Djelovanje gabapentina i alkohola na staničnu kulturu HEK 293

master's thesis

• Title (english): The effects of gabapentin and alcohol on HEK 293 cell culture
• Author: Marina Morić
• Mentor: Dubravka Švob Štrac (mentor)
• Mentor: Dubravka Hranilović (sumentor)
• Committee member: Dubravka Hranilović (predsjednik povjerenstva)
• Committee member: Dubravka Švob Štrac (član povjerenstva)
• Committee member: Jasna Hrenović (član povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2014-05-16, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Antikonvulzivni lijek gabapentin pokazuje pozitivne učinke u liječenju alkoholizma i njegovih posljedica. Iako mehanizmi odgovorni za različite učinke gabapentina nisu potpuno jasni, istraživanja upućuju da bi neuroprotektivno djelovanje gabapentin mogao ostvarivati putem receptora GABAA. Tretman etanolom kroz 96 sati u koncentracijama višim od 100 mM značajno smanjuje broj stanica HEK 293, netransfeciranih i transfeciranih receptorima α1β2γ2S GABAA. Citotoksični učinak etanola poništen je istovremenom primjenom 1 μM gabapentina kod transfeciranih, ali ne i netransfeciranih stanica HEK 293. Primjena 100 mM etanola kroz 96 sati izazvala je značajan porast broja veznih mjesta za konvulzive i benzodiazepine rekombinantnih receptora GABAA, dok se primjenom etanola u kombinaciji s 1 μM gabapentinom, 100 μM bikukulinom ili njihovom kombinacijom, broj receptorskih veznih mjesta vratio na kontrolne razine. Primjena etanola, kao i bikukulina izazvala je alosteričko razdvajanje veznih mjesta za GABA-u i benzodiazepine, dok je istovremena primjena gabapentina spriječila etanolom i bikukulinom izazvano slabljenje receptorskih funkcionalnih veza. Ekspresija glasničke RNA podjedinica α1, β2 i γ2S receptora GABAA nije se značajno promijenila pod dugotrajnim utjecajem etanola ili gabapentina, već samo njihove kombinacije. Iako rezultati podržavaju pretpostavku o uključenosti receptora GABAA u djelovanju gabapentina, potrebna su daljnja istraživanja za utvrđivanje mehanizma protektivnog učinka gabapentina u slučaju citotoksičnog djelovanja etanola.

Abstract (english)

The anticonvulsant drug gabapentin showed positive results in the treatment of alcoholism and its consequences. Although mechanisms of gabapentin action are not fully understood, studies suggested that gabapentin neuroprotective effects could be achieved via GABAA receptors. Treatment with ethanol at concentrations higher than 100 mM for 96 hours reduced the number of HEK 293 cells, untransfected or transfected with α1β2γ2S GABAA receptors. The cytotoxic effect of ethanol was counteracted by co-administration of 1 μM gabapentin in transfected, but not in untransfected cells. Exposure to 100 mM ethanol induced an increase in the number of binding sites for benzodiazepines and convulsants, while this effect was reversed by simultaneous exposure to 1 μM gabapentin, 100 μM bicuculline or their combination. Administration of ethanol, as well as bicuculline, induced functional allosteric uncoupling of GABA and benzodiazepine binding site, which was prevented by simultaneous gabapentin treatment. Increase in mRNA expression of α1, β2 and γ2S GABAA receptor subunits was not influenced by the exposure to ethanol or gabapentin, but only by their co-administration. Although results support the hypothesis of the involvement of GABAA receptors in the actions of gabapentin, further research is needed to elucidate the mechanisms of gabapentin protective effects against ethanol-induced cytotoxicity.

• Keywords: gabapentin; dugotrajna primjena; alkohol; kultura Stanica HEK 293; rekombinantni receptori GABAA; citotoksičnost
• Keywords (english): gabapentin; long-term treatment; alcohol; recombinant GABAA receptors; HEK 293 cell culture; cytotoxicity
• Language: croatian
• URN:NBN: urn:nbn:hr:217:801174
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-26 10:45:08