Ispitivanje citotoskičnosti ekstrakata micelija plijesni vrsta Aspergillus piperis i Aspergillus luchuensis u stanicama ljudskog adenokarcinoma pluća A549

master's thesis

• Title (english): Cytotoxicity of mold mycelium extracts of Aspergillus piperis and Aspergillus luchuensis species on A549 human lung adenocarcinoma cells
• Author: Ida Ruszkowski
• Mentor: Daniela Jakšić (mentor)
• Committee member: Daniela Jakšić (predsjednik povjerenstva)
• Committee member: Ana-Marija Domijan (član povjerenstva)
• Committee member: Maja Friščić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of microbiology); Zagreb
• Defense date and country: 2020-06-05, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Crni aspergili široko su rasprostranjene plijesni prilagođene različitim supstratima. Pored primjene u biotehnologiji i industriji, mogu biti i prijetnja zdravlju s obzirom na alergijski, invazivni i toksični potencijal. Cilj ovog rada bio je usporediti toksičnost ekstrakata micelija različitih izolata vrsta Aspergillus piperis i Aspergillus luchuensis s obzirom na morfološku i filogenetičku srodnost te zbog pojavnosti spomenutih vrsta u zraku i prašini zatvorenih prostora preko kojih mogu u dišnom sustavu izloženih ljudi djelovati štetno. U ovome radu ispitan je citotoksični učinak ekstrakata micelija 9 izolata vrsta A. piperis i A. luchuensis u stanicama ljudskoga adenokarcinoma pluća A549. Ispitivanje se provodilo tretiranjem A549 stanica različitim koncentracijama ekstrakata micelija (0,025–0,8 mg/ml) svakog izolata tijekom 24 sata. Vijabilnost tretiranih stanica određena je MTT testom citotoksičnosti u odnosu na kontrole te su izračunate koncentracije koje inhibiraju rast 50% stanica - IC50. Svi ispitani ekstrakti micelija pokazali su se citotoksičnima u A549 stanicama među kojima se najtoksičniji pokazao onaj priređen od izolata vrste A. piperis MFBF AN12614 koji je pri najmanjoj ispitanoj koncentraciji 0,025 mg/ml za 90% smanjio vijabilnost A549 stanica. Najmanje toksičan pokazao se ekstrakt micelija A. piperis MFBF AN12404 s IC50 od 0,5068 mg/ml. Ekstrakti micelija vrste A. luchuensis pokazali su izraženi toksični potencijal te su IC50 vrijednosti sva četiri ispitana ekstrakta bile niže od 0,1 mg/ml, dok je s obzirom na izračunatu IC50 od 0,03 mg/ml ekstrakt izolata MFBF AN10927A iz zraka iz mlina žitarica bio najtoksičniji među ispitanim ekstraktima micelija vrste A. luchuensis. Rezultati ovoga rada upućuju na kompleksnost sastava metabolita u ekstraktima pojedinih izolata vrsta A. piperis i A. luchuensis obzirom da je toksični potencijal obilježje pojednoga izolata neovisno o vrsti. Time su dobiveni rezultati usklađeni s visokim stupnjem filogenetičke srodnosti. Budući da su se svi izolati spomenutih vrsta pokazali citotoksičnima u A549 stanicama neovisno o izvoru izolata, postoji opasnost od toksičnoga učinka pri inhalaciji toksinogenih mikočestica micelija u prostorima gdje ljudi borave.

Abstract (english)

Black Aspergilli are widespread molds inhabiting different substrates. In addition to their many positive roles in biotechnology and food industry, they can also be harmful because of their allergic, invasive and toxic potential. This paper aimed to compare the cytotoxicity of mycelium extracts of different isolates of A. piperis and A. luchuensis species because of their morphological and phylogenetic relatedness and because of their incidence in the air and dust in indoor environments in which they can be harmful to the respiratory system of exposed people. In this paper was tested the cytotoxicity of mycelium extracts of nine isolates of A. piperis and A. luchuensis species on A549 human lung adenocarcinoma cells. The assay was performed by treating A549 cells with different concentrations (0.025 - 0.8 mg/ml) of mycelium extracts of each isolate for 24 hours. The viability of treated cells was measured by MTT cytotoxicity assay relative to the controls, and the concentrations inhibiting the growth of 50% cells - IC50 were calculated. All tested mycelium extracts proved to be cytotoxic on A549 cells. The most toxic was the mycelium extract of A. piperis isolate MFBF AN12614 which decreased viability of A549 cells by 90% at a concentration of 0.025 mg/ml. The least toxic was mycelium extract of A. piperis MFBF AN12404 with IC50 of 0.5068 mg/ml. Mycelium extracts of A. luchuensis species showed pronounced toxic potential and IC50 of all four tested mycelium extract were lower than 0.1 mg/ml. Considering IC50 of 0.03 mg/ml, extract of airborne isolate MFBF AN10927A from a grain mill was the most toxic among the examined mycelium extracts of A. luchuensis species. The results of this study indicate the complexity of metabolite composition in extracts of individual isolates of A. piperis and A. luchuensis species and the toxic potential is characteristic of an individual isolate regardless of species. Thereby, the results are consistent with a high degree of phylogenetic relatedness. Because all tested isolates of A. piperis and A. luchuensis proved to be cytotoxic on A549 cells, inhalation of airborne or dustborne toxinogenic mycoparticles may cause damage in respiratory system of exposed people, regardless of the type of indoor environment or water damage.

• Keywords: Aspergillus piperis; Aspergillus luchuensis; citotoksičnost; ekstrakt micelija; A549 stanice; IC50
• Keywords (english): Aspergillus piperis; Aspergillus luchuensis; cytotoxicity; mycelium extracts; A549 cells; IC50
• Language: croatian
• URN:NBN: urn:nbn:hr:163:438172
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-11-15 16:35:30