Environmental Sciences
Abstract:Diplomsko delo obravnava vpliv onesnažil iz okoljskih aerosolov na dva celična tipa, ki jih najdemo v pljučih, pljučne epitelijske celice A549 in makrofage J774A.1. Kot vir onesnažil smo uporabili ekstrakt saj iz zračnega filtra. Uporabili smo kvarčne zračne filtre z delci črnega ogljika, ki so nastali kot posledica gorenja sveče. V laboratorijskih poskusih smo testirali neposredni vpliv saj na celično živost (viabilnost) pljučnih epitelijskih celic in makrofagov po 24 urah in 48 urah inkubacije z različnimi koncentracijami saj in kontrolama. Celično živost smo merili s pomočjo reagenta Presto Blue. Posreden vpliv saj na pljučne epitelijske celice smo preverjali z merjenjem celične živosti po 24 urah in 48 urah inkubacije s topnimi dejavniki, ki so jih sprostili tretirani in kontrolni makrofagi (gojišče MCM). Gojišče MCM smo pripravili tako, da smo makrofage 48 ur aktivirali z raztopino saj oziroma jih nismo aktivirali (kontrolno gojišče MCM). Na koncu smo pri obeh celičnih tipih ugotavljali tudi gensko stabilnost s pomočjo posredne imunofluorescentne metode. Celice smo označili s protitelesi anti-fosfo histon H2A.X, ki so se vezala na mesta, kjer je zaradi izpostavljenosti raztopini saj prišlo do poškodb DNA. Rezultati so pokazali, da raztopina saj vpliva na celično živost pljučnih epitelijskih celic in makrofagov v obratno sorazmernem trendu v odvisnosti od koncentracije saj. Raztopina saj je imela vpliv tudi na morfološke spremembe obeh celičnih linij. Rezultati vpliva topnih dejavnikov makrofagov na epitelijske pljučne celice niso pokazali statistično značilnih razlik, zato posrednega vpliva saj na viabilnost pljučnih celic zaradi kompleksnosti delovanja topnih faktorjev makrofagov nismo dokazali. Pokazali pa smo, da imajo saje velik vpliv tudi na gensko stabilnost pljučnih epitelijskih celic in makrofagov. V diplomskem delu smo tako pokazali, da že krajša izpostavljenost onesnažilom iz zraka vodi do poškodb DNA in zmanjšane celične živosti tarčnih celic, kar je bilo še zlasti izrazito pri makrofagih.
Keywords:makrofagi, pljučne epitelijske celice, delci, policiklični aromatski ogljikovodiki, celična živost, genotoksičnost.
Year of publishing:2016
Source:Nova Gorica
COBISS_ID:4159739 Link is opened in a new window
URN:URN:SI:UNG:REP:1ZF2INUS
Views:3558
Downloads:109
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Secondary language

Language:Undetermined
Title:Impact of contaminants from environmental aerosols on the target cells in the lungs
Abstract:This work, entitled as »Impact of contaminants from environmental aerosols on the target cells in the lungs«, addresses the effects of environmental pollutants of aerosols on two cell types that are found in the lungs, pulmonary epithelial cells A549 and macrophages J774A.1. We used an air filter extract as a source of pollutants. Quartz filters containing black carbon particles from burning a candle were used in experiments. In laboratory experiments, we have tested direct effects of soot on cell viability of lung epithelial cells and macrophages after 24 h and 48 h of incubation with different concentrations of soot and controls. Cell viability was measured using a Presto Blue reagent. The indirect soot effect on lung epithelial cells were examined by measuring cell viability after 24 h and 48 h incubation with soluble factors released by treated and control macrophages (MCM medium). The MCM medium was prepared via activation of macrophages which were exposed to a solution of soot for 48 h and to growth medium for control MCM medium. In the end, we determined genetic stability of both cell types using the indirect immunofluorescence method. Cells were tagged with anti-phospho histone H2A.X antibodies, that bound to positions where DNA damage caused due to exposure to soot solution. The results have shown that the solution of soot affected the cell viability of lung epithelial cells and macrophages, in a dose-dependent manner. Soot solution also had an influence on morphological changes in both cell lines. The results of the impact of soluble factors of macrophages in lung epithelial cells did not show statistically significant differences, therefore the indirect influences of soot on viability of lung cells were not confirmed due to the complex acting nature of soluble factors released from macrophages. However we have shown that the environmental pollutants of aerosols had a large influence on the genetic stability of lung epithelial cells and macrophages. In this work we have also shown that even short exposure to air pollutants leads to DNA damage and lower cell viability of the target cells, which was especially notable in the macrophages.
Keywords:macrophages, lung epithelial cells, particulate matter, polycyclic aromatic hydrocarbons, cellular viability, genotoxicity.


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