Preizkušanje učinkovitosti peptidnega antagonista grelinskega receptorja na mišjem modelu : Industrijska farmacija

Peinkiher, Sara (author), Lunder, Mojca (mentor), Grgurevič, Neža (comentor)

Grelin je periferni peptidni hormon, ki je vpleten v uravnavanje energijske homeostaze, uravnavanje vnosa hrane in izločanje rastnega hormona. Iskanje antagonistov receptorja za grelin je eden od načinov iskanja novih spojin vodnic, katerih namen je zdravljenje bolezenskih stanj in tveganj, kot so na primer prekomerna telesna masa, debelost in z njo povezana druga bolezenska stanja. Na Katedri za farmacevtsko biologijo, Fakultete za farmacijo, Univerze v Ljubljani, so s pomočjo tehnologije bakteriofagnega prikaza razvili peptidne ligande receptorja za grelin, z in vitro antagonističnim delovanjem. Najučinkovitejšega med njimi smo v magistrski nalogi preizkusili še in vivo na mišjem modelu linije C57BL/6J. Delovanje peptida, ki smo ga poimenovali P1, smo primerjali z antagonistom grelina [D-Lys-3]-GHRP-6. V nalogi smo preizkusili dva načina vnosa snovi, preko trebušne (intraperitonealno, i.p.) in preko nosne votline. Opravili smo 3 poskuse; v poskusih 1 in 2 smo po vnašanju snovi intraperitonealno in preko nosne votline spremljali vnos hrane pri testnih živalih, v poskusu 3 smo po vnosu peptidne snovi intraperitonealno merili koncentracijo rastnega hormona v serumu testnih živali. V poskusih 1 in 2 smo imeli tri skupine živali: 1) kontrolno, 2) skupino z dodanim [D-Lys-3]-GHRP-6 in 3) skupino z dodanim peptidom P1. V poskusu 3 smo poleg v poskusih 1 in 2 navedenih treh dodali še tri skupine živali: 4) z dodanim grelinom, 5) z istočasno dodanim grelinom in peptidom P1 ter 6) z istočasno dodanim [D-Lys-3]-GHRP-6 in peptidom P1. Prost vnos hrane 30 min po intraperitonealni aplikaciji snovi se je razlikoval med kontrolno skupino in skupino z dodanim antagonistom [D-Lys-3]-GHRP-6, medtem ko pri skupini z dodanim peptidom P1 ni prišlo do pričakovanih sprememb. Razlike smo zaznali tudi v koncentraciji rastnega hormona v serumu. Aplikacija grelina je, v primerjavi z nosilnim sistemom brez grelina, signifikantno povečala koncentracijo rastnega hormona v serumu. Istočasna aplikacija grelina in peptida P1 je povzročila nepričakovano višjo koncentracijo rastnega hormona kakor sama aplikacija grelina. Rezultati naše raziskave niso potrdili antagonističnega in vivo delovanja peptida P1 na vnos hrane, pokazali pa so vpliv peptida P1 na izločanje rastnega hormona.

• Language: Slovenian
• Keywords: peptid P1, antagonisti, grelin, in vivo testiranje, mišji model
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Place of publishing: Ljubljana
• Publisher: [S. Peinkiher]
• Year: 2018
• Number of pages: VIII, 37 f.
• PID: 20.500.12556/RUL-120272
• UDC: 543.645.6(043.3)
• COBISS.SI-ID: 4462193
• Publication date in RUL: 17.09.2020

Secondary language

• Language: English
• Title: Peptide of ghrelin receptor efficacy in the mouse model

Abstract

Ghrelin is a peripheral peptide hormone, which is involved in the regulation of energy homeostasis, regulation of food intake and secretion of growth hormone. Searching for the ghrelin receptor antagonist is one way of finding a new lead compounds in order to treat conditions, such as overweight, obesity and its other related illness conditions. Department of Pharmaceutical Biology, Faculty of Pharmacy of University of Ljubljana has developed new peptide ligands of the ghrelin receptor with the help of bacteriophage imaging technology, which have expressed in vitro antagonistic activity. In the master’s thesis we have tested the most effective peptide ligand among them (peptide P1) in vivo study on the C57BL/6J mouse model strain. The activity of peptide, which we have named P1, was compared with the known ghrelin antagonist [D-Lys-3] -GHRP-6. In our thesis we have tested two methods of applying substances, through abdominal and nasal cavity. We conducted 3 experiments; in experiments 1 and 2 we have monitored the food intake depending on the different application of substances. In the experiment 3, we have monitored the concentration of growth hormone in the serum of the tested animals after intraperitoneal application. In experiments 1 and 2 we had three testing groups of animals; 1) a control group, 2) group with the addition of [D-Lys-3]-GHRP-6 and 3) the group with added peptide P1. In experiment 3, additional three testing groups were added to the experiment: 4) group with the addition of ghrelin, 5) with simultaneously appliedghrelin and peptide P1 and 6) with simultaneously applied [D-Lys-3]-GHRP-6 and peptide P1. Ad libitum food intake after 30 min from intraperitoneal application was distinguished between control group and group with [D-Lys-3] - GHRP-6, while no expected changes were noticed in the group with added peptide P1. Differences were also observed in the concentration of serum growth hormone. The application of ghrelin has, in comparison with the carrier (control group), significantly increased expression of growth hormone in the serum. Simultaneous application of ghrelin and peptide P1 resulted in an unexpectedly higher growth hormone concentration than the application of ghrelin itself. The results of our study did not confirm the antagonistic in vivo activity of peptide P1 on the food intake, but they showed effect of peptide P1 on the secretion of growth hormone release.

• Keywords: peptide P1 peptides antagonist ghrelin in vivo testing mouse model