20.500.12556/RUL-174230 Vpliv spola donorja limfoblastoidne celične linije na imunomodulatorne učinke izbranih bisfenolov Influence of the gender of the lymphoblastoid cell line donor on the immunomodulatory effects of selected bisphenols Bisfenol A (BPA) in njegovi analogi so kemikalije, ki se uporabljajo pri proizvodnji plastike in epoksidnih smol, vendar je zaradi toksičnih učinkov njihova uporaba strogo nadzorovana. Delujejo kot motilci endokrinega sistema, saj vplivajo na delovanje hormonov, zlasti spolnih, preko katerih povzročajo različne biološke učinke, vključno z imunomodulatornimi. Zaradi njihovega vpliva na spolne hormone nas je zanimalo, ali obstajajo razlike v imunskem odzivu med spoloma po izpostavitvi izbranim bisfenolom. Za preučevanje smo izbrali sedem bisfenolov, in sicer BPA, BPE, BPAF, BPAP, BPPH, TMBPF ter TCBPA. Kot model in vitro smo uporabili limfoblastoidne celične linije (LCL) zdravih, nesorodnih donorjev, in sicer pet LCL ženskih darovalk ter pet LCL moških darovalcev. Najprej smo določili citotoksičnost izbranih spojin s testom presnovne aktivnosti MTS in izračunali srednje inhibitorne koncentracije - vrednosti IC50. Na podlagi povprečnih vrednosti IC50, določenih v desetih celičnih linijah LCL, so bile za preučevane bisfenole ugotovljene naslednje vrednosti (od najmanj do najbolj citotoksičnega): BPE (IC50 = 156,8 μM), TCBPA (IC50 = 124,7 μM), BPA (IC50 = 89,3 μM), BPAP (IC50 = 59,0 μM), BPAF (IC50 = 31,8 μM), BPPH (IC50 = 31,8 μM) in TMBPF (IC50 = 9,2 μM). S primerjavo vrednosti IC50 in logP smo ugotovili, da ti dve vrednosti med sabo korelirata, in sicer so bolj lipofilne spojine tudi bolj citotoksične. Naredili smo korelacijsko analizo med posameznimi vrednostmi IC50 bisfenolov v desetih testiranih LCL in ugotovili, da je najvišja korelacija med BPAF ter TCBPA. Ugotovili smo, da so pri vseh preučevanih bisfenolih povprečne vrednosti IC50 višje v celicah LCL moških v primerjavi s celicami LCL ženskih darovalk. Pri preučevanju imunomodulatornih lastnosti izbranih bisfenolov z merjenjem izločenih citokinov smo ugotovili, da je vpliv na modulacijo izločanja citokinov odvisen od koncentracije spojine. Po izpostavitvi 10 μM koncentracijam so imele največji vpliv na izločanje citokinov spojine BPA, BPAF, TMBPF in BPPH, ki je izmed vseh deloval najbolj imunosupresivno. Bisfenoli v 100 nM koncentraciji so imeli nižji vpliv na modulacijo izločanja citokinov. Pri analizi interindividualnih razlik je bil opaženi spolno specifičen trend imunosupresivnega delovanja bisfenolov, z izrazitejšim učinkom pri moških, vendar brez statistično značilne razlike. Iz rezultatov sklepamo, da bisfenoli izkazujejo različno stopnjo citotoksičnosti in imunomodulatornega delovanja, pri čemer so učinki odvisni tako od strukture spojine kot tudi od koncentracije. Opazili smo tudi spolno specifične trende v odzivu celic LCL, kar nakazuje na potencialno vlogo spolnih hormonov pri modulaciji učinkov teh spojin. Bisphenol A (BPA) and its analogues are chemicals used in the production of plastics and epoxy resins, but due to their toxic effects, their use is strictly regulated. They act as endocrine-disrupting chemicals, as they interfere with hormone function – particularly sex hormones – through which they cause various biological effects, including immunomodulatory ones. Because on their impact on sex hormones, we investigated whether there are differences in immune responses between genders following exposure to selected bisphenols. We selected seven bisphenols for investigation: BPA, BPE, BPAF, BPAP, BPPH, TMBPF, and TCBPA. As an in vitro model, we used lymphoblastoid cell lines (LCLs) from healthy, unrelated donors – specifically, five female and five male LCLs. First, we determined cytotoxicity of the selected compounds using the MTS assay and calculated the IC50 values. Based on the average IC50 values determined in ten LCL cell lines, the following IC50 values were obtained for the bispenols studied (from least to most cytotoxic): BPE (IC50 = 156,8 μM), TCBPA (IC50 = 124,7 μM), BPA (IC50 = 89,3 μM), BPAP (IC50 = 59,0 μM), BPAF (IC50 = 31,8 μM), BPPH (IC50 = 31,8 μM) and TMBPF (IC50 = 9,2 μM). Correlation analysis between IC₅₀ values and log P indicated that increased lipophilicity was associated with enhanced cytotoxic activity. A correlation analysis of IC50 values of bisphenols across ten tested LCLs revealed the highest correlation between BPAF and TCBPA. The average IC50 values were higher in LCLs derived from male donors compared to those from female donors. Evaluation of the immunomodulatory properties of the selected bisphenols using cytokine release assays revealed that their effects were dose-dependent. The most pronounced immunomodulatory effects were observed for BPA, BPAF, TMBPF, and BPPH at higher, 10 μM concentration. The bisphenols caused a minor impact on cytokine secretion at 100 nM concentration. We observed sex-specific immunomodulatory effects of tested bisfenols, indicating that bisphenols may act more immunosuppressive in male cells. In conclusion, bisphenols exhibited varying degrees of cytotoxicity and immunomodulatory activity, with effects depending on both the chemical structure of the compound and its concentration. We observed sex-specific trends in the response to bisphenols in LCL cells, indicating a potential role of sex in bisphenol-induced immunotoxicity. bisfenol citotoksičnost imunomodulacija interindividualne razlike limfoblastoidne celične linije bisphenol cytotoxicity immunomodulation interindividual differences lymphoblastoid cell lines true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-09-30 08:46:19 2025-09-30 08:46:24 2025-11-19 03:50:12 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 2026-09-29 119523 30112.pdf 30112.pdf 1 B5699EC31C8A96AC5640BE11ACD373C0 f002fce3260255fa9c35ccacfabe8905a94bcc3dc7d32182e5272a43942606fb fac93a33-9dc8-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=218982 Fakulteta za farmacijo 0 0 0