Title Učinci ekstrakta crnoplodne aronije na primarne mišje astrocite izložene djelovanju 6-hidroksidopamina
Title (english) Effects of Aronia Melanocarpa extract on primary mouse astrocytes exposed to 6-hydroxydopamine
Author Ela Mijatović
Mentor Kristina Pilipović (mentor)
Committee member Dinko Vitezić (predsjednik povjerenstva)
Committee member Jasenka Mršić-Pelčić (član povjerenstva)
Committee member Kristina Pilipović (član povjerenstva)
Granter University of Rijeka Faculty of Medicine (Department of Pharmacology) Rijeka
Defense date and country 2021-07-15, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Basic Medical Sciences Pharmacology
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Basic Medical Sciences Neuroscience
Abstract Parkinsonova bolest (engl. Parkinson’s disease, PD) najčešći je poremećaj kretanja te druga najučestalija progresivna neurodegenerativnu bolest u svijetu. Korištenjem neurotoksina 6-hidroksidopamin (engl. 6-hydroxydopamine, 6-OHDA) u in vivo i in vitro modelima PD može se doprinijeti rasvjetljavanju patogeneze PD. Terapijske opcije u liječenju PD su limitirane, a sve je veći broj znanstvenih dokaza o neuroprotektivnosti antioksidanasa poput crnoplodne aronije, odgovornih za zaštitu dubokih jezgara mozga poput substantie nigre i bazalnih ganglija. Cilj ovog istraživanja bio je utvrditi učinak tretmana ekstraktom aronije astrocita izloženih djelovanju 6-OHDA.
Pokusi su učinjeni na primarnim mišjim astrocitima koji su bili izloženi tijekom 24 h različitim koncentracijama ekstrakta crnoplodne aronije (10, 50 ili 100 µg/mL). 6-OHDA je dodan u medij za uzgoj stanica tijekom sljedećih 72 h (za test laktatne dehidrogenaze, LDH) ili 24 h (za western blot analize). Razina aktivnosti LDH testirana je radi procjene učinaka tretmana na preživljenje astrocita, a Western blotom detektirana je razina izražaja pojedinih proteina od interesa.
Rezultati su pokazali da tretman različitim koncentracijama ekstrakta aronije ne utječe značajno na aktivnost LDH u uzorcima medija neozlijeđenih astrocita. Uočeno je i da aplikacija 6-OHDA značajno povećava aktivnost LDH, a da su stanice pretretirane ekstraktom aronije u koncentracijama 50 i 100 μg/mL imale značajno nižu razinu aktivnosti ovog enzima. Nadalje, mjerene su razine izražaja proteina toplinskog šoka 70 (engl. heat shock protein 70, HSP70), moždanog neurotrofnog čimbenika (engl. brain derived neurotrophic factor, BDNF) te inducibilne sintaze dušikovog oksida (engl. inducible nitric oxide synthase, iNOS) u astrocita izloženih djelovanju 6-OHDA, netretiranih ili tretiranih ekstraktom aronije. Pretretman aronijom nije značajno utjecao na razine protektivnih proteina HSP70 i BDNF, ali je povećao razinu izražaja iNOS u ozlijeđenim astrocitima, no ne i statistički značajno.
Ova studija ukazuje na moguće protektivne učinke ekstrakta crnoplodne aronije u in vitro modelu PD. Točan mehanizam protektivnog učinka još je uvijek nejasan te bi mogao biti predmetom daljnjih istraživanja.
Abstract (english) Parkinson’s disease (PD) is the most common movement disorder and the second most frequent progressive neurodegenerative disease in the world. Neurotoxin 6-hydroxydopamine (6-OHDA) is used in in vivo and in vitro models of PD and may contribute to elucidating the pathogenesis of this disease. Therapeutic options in the treatment of PD are limited, and there is growing scientific evidence of the neuroprotectiveness of antioxidants such as chokeberry, responsible for protecting deep brain nuclei such as substantia nigra and basal ganglia. The aim of this study was to determine the effect of treatment with chokeberry extract of astrocytes exposed to 6-OHDA.
Experiments were performed on primary mouse astrocytes that were exposed during 24 h to different concentrations of chokeberry extract (10, 50 or 100 µg/mL). 6-OHDA was added to the cell culture medium over the next 72 h (for lactate dehydrogenase test, LDH) or 24 h (for western blot analysis). The level of LDH activity was tested to determine the effects of treatments on astrocyte survival. Western blot analysis detected the level of expression for some proteins of interest.
The results of this study showed that treatment with different concentrations of chokeberry extract did not significantly affect LDH activity in media samples of uninjured astrocytes. Also, it was observed that the application of 6-OHDA significantly increased the activity of LDH, and that the cells treated with chokeberry extract at concentrations of 50 and 100 μg/mL had a significantly lower activity of this enzyme. Furthermore, heat shock protein 70 (HSP70), brain derived neurotrophic factor (BDNF) and inducible nitric oxide synthase (iNOS) expression levels were measured in astrocytes exposed to 6-OHDA, untreated or treated with chokeberry extract. Treatment with chokeberry did not significantly affect the levels of protective proteins HSP70 and BDNF but increased the level of iNOS expression in injured astrocytes, albeit not statistically significantly.
This study suggests possible protective effects of chokeberry extract in an in vitro model of PD. The exact mechanism of the protective effect is still unclear and could be the subject of further research.
Keywords
aronija
astrociti
6-hidroksidopamin
laktat dehidrogenaza
moždani neurotrofni čimbenik
protein toplinskog šoka
Keywords (english)
chokeberry
astrocytes
6-hydroxydopamine
lactate dehydrogenase
brain neurotrophic factor
heat shock protein
Language croatian
URN:NBN urn:nbn:hr:184:337165
Study programme Title: Study of Sanitary Engineering Study programme type: university Study level: graduate Academic / professional title: magistar/magistra sanitarnog inženjerstva (magistar/magistra sanitarnog inženjerstva)
Type of resource Text
File origin Born digital
Access conditions Open access
Terms of use
Created on 2021-06-28 21:06:51