Vpliv izbranih nanomaterialov na aktivnost acetilholinesteraze AChE v celični kulturi

Javornik, Tjaša (avtor), Drobne, Damjana (mentor)

Ob stiku nanomaterialov (NM) z biološkim medijem, površina delca adsorbira molekule iz okolja. Te molekule oblikujejo korono, ki določa lastnosti in vpliv NM v bioloških sistemih. Namen magistrske naloge je bil preveriti vpliv izbranih NM s proteinsko korono na aktivnost encima acetilholinesteraza (AChE) v celicah in vitro. Za testiranje aktivnosti encima smo uporabili Ellmanovo metodo. Celice SH-SY5Y smo za 24 ur izpostavili nanodelcem (ND) TiO2, SiO2, ZnO in TiOx. S spektrofotometrom smo nato pri valovni dolžini 405 nm izmerili količino rumenega produkta, ki nastane ob reakciji AChE z acetiltioholinkloridom. Vpliv ND TiO2, ZnO in TiOx s predhodno oblikovano korono je odvisen od koncentracije, medtem ko je pri ND SiO2 s predhodno oblikovano korono vpliv pri vseh koncentracijah podoben. Vpliv TiO2 (oznake E171-E, G10-4, 67-CC) in ZnO ND brez predhodno oblikovane korone je odvisen od koncentracije. Ostali ND brez predhodno oblikovane korone imajo pri vseh koncentracijah podoben vpliv. Vpliv ND na delovanje encima AChE je odvisen od tipa korone, ki je bila pri predhodno oplaščenih ND iz govejega serumskega albumina. ND brez predhodne korone so se obdali z molekulami ob stiku z medijem DMEM. Večina ND s predhodno korono je povečala aktivnost AChE, medtem ko se je pri tretiranju z ND brez predhodne korone aktivnosti encima pri večini ND zmanjšala. Za ND brez predhodno oblikovane korone sklepamo, da je bil privzem z endocitozo zmanjšan zaradi tvorbe agregatov, vendar tega nismo preverili.

• Jezik: Slovenski jezik
• Ključne besede: nanomateriali, AChE, korona, TiO2, SiO2, ZnO, TiOx, SH-SY5Y
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: BF - Biotehniška fakulteta
• Leto izida: 2020
• PID: 20.500.12556/RUL-120436
• COBISS.SI-ID: 33544451
• Datum objave v RUL: 20.09.2020

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Effect of selected nanomaterials on the activity of acetylcholinesterase (AChE) in cell culture

Izvleček

When nanomaterials (NM) come in contact with the biological medium, the particle surface adsorbs molecules from the environment. These molecules form a corona that determines the characteristics and impact of NM on biological systems. The purpose of this master's thesis is to study the impact of selected NM with a protein corona on the activity of the acetylcholinesterase (AChE) enzyme in cells in vitro. Ellman's method was used to test the enzyme activity. SH-SY5Y cells were exposed to TiO2, SiO2, ZnO, and TiOx nanoparticles (NP) for 24 hours. Yellow product formed by the reaction of AChE with acetylthiocholine chloride was measured with a spectrophotometer at 405 nm. The impact of TiO2, ZnO, and TiOx NP with formed corona is dose-dependent, while in NP SiO2 with formed corona the impact on the enzyme is similar at all concentrations. The impact of TiO2 (E171-E, G10-4, 67-CC) and ZnO NP without formed corona is also dose-dependent. The other NP without formed corona had a similar impact on the enzyme at all concentrations. The impact of NP on the activity of the AChE enzyme depends on the type of corona. NP with formed corona were coated with bovine serum albumin. The NP without prior corona were surrounded by molecules upon contact with the DMEM medium. Most NP with bovine serum albumin corona increased AChE activity, whereas NP without prior corona decreased the enzyme activity. For NP without prior corona, we suggest that uptake by endocytosis was reduced due to aggregate formation, but this hypothesis was not verified.

• Ključne besede: nanomaterials, AChE, corona, TiO2, SiO2, ZnO, TiOx, SH-SY5Y