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Amaryllidaceae alkaloids of haemanthamine structural type and their semisynthetic derivatives as potential drugs in the treatment of Alzheimer's disease.
Peřinová, Rozálie ; Cahlíková, Lucie (advisor) ; Lapčík, Oldřich (referee) ; Mučaji, Pavel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany Candidate: Ing. Rozálie Peřinová Supervisor: prof. Ing. Lucie Cahlíková, Ph.D. Title of doctoral thesis: Amaryllidaceae alkaloids of haemanthamine structural type and their semisynthetic derivatives as potential drugs in the treatment of Alzheimer's disease. To deepen the knowledge about the Amaryllidaceae alkaloid haemanthamine, which was isolated at our workplace as part of previous phytochemical studies, derivatives of this alkaloid were synthesized. First, series of aliphatic (3-12) and aromatic ester derivatives (13-66) were prepared, and then, to compare the structure-activity relationship, ether derivatives (67-80) were prepared from the most active substituents. All synthesized compounds were identified using the following structural analysis methods: NMR, HPLC/MS, and HRMS, including testing physical properties such as optical rotatability. After structure confirmation, all derivatives were subjected to screening studies for their inhibitory potential against hAChE and hBuChE. The selected derivatives were tested for their inhibitory potential against another enzyme, GSK-3β, which plays a significant role in the pathogenesis of AD. In cooperation with the Faculty of Medicine in...
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Studies on the molecular mechanisms of cardioprotective effects of morphine
Škrabalová, Jitka ; Novotný, Jiří (advisor) ; Nováková, Olga (referee) ; Hlaváčková, Markéta (referee)
Acute and chronic morphine administration can significantly reduce ischemia- reperfusion injury of the rat heart. However, the molecular mechanisms mediating the protective effect of morphine are not yet fully elucidated. Concurrently, there is a lack of information about the effects of the long-term action of morphine on heart tissue. Therefore, in the first part of the project, we studied the effect of long-term administration of high doses of morphine (10 mg/kg/day, 10 days) on rat heart tissue. In the second part of the project, we investigated the effect of 1 mM morphine on viability and redox state of rat cardiomyoblast cell line H9c2 that was influenced by oxidative stress elicited by exposure to 300 μM tert-butyl hydroperoxide (t-BHP). Our experiments have shown that long-term morphine administration affected neither the amount nor the affinity of myocardial β-adrenergic receptors (β-AR), but almost doubled the number of the dominant isoforms of myocardial adenylyl cyclase (AC) V/VI and led to supersensitization of AC. At the same time, proteomic analyses revealed that long-term morphine administration was associated with significant changes in the left ventricular proteome. In particular, there was an increase in the expression of heat shock proteins (HSP). Increased expression of HSP27...
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Alzheimer's disease-related changes in phosphorylation of collapsin response mediator protein 2 (CRMP2) and their effect on neuronal function
Škrlantová, Alice ; Balaštík, Martin (advisor) ; Balík, Aleš (referee)
CRMP2 was first identified in 1995 as a mediator of Sema3A signalization pathway which leads to axon growth cone collapse. Since then CRMP2 was designated as an essential cue during neuronal polarity estabilishment and neuronal growth in embryonic life. CRMP2 was also found hyperphosphorylated in NFT's and this finding led to further research of CRMP2 function in the pathogenesis of AD. The activity and proper function of CRMP2 is regulated by phosphorylation and a deeper look into the mechanism of this modification is necessary for understanding how CRMP2 influences the function of neural cells. In this thesis I focus on signaling pathways, kinases and interaction partners of CRMP2 and describe how aberrant regulation of these interactions leads to Alzheimerʼs disease development.
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Studies on the molecular mechanisms of cardioprotective effects of morphine
Škrabalová, Jitka
Acute and chronic morphine administration can significantly reduce ischemia- reperfusion injury of the rat heart. However, the molecular mechanisms mediating the protective effect of morphine are not yet fully elucidated. Concurrently, there is a lack of information about the effects of the long-term action of morphine on heart tissue. Therefore, in the first part of the project, we studied the effect of long-term administration of high doses of morphine (10 mg/kg/day, 10 days) on rat heart tissue. In the second part of the project, we investigated the effect of 1 mM morphine on viability and redox state of rat cardiomyoblast cell line H9c2 that was influenced by oxidative stress elicited by exposure to 300 μM tert-butyl hydroperoxide (t-BHP). Our experiments have shown that long-term morphine administration affected neither the amount nor the affinity of myocardial β-adrenergic receptors (β-AR), but almost doubled the number of the dominant isoforms of myocardial adenylyl cyclase (AC) V/VI and led to supersensitization of AC. At the same time, proteomic analyses revealed that long-term morphine administration was associated with significant changes in the left ventricular proteome. In particular, there was an increase in the expression of heat shock proteins (HSP). Increased expression of HSP27...
