20.500.12556/RUL-175186 Protivnetni in protifibrotični učinki kordicepina in ekstraktov gliv Cordyceps militaris ter Hirsutella sinensis na monocite in fibroblaste Anti-inflammatory and anti-fibrotic effects of cordycepin and extracts from the fungi Cordyceps militaris and Hirsutella sinensis on monocytes and fibroblasts Glivi Cordyceps militaris in Hirsutella sinensis sta tradicionalno poznani po svojem imunomodulatornem in protivnetnem delovanju, zaradi česar sta vse bolj zanimivi tudi v sodobni medicini, predvsem kot potencialni dopolnili pri zdravljenju bolezni z izraženimi vnetno-fibrotičnimi procesi, kot je sistemska skleroza (SSc). Namen magistrskega dela je zato in vitro ovrednotiti protivnetne in protifibrotične učinke različnih etanolnih ekstraktov teh gliv ter kordicepina, domnevne glavne biološko aktivne spojine. Pri tem nas je zanimal tudi vpliv koncentracije etanola pri pripravi ekstraktov na njihovo sestavo in posledično biološko aktivnost. Raziskavo smo izvedli na dveh celičnih modelih: monocitni liniji THP-1, stimulirani z lipopolisaharidom (LPS), ter primarnih dermalnih fibroblastih (NHDF), stimuliranih s transformirajočim rastnim faktorjem beta (TGF-β). Rezultati so potrdili protivnetni potencial ekstraktov obeh gliv, ki so značilno znižali izražanje IL-6 in IL-1β v LPS-stimuliranih monocitih, pri čemer so se ekstrakti z višjim deležem etanola izkazali kot bolj učinkoviti. Kordicepin samostojno ni pokazal protivnetnega delovanja na celicah THP-1, kar smo pripisali njegovi hitri razgradnji v celicah zaradi encima adenozin deaminaze (ADA); zanimivo pa so glivni ekstrakti, ki naravno vsebujejo inhibitorje ADA (npr. pentostatin), omogočili njegovo učinkovito delovanje. Protifibrotične učinke na fibroblastih smo zabeležili predvsem pri ekstraktih glive H. sinensis in pri kombinaciji kordicepina s pentostatinom, ki so značilno znižali izražanje ključnih fibrotičnih označevalcev (COL1A1, α-SMA in fibronektin) na ravni mRNA in proteinov. Čeprav ekstrakti glive C. militaris niso dosledno pokazali protifibrotičnih učinkov v vseh ponovitvah, rezultati preliminarnih analiz nakazujejo potencial, ki zahteva nadaljnjo potrditev. Pri vrednotenju koncentracijske odvisnosti smo opredelili učinkovite, a netoksične koncentracije, kar je pomemben korak za načrtovanje nadaljnjih študij. Naša raziskava je tako pomembno prispevala k razumevanju kompleksnih protivnetnih in protifibrotičnih mehanizmov kordicepina in glivnih ekstraktov ter izpostavila pomen sočasne prisotnosti inhibitorjev ADA za biološko aktivnost kordicepina. Kljub metodološkim omejitvam in in vitro naravi študije naši rezultati predstavljajo dragoceno osnovo za nadaljnje raziskave in razvoj terapevtskih pristopov za bolezni, kot je sistemska skleroza. The fungi Cordyceps militaris and Hirsutella sinensis are traditionally known for their immunomodulatory and anti-inflammatory properties, attracting growing interest in contemporary medicine as potential therapeutic agents for systemic sclerosis (SSc) and other diseases characterized by inflammatory and fibrotic processes. The aim of this master's thesis was to evaluate the anti-inflammatory and anti-fibrotic effects of various ethanol extracts of these fungi and cordycepin, presumed to be the primary bioactive compound, in vitro. We also investigated how different ethanol concentrations used during extract preparation influenced their biochemical composition and biological activities. Two cellular models were employed: the monocytic THP-1 cell line stimulated with lipopolysaccharide (LPS) and primary dermal fibroblasts (NHDF) stimulated with transforming growth factor-beta (TGF-β). The results confirmed the anti-inflammatory potential of fungal extracts, significantly reducing IL-6 and IL-1β expression in LPS-stimulated monocytes, with extracts prepared using higher ethanol concentrations demonstrating superior efficacy. Cordycepin alone did not exhibit anti-inflammatory activity in monocytes, likely due to rapid degradation by cellular adenosine deaminase (ADA); interestingly, fungal extracts naturally containing ADA inhibitors (such as pentostatin) effectively facilitated cordycepin's biological activity. Anti-fibrotic effects in fibroblasts were notably observed with H. sinensis extracts and the cordycepin-pentostatin combination, significantly decreasing the expression of key fibrotic markers (COL1A1, α-SMA, and fibronectin) at both mRNA and protein levels. Although C. militaris extracts did not consistently demonstrate anti-fibrotic effects across all repetitions, preliminary analyses suggest a promising potential warranting further validation. Dose-response evaluations allowed identification of effective yet non-toxic concentrations, a crucial step for future study design. Our research thus contributes significantly to understanding the complex anti-inflammatory and anti-fibrotic mechanisms of cordycepin and fungal extracts, highlighting the importance of concurrent ADA inhibition for cordycepin’s efficacy. Despite methodological constraints and the in vitro nature of this study, our findings provide a valuable foundation for future research and therapeutic development aimed at inflammatory-fibrotic diseases, such as systemic sclerosis. Cordyceps militaris Hirsutella sinensis kordicepin monociti fibroblasti Cordyceps militaris Hirsutella sinensis cordycepin monocytes fibroblasts true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-10-21 08:45:08 2025-10-21 08:45:24 2025-12-07 03:50:03 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 119597 254386435 29664.pdf 29664.pdf 1 4A58BFCA2AADA59379B9F4FE09491EC6 0e09714a159870c463c3c330043fbc1c4661e0623a4b677b2ac387a98ec87831 3f119722-ae49-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=220501 Fakulteta za farmacijo 0 0 0