Vpliv bosvelične kisline na vnetni odziv celic Caco2, stimuliran z lipopolisaharidom

Kukman, Živa (avtor), Štrukelj, Borut (mentor), Jamnik, Polona (komentor)

Z vedno večjo pojavnostjo kroničnih vnetnih bolezni (revmatoidni artritis, ulcerozni kolitis, spondiloartritis, luskavica, Crohnova bolezen ...) ter doživljenjskim načinom zdravljenja z zdravili, ki povzročajo veliko neželenih učinkov, se povečuje tudi zanimanje za naravne protivnetne učinkovine. V Sloveniji manj poznana, a drugod že dobro uveljavljena protivnetna zdravilna rastlina, je tudi Boswellia serrata. Bosvelija proizvaja smolo, katera vsebuje acetil-11-keto-β-bosvelično kislino. Ta je je nosilka njenega protivnetnega delovanja. Namen našega eksperimentalnega dela je bil podrobneje razumeti delovanje bosvelične kisline na molekularni ravni in s tem določiti tarčno skupino obolenj za zdravljenje z ekstraktom bosvelije. Z namenom pridobitve celotne slike dogajanja smo izbrali 2-D elektroforezo kot analizno metodo. Celicam Caco-2 smo dodali najprej lipopolisaharid (LPS) kot sprožilec vnetja. Nato smo tem istim celicam z LPS-om, dodali še ekstrakt bosvelije. Celične lizate smo prenesli v laboratorij na Biotehniško fakulteto, kje smo jih razvili z 2-D elektroforezo. Slike smo primerjali s programom 2-D Dymension, kjer smo iskali proteinske lise z različno intenziteto. Našli smo 4 lise, ki so bile pri celicah z dodanim LPS, glede na same celice, bolj intenzivne. Ob primerjanju istih lis pri slikah celic z dodano bosvelijo, v primerjavi s samim LPS, pa se je njihova intenziteta zmanjšala. Izbrane proteinske lise smo poslali na Inštitut Ruđer Boškovič, kjer so proteine identificirali. Rezultati so pokazali, da bosvelija deluje zaviralno na protein kalmodulin. Kalmodulin je kalcij vezavni protein in sodeluje v telesu pri mnogih fizioloških procesih, eno izmed področji je tudi vnetje. Vključuje se v proces nastanka IL-6, TNF-α, IL-1β, IL-2; hkrati pa bi lahko potencialno deloval kot RAF-1 inhibitor, ki zavira proliferacijo rakavih celic. Verjetno je še več farmakoloških vplivov izvlečka bosvelije, zato so poti njenega delovanja odprte za nadaljnja raziskovanja. Glede na varnost naravne zdravilne rastline Boswellia serrata, kar smo preverili s testom metabolične aktivnosti, bi lahko bila potencialno registrirana kot zdravilo rastlinskega izvora in bi se uporabljala pri lažjih oblikah kroničnih vnetnih bolezni.

• Jezik: Slovenski jezik
• Ključne besede: Kronično vnetje, Boswellia serrata, 2-D elektroforeza, vnetni proteini, kalmodulin
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2021
• PID: 20.500.12556/RUL-124721
• Datum objave v RUL: 12.02.2021

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Influence of boswellic acid on the inflammatory response of the lipopolysaccharide-stimulated Caco2 cells

Izvleček

With the increasing incidence of chronic inflammatory diseases (rheumatoid arthritis, ulcerative colitis, spondyloarthritis, psoriasis, Crohn's disease ...) and lifelong treatment with drugs that cause many side effects, there is also increasing interest in natural anti-inflammatory drugs. Boswellia serrata is also a lesser-known in Slovenia, but well-established anti-inflammatory medicinal plant elsewhere. Boswellia produces a resin that contains acetyl-11-keto-β-boswellic acid, which is the carrier of its anti-inflammatory action. The purpose of our experimental work was to understand in more detail the action of boswellic acid at molecular level and thus to determine the target group of diseases for treatment with boswellia extract. In order to obtain a complete picture of events, we chose 2-D electrophoresis as an analytical method. To Caco-2 cells, lipopolysaccharide (LPS) was added first as an inflammatory trigger. Boswellia extract was then added to these same cells with LPS. The cell lysates were transferred to the laboratory at the Biotechnical Faculty, where 2-D electrophoresis was performed. Images of 2-D gels were compared using 2-D Dymension program to look for over and under-expressed protein spots. We found four spots that were overexpressed in the cells with added LPS, relative to the cells themselves. When comparing the same proteinsin the images of cells with added boswellia, compared to the LPS itself, they were downregulated. Selected protein spots were sent to the Ruđer Boškovič Institute, where the proteins were identified. The results showed that boswellia had an inhibitory effect on the calmodulin. Calmodulin is a calcium binding protein and is involved in many physiological processes in the body, one of the areas in inflammation. It is involved in the process of formation of IL-6, TNF-α, IL-1β, IL-2; at the same time, it could potentially act as an RAF-1 inhibitor that inhibits cancer cell proliferation. There are probably even more pharmacological effects of boswellia extract, so the pathways of its action are open for further research. Given the safety of the natural medicinal plant Boswellia serrata, which was verified by a metabolic activity test, it could potentially be officially recognized as a medicinal plant and used in milder forms of chronic inflammatory diseases with more intensive research.

• Ključne besede: Chronic inflammation, Boswellia serrata, 2-D electrophoresis, inflammatory proteins, calmodulin