Vpliv 6-hidroksidopamina na nivo proteina katepsina X v črnem jedru na podganjem modelu za Parkinsonovo bolezen

Vesel, Jasmina (author), Glavan, Gordana (mentor), Pišler, Anja (co-mentor)

Cisteinska proteza katepsin X je pomemben dejavnik v degenerativnih procesih tekom normalnega staranja in v nevrodegenerativnih boleznih. Parkinsonova bolezen (PB) je pogosta nevrodegenerativna bolezen, za katero je značilen propad nigrostiatnih dopaminskih nevronov. V naši študiji smo raziskali vpliv nevrotoksina 6-hidroksidopamina (6-OHDA) na nivo izražanja katepsina X v črnem jedru in striatumu na podganjem modelu za PB. V nigrostriatno pot podganjih možganov smo unilateralno injicirali 6-OHDA in živali žrtvovali v različnih časovnih razmikih. Z metodo imunohistokemije smo dokazali povečano izražanje proteina katepsina X v črnem jedru 12, 24 in 48 ur po injiciranju 6-OHDA in s prenosom western dokazali povečan nivo izražanja proteina katepsina X v striatumu 12 ur po injiciranju 6-OHDA. Z dvojnim imunofluorescenčnim barvanjem smo 12 ur po injiciranju 6-OHDA dokazali katepsin X v dopaminskih celicah, 48 ur po injiciranju 6-OHDA pa ugotovili visok nivo katepsina X v lizosomih. Opazili smo tudi povečan nivo izražanja in aktivnosti encima katepsina X v črnem jedru 4 tedne po injiciranju 6-OHDA, pri čemer predpostavljamo, da gre za vnetni odziv povezan z nevrodegeneracijo. Naši rezultati se skladajo z domnevo, da ima katepsin X vlogo v patološkem procesu propadanja živčnih celic. Izsledki raziskave potrjujejo rezultate študije narejene na celičnih kulturah. S tem smo uspešno prenesli predhodne in vitro raziskave na in vivo model.

• Language: Slovenian
• Keywords: 6-OHDA, katepsin X, črno jedro, podganji model, Parkinsonova bolezen
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2017
• PID: 20.500.12556/RUL-99160
• COBISS.SI-ID: 4568143
• Publication date in RUL: 29.12.2017

Secondary language

• Language: English
• Title: Influence of 6-hydroxydopamine on cathepsin X level in substantia nigra on rat model of Parkinson's disease

Abstract

Cysteine protease cathepsin X is an important player in degenerative processes during normal aging and in neurodegenerative diseases. Parkinson's disease (PD) is a common neurodegenerative disease characterized by a degeneration of nigrostriatal dopaminergic neurons. In our study we investigated an influence of neurotoxin 6-hydroxydopamine (6-OHDA) on the level of protein cathepsin X in substantia nigra and striatum on a rat model of PD. We unilaterally injected 6-OHDA into nigrostriatal pathway of rat brain. Animals were later sacrificed at different times after the injection. With immunohistochemistry analysis we determined upregulation of protein level of cathepsin X in substantia nigra 12, 24 and 48 h after the injection of 6-OHDA and with Western blot analysis we determined upregulation of protein level of cathepsin X in striatum 12 h after the injection of 6-OHDA. With double immunofluorescence staining we confirmed localization of cathepsin X in dopaminergic neurons at 12 h and showed high level of cathepsin X in lysosomes at 48 h after treatment with 6-OHDA. We also noticed an upregulation of cathepsin X at 4 weeks for which we are assuming it is a consequence of neurodegeneration induced by inflammation. Our results are in accordance with cathepsin X role in pathological processes of neuronal degeneration. Results confirmed the results of study made on cell cultures. We confirmed the results obtained previously by in vitro studies also in vivo.

• Keywords: 6-OHDA, cathepsin X, substantia nigra, rat model, Parkinson's disease