Preučevanje imunomodulatornega delovanja zmesi bisfenola A in njegovih nadomestkov v limfoblastoidnih celičnih linijah

Knap, Larisa (author), Sollner Dolenc, Marija (mentor), Markovič, Tijana (comentor)

Bisfenol A je snov, ki se največ uporablja v proizvodnji polikarbonatne plastike in epoksi smol. V raziskavah so že bili ugotovljeni njegovi endokrini, kancerogeni, mutageni in imunotoksični učinki. Vpliva pa tudi na povečanje tveganja za razvoj nekaterih bolezni. Zato se je pojavila potreba po nadomestkih bisfenola A, ki bi imeli enake funkcionalne lastnosti, vendar z manj neželenimi učinki na žive organizme. Zanimalo nas je njihovo citotoksično in imunomodulatorno delovanje v primerjavi z bisfenolom A ter kakšne so imunomodulatorne lastnosti zmesi bisfenola A z nadomestki. Preučevali smo 16 nadomestkov bisfenola A: bisfenol AF, bisfenol AP, bisfenol B, bisfenol C, bisfenol E, bisfenol F, bisfenol G, bisfenol P, bisfenol PH, bisfenol S-monoalil eter, bisfenol S-monobenzil eter, bisfenol Z, BTUM, Pergafast 201, tetrametil bisfenol F in 2,4-bisfenol S. Učinke spojin na imunske celice smo vrednotili na limfoblastoidnih celičnih linijah treh različnih donorjev, ki predstavljajo in vitro model za vrednotenje imunomodulatornih lastnosti spojin. Najprej smo preučevali citotoksičnost za limfoblastoidne celične linije nadomestkov v primerjavi z bisfenolom A in ugotovili, da je bilo 6 nadomestkov bisfenola A manj, 10 pa bolj citotoksičnih od bisfenola A. Najbolj citotoksične spojine so imele srednjo inhibitorno koncentracijo 19,5 µM (bisfenol G), 28,6 µM (bisfenol P) in 30,6 µM (bisfenol PH). Nadalje smo ugotovili, da srednja inhibitorna koncentracija spojin korelira z logaritmom porazdelitvenega koeficienta spojin, pri čemer je večja citotoksičnost bolj lipofilnih spojin predvidoma povezana z boljšim in lažjim prehajanjem spojin preko membran celic. Preučevali smo tudi vpliv posameznih nadomestkov in binarnih zmesi bisfenola A s 5 nadomestki na modulacijo izločanja citokinov. Pri 10 µM koncentraciji so binarne zmesi imele večji vpliv na izločanje citokinov kot posamezni bisfenoli. Najbolj je izstopala zmes bisfenola A/bisfenola AP, ki je močno znižala izločanje interlevkinov 2, 6, 10 in dejavnika tumorske nekroze alfa. V zadnjem delu smo na celični liniji Ramos-Blue potrdili nemonotono delovanje bisfenolov. Ti so pri 10 µM koncentraciji znižali aktivnost signalne poti jedrnega dejavnika kapa B/proteinskega aktivatorja 1, pri 100 nM koncentraciji pa zvišali.

• Language: Slovenian
• Keywords: bisfenol A, nadomestki bisfenola A, limfoblastoidna celična linija, Ramos-Blue celična linija, citokini
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-163028
• Publication date in RUL: 01.10.2024

Secondary language

• Language: English
• Title: Immunomodulatory activity evaluation of mixtures of bisphenol A and its substitutes in human lymphoblastoid cell lines

Abstract

Bisphenol A is most commonly used in the production of polycarbonate plastic and epoxy resin. Its endocrine, carcinogenic, mutagenic effects and immunotoxic effects have already been established. It also increases the risk of developing certain diseases. Due to these facts, there was a need for bisphenol A substitutes, which would have similar functionality but fewer adverse effects on living organisms. We were interested in comparing their cytotoxic and immunomodulatory effects to bisphenol A, as well as the immunomodulatory properties of mixtures of bisphenol A with its substitutes. We studied 16 bisphenol A substitutes: bisphenol AF, bisphenol AP, bisphenol B, bisphenol C, bisphenol E, bisphenol F, bisphenol G, bisphenol P, bisphenol PH, bisphenol S-monoalyl ether, bisphenol S-monobenzyl ether, bisphenol Z, BTUM, pergafast 201, tetramethyl bisphenol F and 2,4-bisphenol S. The effects of these compounds on immune cells were evaluated on lymphoblastoid cell lines from three unrelated donors. These cell lines represent in vitro model for assessing the immunomodulatory properties of compounds. First, we studied the cytotoxicity of the substitutes and found that six substitutes were less, while ten were more cytotoxic than bisphenol A. The most cytotoxic compounds had the lowest inhibitory concentration, 19,5 µM (bisphenol G), 28,6 µM (bisphenol P), and 30,6 µM (bisphenol PH). We determined that half-maximal inhibitory concentration correlates with logarithm of the partition coefficient, suggesting that higher cytotoxicity of more lipophilic compounds is likely related to better and easier membrane permeability. We also studied effects of individual bisphenol A substitutes and binary mixtures of bisphenol A with five substitutes on the modulation of cytokine release. Binary mixtures had stronger impact on modulation of cytokine release than individual bisphenols at 10 µM concentration. The bisphenol A/bisphenol AP mixture stood out the most, significantly inhibiting the secretion of interleukine 2, 6, 10, and tumor necrosis factor alpha. In the last part, we found a non-monotonic effect of bisphenols on the Ramos-Blue cell line, 10 µM concentration decreased the activity of nuclear factor κB/activator protein 1, while 100 nM concentration increased it.

• Keywords: bisphenol A, bisphenol A substitutes, lymphoblastoid cell lines, Ramos-Blue cell line, cytokines