The effect of selected cannabinoids on quantitative EEG, behaviour, and their therapeutic potential

Kanabinoidy a zejména CBD jsou dnes velmi diskutované téma s obrovským přesahem do mediálního prostoru. Jejich schopnost interagovat nejen kanabinoidními receptory, ale i dalšími drahami v výrazným "hypem", vede k testování kanabinoidů ve velmi širokém spektru indikací i přes nepříliš silnou evidenci účinků v klinické praxi. Dnes máme již více než 150 izolovaných kanabinoidů, ale výzkum je zaměřen pouze na několik hlavních kanabinoidů jako jsou například THC, CBD, kanabigerol, kanabichromen, hexahydrokanabinol. My jsme pro náš výzkum zvolili dva nejznámější kanabinoidy THC a CBD a naší klíčovou otázkou bylo, jako roli mají tyto kanabinoidy nejen samostatně, ale také jak spolu interagují při společném podání, jak ovlivňují chování a jak společně interagují závislosti na způsobu podání. Toto jsme souhrnně provedli v potkaních experimentech. Částečně překvapivě, jsme nepozorovali typickou kanabinoidní tetrádu (sníženou citlivost na bolestivé podněty, hypotermii, katalepsii a celkové snížení motorické aktivity) po podání THC, a proto jsme si položili otázku, co se stane, pokud místo THC použijeme potentnější a selektivnější syntetické kanabinoidy JWH 210. Dále v průběhu řešení disertační práce došlo k registraci CBD (Epidolex) pro léčbu epilepsie, konkrétně syndromu Dravetové a Lennox_Gaustova syndromu, a my...Cannabinoids and especially CBD are these days a highly discussed topic with a huge overlap in the media space. Their ability to interact not only with cannabinoid receptors, but also with other pathways, in combination with a huge "hype", leads, despite the not very strong evidence of their effects in clinical practice, to the testing of cannabinoids in a broad spectrum of indications. Nowadays, we have more than 150 isolated cannabinoids, but the research is focused only on the main cannabinoids like for example THC, CBD, cannabigerol, cannabichromen or hexahydrocannabinol. For our research, we have chosen the two most well- known cannabinoids THC and CBD, and our key question was not only what is the role of these cannabinoids individually, but, also, how do they interact when administered together, how do they influence behaviour and how do they interact in dependence on the method of administration. All these questions were addressed within our rat experiments. Somewhat surprisingly, we did not observe the typical cannabinoid tetrad (decreased sensitivity to painful stimuli, hypothermia, catalepsy, and overall decrease in motor activity) after the THC administration, so we wondered what would happen if we used more potent and selective synthetic cannabinoids JWH- 073, JWH-210, instead of THC....Katedra fyziologieDepartment of PhysiologyPřírodovědecká fakultaFaculty of Scienc

The effect of selected cannabinoids on quantitative EEG, behaviour, and their therapeutic potential

