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Opioid-induced microbial dysbiosis
Popjak, Šimon ; Drastichová, Zdeňka (advisor) ; Stehlíková, Zuzana (referee)
The aim of this bachelor thesis is to describe the consequences of opioids in relation to the gut microbiome. Chronic opioid use induces microbial dysbiosis in the gut, where both functional and structural changes in the composition of the microbiome occur. This disrupts gut homeostasis and leads to diseases. The diversity of microbial organisms decreases and the expression of neurotransmitters changes. Opioids interfere with the microbiomeň-gut-brain axis, which normally functions in symbiosis. They alter the composition of the gut microbiota, affect the function and structure of the gut epithelium and the expression of neurotransmitters in the nervous system. This thesis focuses primarily on the action of morphine, the action of which has been desribed the most. Keywords: opioids, microbiome, dysbiosis, microbiome-gut-brain axis, gut homeostasis
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The effect of morphine withdrawal on the catecholaminergic and serotonergic neurotransmitter system in rat brain
Nováková, Daniela ; Novotný, Jiří (advisor) ; Drastichová, Zdeňka (referee)
The aim of this diploma thesis is to study the effect of morphine withdrawal on catecholaminergic and serotonergic neurotransmitter system in rat brain. Theoretical part of this thesis summarizes basic information known about principles of neurotransmission with focus on the catecholaminergic and serotonergic system, metabolism of its components, their signaling, relevant receptors, their distribution, and especially their effect on morphine dependence and subsequent withdrawal. It also summarizes briefly principles of opioid signaling and outlines the findings yet known about neurochemical analysis of the transmitter systems mentioned above. Experimental part of this thesis is focused on the optimization of the method of high performance liquid chromatography with fluorescence detection and its subsequent use to determine basic components of catecholaminergic and serotonergic neurotransmitter system in samples of different part of brain of rats affected by intraperitoneal administration of morphine sulphate, and its subsequent withdrawal and correspondings unaffected control rats. The expression of selected catecholaminergic receptors in identical samples is also detected. This thesis succesfully implements gradient into the originally isocratic method of high performance liquid chromatography...
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Cell signalling and molecular complexes of the TRH receptor
Drastichová, Zdeňka
The first part of this thesis is preoccupied with the identification of protein alterations in the membrane fraction of HEK293-E2M11 cells after prolonged TRH treatment. The isolated membrane fraction enriched in plasma membranes contained high amounts of Na+ ,K+ -ATPase, TRH receptor and G-proteins compared to the postnuclear supernatant. By using 2D electrophoresis and mass spectrometry, the levels of 42 proteins were identified to be altered in samples of PM-enriched fractions from TRH-treated (16 h; 10 μM) cells. Out of these proteins only ezrin and stomatin-like 2 are known to be localized in the plasma membrane. Five proteins (mitofilin, MTHSP75, prohibitin, stomatin like-2, peroxiredoxin III) whose levels were increased after the prolonged TRH treatment represent proteins localized in mitochondria. All of them are important for proper structure and function of mitochondria. The ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax was markedly higher in cells treated with TRH compared to control cells. Hence, it can be concluded that prolonged TRH treatment may significantly affect mitochondrial membrane and function of mitochondria. The second part of this thesis deals with the identification of molecular protein complexes of TRH-R and/or Gq/11 protein. The presumed effects of TRH on the...
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Cell signalling and molecular complexes of the TRH receptor
Drastichová, Zdeňka
The first part of this thesis is preoccupied with the identification of protein alterations in the membrane fraction of HEK293-E2M11 cells after prolonged TRH treatment. The isolated membrane fraction enriched in plasma membranes contained high amounts of Na+ ,K+ -ATPase, TRH receptor and G-proteins compared to the postnuclear supernatant. By using 2D electrophoresis and mass spectrometry, the levels of 42 proteins were identified to be altered in samples of PM-enriched fractions from TRH-treated (16 h; 10 μM) cells. Out of these proteins only ezrin and stomatin-like 2 are known to be localized in the plasma membrane. Five proteins (mitofilin, MTHSP75, prohibitin, stomatin like-2, peroxiredoxin III) whose levels were increased after the prolonged TRH treatment represent proteins localized in mitochondria. All of them are important for proper structure and function of mitochondria. The ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax was markedly higher in cells treated with TRH compared to control cells. Hence, it can be concluded that prolonged TRH treatment may significantly affect mitochondrial membrane and function of mitochondria. The second part of this thesis deals with the identification of molecular protein complexes of TRH-R and/or Gq/11 protein. The presumed effects of TRH on the...
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Cell signalling and molecular complexes of the TRH receptor
Drastichová, Zdeňka ; Novotný, Jiří (advisor) ; Hodný, Zdeněk (referee) ; Říčný, Jan (referee)
1 Summary The first part of this thesis is preoccupied with the identification of protein alterations in the membrane fraction of HEK293-E2M11 cells after prolonged TRH treatment. The isolated membrane fraction enriched in plasma membranes contained markedly increased the amount of Na,K-ATPase, TRH receptor and G-proteins compared to the postnuclear supernatant. By using 2D electrophoresis and mass spectrometry, the levels of 42 proteins were identified to be altered in samples of PM- enriched fractions from TRH-treated (16 h; 10 μM) cells. Out of these proteins only ezrin and stomatin-like 2 are known to be localized in the plasma membrane. Five proteins (mitofilin, MTHSP75, prohibitin, stomatin like-2, peroxiredoxin III) whose levels were increased after the prolonged TRH treatment represent proteins localized in mitochondria. All of them are important for proper structure and function of mitochondria. The ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax was markedly higher in cells treated with TRH than in control untreated cells. Hence, it can be concluded that prolonged TRH treatment may significantly affect mitochondrial membrane and function of mitochondria. The second part of this thesis deals with the identification of molecular protein complexes of TRH-R and/or Gq/11 protein. The presumed...
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