Periferni i centralni učinci botulinum toksina tipa A kod imunogene upale koljena štakora izazvane Freundovim adjuvansom

master's thesis

• Title (english): Peripheral and central effects of botulinum toxin type A in Freund's adjuvant- induced immunogenic inflammation in rat knee
• Author: Mario Špoljarić
• Mentor: Ivica Matak (mentor)
• Committee member: Ante Tvrdeić (predsjednik povjerenstva)
• Committee member: Jasenka Markeljević (član povjerenstva)
• Committee member: Ivica Matak (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Pharmacology); Zagreb
• Defense date and country: 2019-07-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Pharmacology

Abstract

Uvod i cilj: Botulinum neurotoksin tip A (BoNT/A) selektivno ulazi u neurone i inhibira egzocitozu neurotransmitera u sinaptičku pukotinu proteolitičkim cijepanjem sinaptosomalnog proteina od 25 kilodaltona (kDa). Cilj ovoga pokusa bio je istražiti periferne i središnje protuupalne i antinociceptivne učinke BoNT/A kod upale koljena štakora izazvane kompletnim Freundovim adjuvansom (CFA). ----- Materijali i metode: BoNT/A je apliciran intratekalno (i.t.) na razini lumbalne kralježnice (2 U/kg/10μl), ili intraartikularno (i.a.) u koljeno (7 U/kg/20μl). Nakon 7 dana, izazvana je upala koljena injekcijom CFA (50 μl). Za evaluaciju edema životinjama je mjeren promjer koljena, a nociceptivni bihevioralni odgovor pomoću uređaja za mjerenje primijenjenog pritiska (engl. Pressure Application Measurement-PAM), prije i poslije tretmana s CFA. Dva dana nakon indukcije upale, učinak na ekstravazaciju proteina plazme je određen spektrofotometrijski mjerenjem sadržaja kompleksa Evans blue i proteina plazme u uklonjenom tkivu zgloba.
----- Rezultati: BoNT/A injiciran direktno u zglob (i.a.) je smanjio promjer natečenog zgloba s maksimalnim učinkom dva dana nakon početka upale, dok BoNT/A primijenjen i.t. nije smanjio edem. S druge strane, učinak na smanjenje ekstravazacije je bio statistički značajan samo u skupini kojoj je BoNT/A primijenjen u lumbalni intratekalni prostor.
----- Zaključak: Botulinum toksin smanjuje oteklinu zgloba samo nakon periferne primjene, a ekstravazaciju proteina nakon središnje primjene. Uzrok tome bi moglo biti djelovanje na različite medijatore imunogene upale ovisno o mjestu primjene, a za otkrivanje molekularne podloge različitog učinka na pojedine komponente upale potrebna su daljnja istraživanja.

Abstract (english)

Introduction and aim: Botulinum neurotoxin type A (BoNT/A) selectively enters neurons and inhibits synaptic neurotransmitter exocytosis by proteolytic cleavage of synaptosomal-associated protein 25 (SNAP-25). The aim of this experiment was to evaluate peripheral and central anti-inflammatory and antinociceptive effects of BoNT/A in rats with complete Freund's adjuvant (CFA)- induced knee joint inflammation.
----- Materials and methods: Botulinum toxin was administered intrathecally (i.t.) at the level of lumbar spine (2 U/kg/10μl), or intraarticularly (i.a.) into the rat knee (7 U/kg/20μl). After 7 days, to induce inflammation, animals were injected with CFA (50 μl). The knee diameter was measured for evaluation of edema, while nociceptive behavioural response was assessed by employing the Pressure Application Measurement (PAM) device . The effect on plasma protein extravasation was evaluated using spectrophotometry by measuring the removed joint tissue content of Evans-blue- plasma protein complex , two days after induction of the inflammation.
----- Results: BoNT/A injected directly into the joint (i.a.) significantly reduced edema, with the peak effect on the second day of the inflammation. BoNT/A administered intrathecally didn't reduce edema. On the other hand, the significant reduction of plasma protein extravasation was observed only in BoNT/A i.t- injected animals.
----- Conclusion: In present experiments, BoNT/A reduces edema only when delivered in the peripheral nervous system, while it reduces plasma protein extravasation when administered into the CNS. Dependently on the toxin application site, differences in observed BoNT/A effects might be caused by different inflammatory mediators affected. Further experiments to determine the molecular mechanisms of BoNT/A anti-inflammatory action are needed.

• Keywords: botulinum toksin tipa A; upala; kompletni Freundov adjuvant; edem; ektravazacija
• Keywords (english): botulinum toxin; inflammation; complete Freund's adjuvant; edema; extravasation
• Language: croatian
• URN:NBN: urn:nbn:hr:105:049087
• Study programme: Title: Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2019-12-18 15:07:00