Določanje učinka avobenzona, butiliranega hidroksianizola in 2-metilresorcinola na androgenih in glukokortikoidnih receptorjih
| Autor: | Madžarac, Vanja |
|---|---|
| Přispěvatelé: | Sollner Dolenc, Marija |
| Jazyk: | slovinština |
| Rok vydání: | 2020 |
| Předmět: |
hormonski motilci hormonski sistem androgeni receptor glukokotrikoidni receptor avobenzon 2-metilresorcinol butiliran hidroskianizol MTS test luciferazni test celična linija MDA-kb2
endocrine disruptors endocrine system androgen receptor glucocorticoid receptor avobenzone 2-methylresorcinol butylated hydroxyanisole cell line MDA-kb2 MTS test luciferase assay udc:577.175+615.357(043.2) |
| Popis: | Hormonski motilci so eksogene snovi, ki vplivajo na delovanje hormonskega sistema najpogosteje z vezavo na endokrine receptorje in lahko njegov odziv povečajo ali ga zavrejo. Do sedaj so z raziskavami dokazali endokrine učinke pri velikem številu spojin, katerim smo izpostavljeni vsakodnevno, nekatere so tudi v kozmetičnih izdelkih. Raziskave kažejo, da kljub uporabljenim nizkim koncentracijam kozmetičnih sestavin z endokrinim delovanjem v izdelkih njihovi učinki niso zanemarljivi, saj vezava liganda z receptorjem povzroči kaskado reakcij v celici, kjer je vsaka naslednja stopnja prenosa informacije močnejša. V diplomski nalogi smo se odločili preveriti endokrini učinek treh pogosto uporabljenih spojin v kozmetičnih izdelkih. Testirali smo UV filter avobenzon (AVB), prekurzor oksidativnih barv za lase 2-metilresorcinol (2MR) in antioksidant butiliran hidroksianizol (BHA). Področje njihovega delovanja na endokrini sistem je še dokaj neraziskano, predvsem vpliv na moduliranje delovanja androgenega (AR) in glukokortikoidnega (GR) receptorja, na kar smo se osredotočili v svoji raziskavi. In vitro test smo izvajali na celični liniji MDA-kb2, ki ima izražen AR in GR hkrati ter omogoča razlikovanje med agonističnim in antagonističnim učinkom spojin. Najprej smo s citotoksičnim MTS testom določili prvo necitotoksično koncentracijo izbrane spojine in tako določili primerne koncentracije za nadaljnji luciferazni test, ki vrednoti učinke spojin na določenem receptorju preko ekspresije luciferaze, ki jo lahko testirane spojine povečajo ali zmanjšajo. V raziskavi smo zabeležili statistično značilne učinke spojin, in sicer na AR smo zabeležili antagonistično delovanje AVB pri 0,01 μM in 0,0001 μM, 2MR pri 175 μM ter BHA pri 50 μM, 0,5 μM in 0,01 μM koncentraciji, medtem ko nobena od spojin ni delovala AR agonistično. Vse spojine so tudi izkazale agonističen učinek na GR, in sicer AVB pri 1 μM, 0,01 μM in 0,0001 μM, 2MR pri 175 μM, 1,75 μM in 0,01 μM ter BHA pri 50 μM, 0,5 μM in 0,01 μM koncentraciji. Antagonizem na GR sta izkazala samo 2MR pri 175 μM in BHA pri 50 μM koncentraciji. V naši raziskavi so vse testirane spojine izkazale učinke na AR in GR. Za potrditev naše domneve o vplivu AVB, 2MR in BHA na delovanje hormonskega sistema so potrebne še nadaljnje podrobnejše raziskave. Na osnovi pridobljenih rezultatov v tej raziskavi pa svetujemo previdnost pri njihovi uporabi tudi zaradi možnega vpliva na endokrini sistem. Endocrine disrupters are exogenous chemicals which interfere with endocrine system by binding to certain hormone receptor and consequently enhance or reduce receptor’s activity. Several studies have so far proven, for a number of chemicals, which are used daily, to have endocrine effects. Among them, several are used in cosmetics. Although low concentrations of chemicals with endocrine effect are used in cosmetics, there are studies, showing that effects of low concentrations are not negligible, as binding of ligand with receptor, causes a cascade of reactions, where every next stage of transferring information, is stronger. In our study we have decided to focus on endocrine effects of three frequently used compounds in cosmetics. We have tested UV filter avobenzone (AVB), precursor of oxidative hair dyes 2-methylresorcinol (2MR) and antioxidant butylated hydroxyanisole (BHA). As endocrine disrupting effect of these compounds is not well researched, especially on androgen (AR) and glucocorticoide (GR) receptor, we have decided to investigate this area. Our in vitro test was performed on MDA-kb2 cell line, where both AR and GR are presented at the same time, with the ability to differentiate between agonist and antagonist effect of the compounds. At first we defined the first non-cytotoxic concentration of the compounds and determined suitable concentrations for further analyses, with the MTS test. Afterwards we started with the luciferase assay, where the effect of the compound on specific receptor, is measured through expression of luciferase, where compounds can increase or reduce the luciferase activity. In our research we have noted significant effects of our tested compounds, especially on AR we have noted an antagonistic effect of AVB at 0,01 μM and 0,0001 μM, antagonistic effect of 2MR at 175 μM and antagonistic effect of BHA at 50 μM, 0,5 μM and 0,01 μM concentration, while none of the compounds showed agonist effect on AR. All of the compounds as well showed agonist effect on GR, specifically AVB at 1 μM, 0,01 μM and 0,0001 μM, 2MR at 175 μM, 1,75 μM and 0,01 μM and BHA at 50 μM, 0,5 μM and 0,01 μM concentration. GR antagonistic effect was only noted for 2MR at 175 μM and BHA at 50 μM. Our research has indicated effects on AR and GR by all of the tested compounds. For final confirmation of our presumption of AVB, 2MR and BHA effect on endocrine system, we need more thorough analysis. Based on our research, we suggest caution when using products containing these compounds, as they may interfere with the endocrine system. |
| Databáze: | OpenAIRE |
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