Vpliv atmosferskih aerosolov na fiziološki in vnetni odziv makrofagov

magistrsko delo

Tina Čujec (Author), Martina Bergant Marušič (Mentor)

Abstract

Magistrsko delo obravnava vpliv aerosoliziranih delcev na fiziološki in vnetni odziv makrofagov J774.2. Makrofage smo v prvem delu izpostavili suspenziji filtra z delci črnega ogljika, v drugem delu pa smo postopek izpostavitve ponovili s suspenzijo filtrov, na katere so bili vzorčeni delci iz zunanjega zraka. V obeh primerih smo celice izpostavili različnim koncentracijam suspenzij in analizirali njihov vpliv po 24- urni ali 48- urni inkubaciji. Celično viabilnost smo določili z reagentom PrestoBlue, gensko stabilnost smo ugotavljali z imunofluorescenčno metodo (anti-fosfo-histon H2A.X), oksidativni stres pa smo kvantificirali z barvilom karboksi-H2DCFDA. Vnetni odziv izpostavljenih celic smo opazovali s testom fagocitoze in določanjem citokina TNFα (test ELISA). Rezultati so pokazali, da suspenzija črnega ogljika vpliva na celično živost makrofagov v odvisnosti od koncentracije dodane suspenzije. Dokazali smo povečan nastanek prostih kisikovih radikalov (ROS), znižano gensko stabilnost, ter povečan imunski odziv izpostavljenih celic. Delci črnega ogljika so bistveno povečali obseg fagocitoze in izločanje vnetnega citokina TNFα pri izpostavljenih makrofagih. Pri vseh testih se je odziv večal z višjimi koncentracijami suspenzije črnega ogljika. Rezultati analiz s suspenzijo realnih vzorcev so pokazali, da je vnetni imunski odziv eden prvih odgovorov makrofagov na izpostavljenost okoljskim aerosolom. Opazili smo tudi večje število poškodb DNA izpostavljenih makrofagov, medtem ko je bila viabilnost teh celic še neprizadeta. Na osnovi rezultatov magistrske naloge smo zaključili, da okoljski aerosoli sprožijo vnetni odziv izpostavljenih makrofagov, negativno pa vplivajo tudi na njihovo gensko stabilnost, viabilnost in nastanek ROS. Predvidevamo, da je izraženost učinkov odvisna tudi od koncentracije in sestave okoljskih aerosolov.

Keywords

makrofagi;črni ogljik;okoljski aerosoli;vnetni odziv;genotoksičnost;magistrske naloge;

Data

Language:	Slovenian
Year of publishing:	2017
Typology:	2.09 - Master's Thesis
Organization:	UNG FZO - Faculty of Environmental Sciences
Publisher:	[T. Čujec]
UDC:	54
COBISS:	4930043
Views:	4876
Downloads:	280
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	The influence of atmospheric aerosols on physiological and inflammatory responses of macrophages.
Secondary abstract:	The master thesis analyzes the influence of atmospheric aerosols on physiological and inflammatory responses of macrophages J774.2. In first part, the macrophages were exposed to the black carbon suspension. In second part, the exposure procedure was repeated with a suspension of atmospheric aerosols. In both cases, cells were exposed to various concentrations of suspensions. The effects were analyzed after a 24- hour or 48- hour incubation. Cell viability was determined by the PrestoBlue reagent, genetic stability was determined by the immunofluorescence method (anti-phospho-histon H2A.X) and oxidative stress was quantified with carboxy-H2DCFDA dye. Inflammatory responses of exposed cells were evaluated with the phagocytosis assay and with secretion of cytokine TNFα (ELISA assay). We confirmed that cell viability of macrophages decreases with the increasing concentrations of the black carbon suspension. We also observed increased ROS formation, reduced genetic stability and increased immune response after the exposure. Black carbon particles have significantly increased the extent of phagocytosis and the secretion of inflammatory cytokine TNFα in exposed macrophages. Experiments with the suspension of atmospheric aerosols have shown that inflammatory immune response is one of the the primary responses of macrophages exposed to aerosols. The phagocytosis and ELISA assays showed concentration - dependent inflammatory response. We also observed DNA damage of the exposed macrophages, while the viability was not yet affected. We concluded that atmospheric aerosols activate immune system response of the exposed macrophages and also have negative effect on genetic stability, viability and ROS formation. We assume that effect extent of macrophages coused by atmospheric aerosols is concentration and composition dependent.
Secondary keywords:	Macrophages;black carbon;atmospheric aerosols;inflammatory response;genotoxicity.;
URN:	URN:SI:UNG
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Novi Gorici, Fak. za znanosti o okolju
Pages:	XI, 52 str.
ID:	10869016