University of Rijeka. Faculty of Medicine. Department of Chemistry and Biochemistry.
Abstract
Uvod: Ulcerozni kolitis (UK) je kronična bolest koja nastaje kao posljedica neodgovarajućeg imunološkog odgovora. Rak debelog crijeva treći je najčešće dijagnosticirani zloćudni tumor i drugi uzrok smrtnosti. Dosadašnje terapije ovih bolesti nisu se pokazale odgovarajućim. Klorogenična kiselina i luteolin su polifenolni spojevi s pokazanim značajnim farmakološkim učincima u tretmanu različitih bolesti.
Cilj istraživanja: Cilj rada je ispitati i predložiti mehanizam djelovanja klorogenične kiseline i luteolina u terapiji UK i raka debelog crijeva.
Materijal i metode: Istraživanje je provedeno na in vivo modelu kolitisa i na in vitro modelu raka debelog crijeva. Tijekom razvoja kolitisa praćene su kliničke promjene, a stupanj oštećenja tkiva debeloga crijeva određen je histopatološkom analizom. Izražaj istraživanih proteina određen je Western blot metodom i imunohistokemijski. Uloga signalnog puta izvanstaničnim signalima reguliranih kinaza (ERK) istražena je primjenom inhibitora. Citotoksičnost ispitivanih spojeva određena je testom stanične vijabilnosti i sposobnosti stvaranja kolonija. Autofagija i apoptoza analizirane su fluorescencijski. Uloga autofagije ispitana je primjenom inhibitora autofagije.
Rezultati: Klorogenična kiselina i luteolin ublažavaju kliničke simptome i smanjuju intenzitet upale. Klorogenična kiselina smanjuje aktivaciju proteina signalnih puteva proteinskih kinaza aktiviranih mitogenom (MAPK), proteinske kinaze B (PKB (Akt)) i pretvarača signala i aktivatora transkripcije 3 (STAT3) te djeluje antioksidacijski. Luteolin aktivira ERK protein. Klorogenična kiselina ne utječe na vijabilnost stanica, dok luteolin smanjuje njihovu vijabilnost i proliferaciju te djeluje proapoptotski, proautofagično i antimetastatski.
Zaključak: Istraživanjem je pokazan protuupalni i antiapoptotski učinak klorogenične kiseline u UK te protuupalni, antiapoptotski i antiautofagični učinak luteolina koji je posredovan ERK signalnim putem. Antikarcinogena aktivnost luteolina se temelji na ERK/FOXO3a ovisnom mehanizmu i antimetastatskom potencijaluIntroduction: Ulcerative colitis (UC) is a chronic disease that results from an inadequate immune response. Colon cancer is the third most commonly diagnosed malignancy and the second leading cause of death. Previous therapies for these diseases have not been shown to be adequate. Chlorogenic acid and luteolin are polyphenolic compounds with a shown significant pharmacological effects in the treatment of various diseases.
Aim of the study: The aim of this study was to investigate and propose the mechanism of action of chlorogenic acid and luteolin in the treatment of UC and colon cancer.
Matherial and methods: The study was performed in an in vivo model of colitis and in an in vitro model of colon cancer. During the development of colitis, clinical changes were monitored and the degree colon tissue damage was determined by histopathological analysis. The expression of the investigated proteins was determined by the Western blot method and immunohistochemically. The role of the extracellular regulated kinases (ERK) signaling pathway was investigated using inhibitor. The cytotoxicity of the test compounds was determined by a cell viability and colony formation assay. Autophagy and apoptosis were analyzed by fluorescence. The role of autophagy was investigated using autophagy inhibitor.
Results: Chlorogenic acid and luteolin alleviate clinical symptoms and reduce the intensity of inflammation. Chlorogenic acid reduces the activation of mitogen-activated protein kinases (MAPK), protein kinase B (PKB (Akt)) and signal transducer and transcription activator 3 (STAT3) signaling pathway proteins and has an antioxidant effect. Luteolin activates ERK protein. Chlorogenic acid does not affect cell viability, while luteolin reduces their viability and proliferation and has proaptotic, proautophagic and antimetastatic effect.
Conclusion: The study demonstrated the antiinflammatory and antiapoptotic effect of chlorogenic acid in the UC and the antiinflammatory, antiapoptotic and antiautophagic effect of luteolin mediated by the ERK signaling pathway. The anticarcinogenic activity of luteolin is based on the ERK/FOXO3a dependent mechanism and antimetastatic potential
