Povezava kazalnikov oksidativnega stresa z aktivnostjo kronične vnetne črevesne bolezni pri zdravljenju z ustekinumabom

Kralj, Manca (author), Vovk, Tomaž (mentor), Tratenšek, Armando (comentor)

Incidenca in prevalenca kronične vnetne črevesne bolezni rasteta. Diagnoza bolezni je pogosto zakasnjena, saj so simptomi nespecifični, s tem pa je povezana slabša kakovost življenja bolnikov in večja možnost zapletov. Trenutno uporabljene diagnostične metode so invazivne in drage, s čimer narašča potreba po odkritju novih bioloških kazalnikov in razvoju novih metod. Za izbrane kazalnike oksidativnega stresa smo preverili, ali se njihove vrednosti razlikujejo med bolniki v aktivni fazi in remisiji. Za določanje prostih tiolov in naprednih oksidiranih proteinskih produktov smo uporabili neseparacijske spektrofotometrične metode, malondialdehid pa smo, po predhodni derivatizaciji z dinitrofenilhidrazinom, določili s separacijsko metodo tekočinske kromatografije visoke ločljivosti z ultravijoličnim detektorjem. Metodi določanja naprednih oksidiranih proteinskih produktov in prostih tiolov smo delno validirali. V območju od 10-100 µmol/L za proste tiole in 100-900 µmol/L za napredne oksidirane proteinske produkte nam metodi dajeta točne in ponovljive rezultate. Zaradi omejene količine biološkega vzorca smo metodo določanja malondialdehida optimizirali za manjše volumne. V raziskavo je bilo vključenih 49 bolnikov s kronično vnetno črevesno boleznijo na terapiji z ustekinumabom, vzorci pa so bili odvzeti v dveh časovnih točkah (ob času 0 in 24 tednov). Na podlagi endoskopske aktivnosti bolezni, določene z indeksom SES-CD ter s fekalnim kalprotektinom, smo primerjali vrednosti kazalnikov pri bolnikih z aktivno boleznijo ter tistih v remisiji. Vrednosti malondialdehida in oksidiranih proteinskih produktov so bile višje v plazmi bolnikov z aktivno kronično vnetno črevesno boleznijo kot pri tistih v remisiji, vendar razlike niso bile statistično značilne. Med skupinama nismo zaznali razlik v vrednostih prostih tiolov. Opazili smo zelo šibko pozitivno korelacijo med fekalnim kalprotektinom in C-reaktivnim proteinom z malondialdehidom in naprednimi oksidiranimi proteinskimi produkti. Na podlagi naših rezultatov je diagnostična moč testov za napredne oksidirane proteinske produkte, proste tiole in malondialdehid prenizka za uporabo v kliničnem okolju. Glede na opažen trend v razlikah v koncentracijah med aktivno in neaktivno fazo bolezni so smiselne nadaljnje raziskave o povezavi kazalnikov oksidativnega stresa in aktivnosti kronične vnetne črevesne bolezni.

• Language: Slovenian
• Keywords: kronična vnetna črevesna bolezen, Crohnova bolezen, ulcerozni kolitis, kazalniki oksidativnega stresa, ustekinumab
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2022
• PID: 20.500.12556/RUL-138049
• Publication date in RUL: 08.07.2022

Secondary language

• Language: English
• Title: Correlation of oxidative stress markers with inflammatory bowel disease activity in ustekinumab treated patients

Abstract

The incidence and prevalence of chronic inflammatory bowel disease are increasing. Diagnosis of the disease is often delayed because of the nonspecific symptoms, resulting in poorer quality of life and increased risk of complications. Currently used diagnostic methods are invasive and expensive, thus the discovery of new biological markers and the development of new methods are urgently needed. We investigated whether the values of selected markers differ between patients in the active phase of the disease and those in remission. The free thiols and advanced oxidation protein products were determined using non-separation spectrophotometric methods, and the concentration of malondialdehyde was measured by high-performance liquid chromatography with ultraviolet detector after prior derivatization with dinitrophenylhydrazine. Methods for the determination of advanced oxidation protein products and free thiols were partially validated. In the range of 10-100 µmol/L for free thiols and 100-900 µmol/L for advanced oxidation protein products, the methods provide accurate and reproducible results. The method for the determination of malondialdehyde was optimized for smaller volumes due to the limited amount of biological sample. The study included 49 patients with chronic inflammatory bowel disease treated with ustekinumab, whose samples were collected at two different time points (after 0 and 24 weeks). Based on the endoscopic activity of the disease determined by the SES-CD index and fecal calprotectin, we compared the values of markers in patients with active disease and those in remission. The levels of malondialdehyde and advanced oxidation protein products in plasma of patients with active chronic inflammatory bowel disease were higher than in those in remission, but the differences were not statistically significant. There were no differences in free thiol values between the groups. A very weak positive correlation was observed between fecal calprotectin, C-reactive protein and markers malondialdehyde and advanced oxidation protein products. According to our results, the diagnostic power of markers advanced oxidation protein products, free thiols and malondialdehyde is insufficient to be used in the clinical settings. Given the observed trend in concentration differences between groups, further research is needed to evaluate the relationship between oxidative stress markers and chronic inflammatory bowel disease activity.

• Keywords: inflammatory bowel disease, Crohn's disease, ulcerative colitis, oxidative stress markers, ustekinumab