Imunohistokemijska detekcija botulinum toksina u spinalnim regijama nakon supraspinalne primjene

master's thesis

• Title (english): Immunohystochemical detection of botulinum toxin in spinal areas after supraspinal application
• Author: Tin Šandl
• Mentor: Lidija Bach-Rojecky (mentor)
• Committee member: Lidija Bach-Rojecky (predsjednik povjerenstva)
• Committee member: Višnja Drinovac (član povjerenstva)
• Committee member: Maja Ortner Hadžiabdić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmacology); Zagreb
• Defense date and country: 2018-04-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Botulinum toksin tipa A (BT-A) indiciran je u poremećajima prvenstveno vezanima uz povećanu spastičnost mišićnih struktura, no fokus najnovijih ispitivanja učinak je neurotoksina na bol. BT-A pokazuje iznimno povoljan farmakoterapijski profil s dugotrajnim antinociceptivnim djelovanjem i malim brojem nuspojava, što ga razlikuje od svih odobrenih analgetika. Smatralo se da BT-A iskazuje antinociceptivni učinak inhibirajući otpuštanje proupalnih medijatora na perifernim okončinama senzornih živčanih vlakana, no pozitivni rezultati u otklanjanju boli upalnoga i neuropatskog porijekla ukazuju na središnje mjesto djelovanja BT-A. Bihevioralni rezultati dobiveni modelima visceralne i zrcalne boli, u kojima je uočen bilateralni analgetski učinak BT-A nakon unilateralne periferne i intratekalne, ali ne središnje intracerebroventrikularne i intracisternalne primjene, poslužili su kao temelj hipoteze o središnjemu segmentalnom analgetskom djelovanju BT-A na spinalnoj razini. Imunohistokemijski je detektirana prisutnost pocijepanoga SNAP-25 proteina (indirektni marker aktivnosti BT-A) predominantno u dorzalnim rogovima segmenata leđne moždine, ali primjenom blokatora aksonalnog transporta prisutnost nije uočena, što pretpostavlja da BT-A nakon retrogradnog aksonalnog transporta s periferije iskazuje antinociceptivni učinak modulirajući prijenos bolnog impulsa senzornim vlaknima na razini prve sinapse u leđnoj moždini.
Ispitivano je ponašanje BT-A nakon supraspinalne primjene u lateralne moždane komore (i.c.v.) i cisternu magnu (i.c.). Iako je pocijepani SNAP-25 detektiran u daljim regijama mozga nakon i.c.v primjene BT-A, broj c-Fos (marker nociceptivne aktivnosti) pozitivnih neurona nije se smanjio. Nakon i.c. primjene BT-A uočena je prisutnost pocijepanoga SNAP-25 u pojedinim vlaknima dorzalnih rogova leđne moždine, no izostao je analgetski učinak.
Smatra se da BT-A djeluje na bol inhibicijom otpuštanja proupalnih medijatora iz okončina senzornih vlakana na razini prve sinapse u leđnoj moždini, kao i interakcijom s neurotransmisijom putem endogenog inhibitornog GABA i opioidnog sustava, što rezultira modulacijom prijenosa bolnog impulsa iz periferije prema središnjim centrima.

Abstract (english)

Botulinum toxine of Ty A (BT-A) is indicated in disorders primary connected to increased spastics of muscular
structures, but the focus of the most recent research is the effect of the neurotoxin to pain. The BT A presents an
extremely favourable pharmaco therapeutic profile with long lasting antinociceptive action and small number of side
effects, making it different from other appr oved analgetics. It was deemed that the BT A shows its antinocicptive
effects by inhibiting the release of anti inflammatory mediators at the peripheral ends of the sensor nerve fibres, but
the positive outcomes in pain killing of inflammatory and neuropat hic origin indicate to the central place of the BT A
action. The behavioral results obtained by models of visceral and mirror pain, in which bilateral analgesic effect of the
BT A after unilateral peripheral and intraecal, but not central intracerebroventi cular and intracisternal application has
been observed, have served as a fondation of the hypothesis on central segmental analgesic action of the BT A at
spinal level. Immunohystochemically, the presence of a cracked SNAP 25 protein (indirect market of BT A activity)
has been detected, predominantly in dorsal spinous processes of the back spinal cord segments. However, by applying
axonal transport blocker, no presence has been observed, assuming that the BT A after the axonal back transport from
the periphe ry is showing an anticiceptive effect by modulating the transport of the painful impulse by means of sensor
fibres at the first synapsis level in the dorsal spine.
The behaving of BT-A after supraspinal application to lateral brain chambers (i.c.v.) and c isterna magna (i.c.) has been
tested. Although the cracked SNAP 25 was detected in further regions of the brain after the i.c.v. application of the
BT a, the number of c Fos (the nociceptive activity marker) positive neurons has not decreased. After the i. c.
application of BT A, the presence of cracked SNAP 25 in particular fibres of dorsal spinous processes of the back
spinal cord has been observed, but the analgesic effect was absent.
It is deemed that the BT-A performs action against pain by inhibiting t he release of anti inflammatory mediators from
the ends of sensor fibres at the first synapsis level in the spinal cord, as well as interacting with the neuro transmission
by means of endogenic GABA and opinoide system, resulting in modulation of the pain impulse transmission from the
periphery to centres.

• Keywords: botulinum toksin; bol; SNAP-25; spinalni segmenti; supraspinalna primjena; imunohistokemija
• Keywords (english): botulinum toxin; pain; SNAP-25; spinal segments; supraspinal application; immunohystochemistry
• Language: croatian
• URN:NBN: urn:nbn:hr:163:162990
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-04-19 12:13:25