Cannabinoids and especially CBD are these days a highly discussed topic with a huge overlap in the media space. Their ability to interact not only with cannabinoid receptors, but also with other pathways, in combination with a huge "hype", leads, despite the not very strong evidence of their effects in clinical practice, to the testing of cannabinoids in a broad spectrum of indications. Nowadays, we have more than 150 isolated cannabinoids, but the research is focused only on the main cannabinoids like for example THC, CBD, cannabigerol, cannabichromen or hexahydrocannabinol. For our research, we have chosen the two most well- known cannabinoids THC and CBD, and our key question was not only what is the role of these cannabinoids individually, but, also, how do they interact when administered together, how do they influence behaviour and how do they interact in dependence on the method of administration. All these questions were addressed within our rat experiments. Somewhat surprisingly, we did not observe the typical cannabinoid tetrad (decreased sensitivity to painful stimuli, hypothermia, catalepsy, and overall decrease in motor activity) after the THC administration, so we wondered what would happen if we used more potent and selective synthetic cannabinoids JWH- 073, JWH-210, instead of THC....Kanabinoidy a zejména CBD jsou dnes velmi diskutované téma s obrovským přesahem do mediálního prostoru. Jejich schopnost interagovat nejen kanabinoidními receptory, ale i dalšími drahami v výrazným "hypem", vede k testování kanabinoidů ve velmi širokém spektru indikací i přes nepříliš silnou evidenci účinků v klinické praxi. Dnes máme již více než 150 izolovaných kanabinoidů, ale výzkum je zaměřen pouze na několik hlavních kanabinoidů jako jsou například THC, CBD, kanabigerol, kanabichromen, hexahydrokanabinol. My jsme pro náš výzkum zvolili dva nejznámější kanabinoidy THC a CBD a naší klíčovou otázkou bylo, jako roli mají tyto kanabinoidy nejen samostatně, ale také jak spolu interagují při společném podání, jak ovlivňují chování a jak společně interagují závislosti na způsobu podání. Toto jsme souhrnně provedli v potkaních experimentech. Částečně překvapivě, jsme nepozorovali typickou kanabinoidní tetrádu (sníženou citlivost na bolestivé podněty, hypotermii, katalepsii a celkové snížení motorické aktivity) po podání THC, a proto jsme si položili otázku, co se stane, pokud místo THC použijeme potentnější a selektivnější syntetické kanabinoidy JWH 210. Dále v průběhu řešení disertační práce došlo k registraci CBD (Epidolex) pro léčbu epilepsie, konkrétně syndromu Dravetové a Lennox_Gaustova syndromu, a my...Katedra fyziologieDepartment of PhysiologyFaculty of SciencePřírodovědecká fakult

The role of nitric oxide in CNS pathologies

Oxid dusnatý (NO) je významný mezibuněčný posel, široce využívaný v nervové soustavě. Moduluje velké množství fyziologických funkcí, účastní se imunitních odpovědí, ale zároveň při patologickém stavu může být odpovědný za široké množství škodlivých účinků. Cílem této práce je přiblížit formou literární rešerše široké téma oxidu dusnatého při patologii CNS. První část této práce přibližuje tvorbu NO, fyziologické funkce modulované NO, popisuje významnou signalizační dráhu NO-cGMP a zabývá se osudem produkovaného NO. Druhá část této práce je stručně zaměřena na metody výzkumu: použití a rozdělení inhibitorů NOS, geneticky modifikované myši, detekce NO a jeho produktů. Poslední část je věnována vybraným konkrétním patologickým stavům v CNS.Nitric oxide (NO) is significant intercellular messenger, widely used in nervous system. It is used to modulate large number of physiological functions and participates in immunity responses; however in pathological state it can be responsible for wide variety of harmful effects. The goal of my bachelor thesis is to illustrate the topic of nitric oxide in CNS pathology in the form of literature research. First part of this thesis shows the formation of NO, physiological functions influenced by NO, describes the main signaling pathway NO-cGMP and deals with the use of produced NO. Second part of this thesis briefly shows the research methods: the use and distribution of NOS inhibitors, genetically modified mice and detection of NO and its products. The last part is dedicated to selected pathological states in CNS.Katedra fyziologieDepartment of PhysiologyPřírodovědecká fakultaFaculty of Scienc

Behavioral and Pharmacokinetic Profile of Indole-Derived Synthetic Cannabinoids JWH-073 and JWH-210 as Compared to the Phytocannabinoid Δ9-THC in Rats

