Vpliv olanzapina na koncentracijo katalitične aktivnosti ALP v živalskem modelu podgan z izbitim genom ETBR

Malešič, Saša (author), Marc, Janja (mentor), Prodan Žitnik, Irena (co-mentor)

Dokazano je, da endotelinski sistem preko celičnih signalnih poti aktivira osteoblastogenezo in s tem kostno tvorbo. Namen magistrske naloge je bil ugotoviti, ali ti učinki potekajo preko beta endotelinskega receptorja (ETB) in ali na te učinke vpliva jemanje antipsihotika olanzapina. Raziskava je bila izvedena na živalskem modelu podgan z izbitim genom za endotelinski receptor (gen ETBR). Pri testnih skupinah podgan je bila kostna premena pospešena z aplikacijo superelastične vzmeti (ortodontski premik zob).Vključenih je bilo 98 podgan, razdeljenih v skupine glede na prisotnost gena ETBR, prejemanje olanzapina in nameščenost ortodontske vzmeti (aktivni premik zob). V serumih podgan smo merili koncentracijo katalitične aktivnosti alkalne fosfataze (ALP), ki je biokemijski označevalec kostne tvorbe. Uporabili smo referenčno kinetično IFCC merilno metodo, katere uporabnost je zelo omejena v primeru prisotne hemolize v vzorcih seruma. S spremembo valovne dolžine, pri kateri se meri absorbanca nastalega produkta, smo uspeli izmeriti katalitično koncentracijo tudi v hemoliziranih vzorcih. Ustreznost živalskega modela smo preverili z merjenjem serumske koncentracije endotelina-1 (ET-1). Slednjo smo merili z uporabo ELISA testa. Koncentracija ET-1 je bila značilno višja v vseh skupinah podgan, ki so imele izbit gen za receptor ETB (ETBR). S tem smo potrdili, da je ETBR odgovoren za klirens ET-1 iz krvnega obtoka. Med podganami z izbitim genom ETBR in nativnimi podganami nismo ugotovili značilne razlike v katalitični aktivnosti ALP. Znotraj posamezne populacije podgan pa je bila katalitična aktivnost ALP značilno višja v skupinah, ki so prejemale olanzapin, kot v skupinah, ki olanzapina niso prejemale, ne glede na prisotnost gena ETB R. Rezultati naloge kažejo, da olanzapin zviša kostno tvorbo pri podganah z nameščeno superelastično vzmetjo (ortodontski premik zob), ter da se vpliv endotelina na kostno tvorbo poveča v odsotnosti endotelinskega receptorja beta.

• Language: Slovenian
• Keywords: kostna tvorba, endotelinski receptor beta, olanzapin, alkalna fosfataza
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2021
• PID: 20.500.12556/RUL-127918
• Publication date in RUL: 29.06.2021

Secondary language

• Language: English
• Title: Olanzapine impact on ALP catalytic activity concentration in the ETBR gene knocked-out rat animal model

Abstract

The endothelin peptide family promotes osteoblastogenesis and stimulaties bone formation by activating cellular signalling pathways. The aim of this thesis was to determine whether this process is influenced by the endothelin receptor beta (ETB) and the presence of antipsychotic olanzapine. An ETBR gene knockout rat animal model was used in the research. Bone remodelation was accelerated in the experimental groups by the presence of a super elastic coiled spring (orthodontic tooth movement). The research included 98 rats divided into subgroups depending on the presence of the ETBR gene, administration of olanzapine and the placing of the coiled spring between two of the rats molars. The catalytic activity concentrations of alkaline phosphatase (ALP) were measured in rat serums as a bone formation marker. The IFCC kinetic alkaline phosphatase method was used for measuring ALP. The application of the latter turned out to be very limited in the presence of hemolysis in serum samples. We therefore attempted to diminish the interference of hemoglobine by changing the wavelength used to measure the formation of the reaction product 4-nitrophenol. The suitability of the animal model was verified by measuring endothelin-1 (ET-1) concentrations in rat serums. This was measured using an ELISA test. The concentration of ET-1 was significantly higher all ETBR gene knockout groups when compared to the native groups. This result confimed that the ETB receptor is responsible for the majority of ET-1 clearance from the blood stream. There was no significant difference in the ALP catalytic activity between the native and mutant rat populations. However, within populations, the catalytic activity concentration of ALP was significantly higher in groups (ETBR knockout and native) in which olanzapine was administrated, compared to those where it was not, regardless of the presence of the ETBR gene. The results show that olanzapine increases the rate of bone formation in rats that are undergoing active orthodontic tooth movement. It also amplifies bone formation in the absence of ETBR.

• Keywords: bone formation, endothelin receptor B, olanzapine, alkaline phosphatase