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Alkaloids of the Amaryllidaceae family as potential drugs in therapy of diseases of affluence
Breiterová, Kateřina ; Cahlíková, Lucie (advisor) ; Lapčík, Oldřich (referee) ; Nagy, Milan (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Mgr. Kateřina Breiterová Supervisor: Assoc. prof. Ing. Lucie Cahlíková PhD. Title of Doctoral Thesis: Alkaloids of the Amaryllidaceae family as potential drugs in therapy of diseases of affluence Key words: alkaloids, Amaryllidaceae, analogues, AChE, BuChE, POP, GSK-3β, cell cycle progression, apoptosis Narcissus cv. Professor Einstein was chosen based on results of previous screening studies for detailed phytochemical work for the purpose of isolation of the widest range of AmA. From 34,3 kg of fresh bulbs was obtained 31,7 g of purified alkaloidal extract, which was processed using column chromatography with stepwise elution by light petrol, chloroform and ethanol in different ratios to almost 500 fractions. These fractions were fused into 27 subfractions, which were processed by preparative TLC, vacuum column chromatography and crystallization. Finally, 25 pure alkaloids were isolated. All compounds were identified by GC-MS, ESI-MS, NMR, optical rotation and literature. One compound was identified as a new unpublished alkaloid of lycorine structure type. All alkaloids isolated in sufficient amount were tested for their biological activities associated with Alzheimer's disease (inhibition of...
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Studies on the molecular mechanisms of cardioprotective effects of morphine
Škrabalová, Jitka
Acute and chronic morphine administration can significantly reduce ischemia- reperfusion injury of the rat heart. However, the molecular mechanisms mediating the protective effect of morphine are not yet fully elucidated. Concurrently, there is a lack of information about the effects of the long-term action of morphine on heart tissue. Therefore, in the first part of the project, we studied the effect of long-term administration of high doses of morphine (10 mg/kg/day, 10 days) on rat heart tissue. In the second part of the project, we investigated the effect of 1 mM morphine on viability and redox state of rat cardiomyoblast cell line H9c2 that was influenced by oxidative stress elicited by exposure to 300 μM tert-butyl hydroperoxide (t-BHP). Our experiments have shown that long-term morphine administration affected neither the amount nor the affinity of myocardial β-adrenergic receptors (β-AR), but almost doubled the number of the dominant isoforms of myocardial adenylyl cyclase (AC) V/VI and led to supersensitization of AC. At the same time, proteomic analyses revealed that long-term morphine administration was associated with significant changes in the left ventricular proteome. In particular, there was an increase in the expression of heat shock proteins (HSP). Increased expression of HSP27...
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Studies on the molecular mechanisms of cardioprotective effects of morphine
Škrabalová, Jitka ; Novotný, Jiří (advisor) ; Nováková, Olga (referee) ; Hlaváčková, Markéta (referee)
Acute and chronic morphine administration can significantly reduce ischemia- reperfusion injury of the rat heart. However, the molecular mechanisms mediating the protective effect of morphine are not yet fully elucidated. Concurrently, there is a lack of information about the effects of the long-term action of morphine on heart tissue. Therefore, in the first part of the project, we studied the effect of long-term administration of high doses of morphine (10 mg/kg/day, 10 days) on rat heart tissue. In the second part of the project, we investigated the effect of 1 mM morphine on viability and redox state of rat cardiomyoblast cell line H9c2 that was influenced by oxidative stress elicited by exposure to 300 μM tert-butyl hydroperoxide (t-BHP). Our experiments have shown that long-term morphine administration affected neither the amount nor the affinity of myocardial β-adrenergic receptors (β-AR), but almost doubled the number of the dominant isoforms of myocardial adenylyl cyclase (AC) V/VI and led to supersensitization of AC. At the same time, proteomic analyses revealed that long-term morphine administration was associated with significant changes in the left ventricular proteome. In particular, there was an increase in the expression of heat shock proteins (HSP). Increased expression of HSP27...
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Alzheimer's disease-related changes in phosphorylation of collapsin response mediator protein 2 (CRMP2) and their effect on neuronal function
Škrlantová, Alice ; Balaštík, Martin (advisor) ; Balík, Aleš (referee)
CRMP2 was first identified in 1995 as a mediator of Sema3A signalization pathway which leads to axon growth cone collapse. Since then CRMP2 was designated as an essential cue during neuronal polarity estabilishment and neuronal growth in embryonic life. CRMP2 was also found hyperphosphorylated in NFT's and this finding led to further research of CRMP2 function in the pathogenesis of AD. The activity and proper function of CRMP2 is regulated by phosphorylation and a deeper look into the mechanism of this modification is necessary for understanding how CRMP2 influences the function of neural cells. In this thesis I focus on signaling pathways, kinases and interaction partners of CRMP2 and describe how aberrant regulation of these interactions leads to Alzheimerʼs disease development.
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Circadian regulation of STAT3 protein in the SCN and it's activation by leptin in the SCN, other parts of hypothalamus and the pineal gland
Moníková, Veronika ; Bendová, Zdeňka (advisor) ; Jelínková, Dana (referee)
JAK/STAT signaling pathway is one of the most studied intracellular cascades transmitting signals from the extracellular environment to the cell nucleus in order to affect expression of target genes. Circadian clocks localized in the suprachiasmatic nuclei (SCN) of the hypothalamus are sensitive especially to light but they can respond to non-photic stimuli such as growth factors, opioids, leptin and cytokines that have been demonstrated to perform its function via the JAK/STAT signaling pathway. The recent findings of our laboratory demonstrated that STAT3 protein is highly produced by SCN of rat. Primary aim of our experiments was to test the circadian regulation of STAT3 production in SCN and describe the effect of exogenously administered leptin on STAT3 phosphorylation in the SCN, pineal gland and hypothalamic structures responsible for regulated feeding behavior and energy metabolism. Because activation of leptin receptors may stimulate a number of other signaling cascades, we chose phosphorylated forms of kinase ERK1/2 and GSK-3β as other markers of intracellular changes after administration of leptin in the studied structures. Our results proved rhythmic production of STAT3 protein in SCN of rat and indicated circadian regulation of sensitivity to leptin in hypothalamic structures. The data...
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