Synthetic cannabinoid compounds are marketed as “legal” marijuana substitutes, even though little is known about their behavioral effects in relation to their pharmacokinetic profiles. Therefore, in the present study we assessed the behavioral effects of systemic treatment with the two synthetic cannabinoids JWH-073 and JWH-210 and the phytocannabinoid Δ9-THC on locomotor activity, anxiety-like phenotype (in the open field) and sensorimotor gating (measured as prepulse inhibition of the acoustic startle response, PPI), in relation to cannabinoid serum levels. Wistar rats were injected subcutaneously (sc.) with JWH-073 (0.1, 0.5, or 5 mg/kg), JWH-210 (0.1, 0.5, or 5 mg/kg), Δ9-THC (1 or 3 mg/kg) or vehicle (oleum helanti) in a volume of 0.5 ml/kg and tested in the open field and PPI. Although JWH-073, JWH-210, Δ9-THC (and its metabolites) were confirmed in serum, effects on sensorimotor gating were absent, and locomotor activity was only partially affected. Δ9-THC (3 mg/kg) elicited an anxiolytic-like effect as suggested by the increased time spent in the center of the open field (p < 0.05). Our results further support the potential anxiolytic-like effect of pharmacological modulation of the endocannabinoid system

Activation of neuroglia after status epilepticus induced by intracerebroventricular application of 4-aminopyridine in the rat.

Epilepsy is a chronic disease characterized by spontaneous epileptic seizures. One percent of the world population is affected by the epilepsy. The existence of proper models is crucial for study of distinct types of epilepsy. We decided to deeply describe the model of status epilepticus (SE) induction in an adult rat by the intracerebroventricular application of 4-aminopyridine (4-AP). For description of this model we used the histology and immunohistochemistry methods, the western blot analysis focused on the microglial activation markers, and the series of behavioral tests to reveal the functional influence of SE in chronical experiment (8 months). We described the ability of the 4-AP to induce SE in this model. After SE, neuron degeneration and microglial activation appeared. The areas of degenerated neurons strongly corresponded to the regions with activated microglias. By the western blotting we demonstrated the oxidative stress by an increase of 3-nitrotyrosine already 2 hours after SE. Microglial activation detected by the immunohistochemistry correlated to the observed increase in IL-1β and CD68 production after SE. Considering these results we verified the functional influence of the SE in chronical experiment. We did not observed any significant changes of cognitive and motoric functions..

Activation of neuroglia after status epilepticus induced by intracerebroventricular application of 4-aminopyridine in the rat.

Epilepsy is a chronic disease characterized by spontaneous epileptic seizures. One percent of the world population is affected by the epilepsy. The existence of proper models is crucial for study of distinct types of epilepsy. We decided to deeply describe the model of status epilepticus (SE) induction in an adult rat by the intracerebroventricular application of 4-aminopyridine (4-AP). For description of this model we used the histology and immunohistochemistry methods, the western blot analysis focused on the microglial activation markers, and the series of behavioral tests to reveal the functional influence of SE in chronical experiment (8 months). We described the ability of the 4-AP to induce SE in this model. After SE, neuron degeneration and microglial activation appeared. The areas of degenerated neurons strongly corresponded to the regions with activated microglias. By the western blotting we demonstrated the oxidative stress by an increase of 3-nitrotyrosine already 2 hours after SE. Microglial activation detected by the immunohistochemistry correlated to the observed increase in IL-1β and CD68 production after SE. Considering these results we verified the functional influence of the SE in chronical experiment. We did not observed any significant changes of cognitive and motoric functions..

The effect of selected cannabinoids on quantitative EEG, behaviour, and their therapeutic potential

Cannabinoids and especially CBD are these days a highly discussed topic with a huge overlap in the media space. Their ability to interact not only with cannabinoid receptors, but also with other pathways, in combination with a huge "hype", leads, despite the not very strong evidence of their effects in clinical practice, to the testing of cannabinoids in a broad spectrum of indications. Nowadays, we have more than 150 isolated cannabinoids, but the research is focused only on the main cannabinoids like for example THC, CBD, cannabigerol, cannabichromen or hexahydrocannabinol. For our research, we have chosen the two most well- known cannabinoids THC and CBD, and our key question was not only what is the role of these cannabinoids individually, but, also, how do they interact when administered together, how do they influence behaviour and how do they interact in dependence on the method of administration. All these questions were addressed within our rat experiments. Somewhat surprisingly, we did not observe the typical cannabinoid tetrad (decreased sensitivity to painful stimuli, hypothermia, catalepsy, and overall decrease in motor activity) after the THC administration, so we wondered what would happen if we used more potent and selective synthetic cannabinoids JWH- 073, JWH-210, instead of THC...

