| Abstract | UVOD: Ulcerozni kolitis (UK) je idiopatska kronična upalna bolest gastrointestinalnog sustava praćena mnogim komplikacijama. Upala zahvaća sluznicu debelog crijeva i to najčešće završni dio debelog crijeva, a etiopatogeneza nije u potpunosti razjašnjena. Istraživanja ukazuju kako upala nastaje u interakciji niza čimbenika uključujući genetsku predispoziciju, imunološki sustav, mikroorganizme i okoliš. U organizmu, morfološki razlikujemo tri vrste stanične smrti, jedna od njih je apoptoza ili programirana stanična smrt. Apoptoza je proces koji sudjeluje u regulaciji održavanja tkivne homeostaze, a njezini poremećaj može dovesti razvoja mnogih oboljenja ukjučujući imunološke i degenerativne te tumora. Pokazano je da stupanj apoptoze, kako upalnih tako i epitelnih stanica, utječe na brzinu razvoja i intenzitet upalnog procesa. Brojna istraživanja ukazuju da hrana bogata klorogenim kiselinama pogoduje ljudskom zdravlju te da imaju antioksidativni i protuupalni učinak.
CILJ: Cilj predloženog istraživanja je ispitati izražaj pro- i protuapoptotskih molekula u kolitisu te utjecaj klorogenične kiseline (KGK) na razvoj upale i aktivaciju procesa apoptoze.
MATERIJALI I METODE: U sklopu provedbe istraživanja korišteni su C57BL/6 miševi u kojih je primjenom 2,5% otopine natrijevog dekstran sulfata (DSS) kroz sedam dana izazvan pokusni model kolitisa. Procjena uspostave i intenziteta kolitisa praćena je na dnevnoj bazi putem pojavnosti i intenziteta markera upalnog procesa te izračunom indeksom aktivnosti bolesti. Na proteinskoj razini, u tkivu debeloga crijeva, pratila se promjena izražaja pro- i protuapoptotskih molekula.
REZULTATI: Tijekom primjene DSS otopine zabilježen je pad tjelesne mase, porast indeksa aktivnosti bolesti te oštećenje sluznice kod svih ispitivanih skupina životinja. Međutim, u skupini miševa kod kojih se je uz DSS primijenila KGK gubitak tjelesne mase značajno je manji, a također i indeks aktivnosti bolesti. U upalno promijenjenom tkivu životinja kod kojih je primjenjena samo DSS otopina, u odnosu na kontrolnu skupinu, pojačan je izražaj Bax, kaspase 9, cijepane kaspaze 8, a smanjen izražaj Bcl-2 i kaspaze 8. Primjena KGK dovodi do obrata izražaj istih, a stupanj promjene ovisan je o primjenjenoj dozi.
ZAKLJUČAK: Primjena KGK značajno utječe na intenzitet kolitisa te na izražaj pro- i protuapoptotskih molekula. Stoga, rezultati studije ukazuju da bi se KGK mogla koristiti u terapiji upalom posredovanih bolesti.
Ključne riječi: Ulcerozni kolitis, apoptoza, klorogenična kiselina, animalni modeli. |
| Abstract (english) | INTRODUCTION: Ulcerative colitis (UC) is an idiopathic chronic inflammation of gastrointestinal system, accompanied with wide variety of extraintestinal manifestations. Inflammation affects colon's mucous, most often final portion of the large intestine and etiopathogenesis of inflammation is still not fully understood. There are 3 types of cell death in multicellular organism, one of them is being apoptosis well known as programmed cell death. Apoptosis plays a fundamental role in human development and tissue homeostasis. Aberrant regulation of apoptosis is linked with a various disease, including immunological and developmental disorders, and cancer. According to vast number of studies, food rich with chlorogenic acids favours human health and has antioxidative and anti-inflammatory impact.
AIM: The aim of the proposed study was to investigate the expression of pro- and anti-apoptotic molecules in DSS colitis and the influence of chlorogenic acid (CHA) on the development of inflammation and the activation of the apoptosis.
MATERIALS AND METHODS: In the current study male C57BL/6 mice were used. Colitis was induced by by administration of 2,5%. DSS in driking water for seven days. Evaluation of colitis development and severity was monitored daily through occurence and intensity of clinical markers and by calculation of disease activity index. Analysis of results established disease activity indeks. At the protein level, changes in the expression of pro- and anti-apoptotic proteins were observed in the colon tissue.
RESULTS: During the administration of the DSS solution, a decrease in body weight, and increase in the disease activity index, and mucosal damage in all groups of animals were observed. However, CHA supplementation attenuated weight loss significantly and conequently disese activity index was lower. A increased expression of Bax, caspase 9, cleaved caspase 8 and decreased expression of Bcl-2 and caspase 8 was determined in mice treated only with DSS in comparison to the control group. Treatment with CHA dose-dependently influenced on the expression of proapoptotic and antiapoptotic molecules.
CONCLUSION: The appliance of KGK significanlty affects colitis development as well as the expression of pro and antiapoptosis molecules. The results of the study suggest that CHA could be usful for the tretment of inflammation. |