Anticonvulsive Effects and Pharmacokinetic Profile of Cannabidiol (CBD) in the Pentylenetetrazol (PTZ) or N-Methyl-D-Aspartate (NMDA) Models of Seizures in Infantile Rats

In spite of use of cannabidiol (CBD), a non-psychoactive cannabinoid, in pediatric patients with epilepsy, preclinical studies on its effects in immature animals are very limited. In the present study we investigated anti-seizure activity of CBD (10 and 60 mg/kg administered intraperitoneally) in two models of chemically induced seizures in infantile (12-days old) rats. Seizures were induced either with pentylenetetrazol (PTZ) or N-methyl-D-aspartate (NMDA). In parallel, brain and plasma levels of CBD and possible motor adverse effects were assessed in the righting reflex and the bar holding tests. CBD was ineffective against NMDA-induced seizures, but in a dose 60 mg/kg abolished the tonic phase of PTZ-induced generalized seizures. Plasma and brain levels of CBD were determined up to 24 h after administration. Peak CBD levels in the brain (996 ± 128 and 5689 ± 150 ng/g after the 10- and 60-mg/kg doses, respectively) were reached 1–2 h after administration and were still detectable 24 h later (120 ± 12 and 904 ± 63 ng/g, respectively). None of the doses negatively affected motor performance within 1 h after administration, but CBD in both doses blocked improvement in the bar holding test with repeated exposure to this task. Taken together, anti-seizure activity of CBD in infantile animals is dose and model dependent, and at therapeutic doses CBD does not cause motor impairment. The potential risk of CBD for motor learning seen in repeated motor tests has to be further examined

Adenosine Kinase Isoforms in the Developing Rat Hippocampus after LiCl/Pilocarpine Status Epilepticus

LiCl/pilocarpine status epilepticus (SE) induced in immature rats leads, after a latent period, to hippocampal hyperexcitability. The excitability may be influenced by adenosine, which exhibits anticonvulsant activity. The concentration of adenosine is regulated by adenosine kinase (ADK) present in two isoforms—ADK-L and ADK-S. The main goal of the study is to elucidate the changes in ADK isoform expression after LiCl/pilocarpine SE and whether potential changes, as well as inhibition of ADK by 5-iodotubercidin (5-ITU), may contribute to changes in hippocampal excitability during brain development. LiCl/pilocarpine SE was elicited in 12-day-old rats. Hippocampal excitability in immature rats was studied by the model of hippocampal afterdischarges (ADs), in which we demonstrated the potential inhibitory effect of 5-ITU. ADs demonstrated significantly decreased hippocampal excitability 3 days after SE induction, whereas significant hyperexcitability after 20 days compared to controls was shown. 5-ITU administration showed its inhibitory effect on the ADs in 32-day-old SE rats compared to SE rats without 5-ITU. Moreover, both ADK isoforms were examined in the immature rat hippocampus. The ADK-L isoform demonstrated significantly decreased expression in 12-day-old SE rats compared to the appropriate naïve rats, whereas increased ADK-S isoform expression was revealed. A decreasing ADK-L/-S ratio showed the declining dominance of ADK-L isoform during early brain development. LiCl/pilocarpine SE increased the excitability of the hippocampus 20 days after SE induction. The ADK inhibitor 5-ITU exhibited anticonvulsant activity at the same age. Age-related differences in hippocampal excitability after SE might correspond to the development of ADK isoform levels in the hippocampus