Neuroprotective mechanisms of nevirapine and efavirenz in a model of neurodegeneration
- Authors: Zheve, Georgina Teurai
- Date: 2008
- Subjects: HIV infections -- Treatment AIDS (Disease) -- Treatment AIDS dementia complex -- Treatment Nervous system -- Degeneration -- Treatment Melatonin Neurotoxic agents Quinolinic acid
- Language: English
- Type: Thesis , Masters , MPharm
- Identifier: vital:3807 , http://hdl.handle.net/10962/d1003285
- Description: AIDS Dementia Complex (ADC) is a neurodegenerative disorder implicated in HIV-1 infection that is associated with elevated levels of the neurotoxin, quinolinic acid (QA) which causes a cascade of events to occur, leading to the production of reactive oxygen species (ROS), these being ultimately responsible for oxidative neurotoxicity. In clinical studies, Non-nucleoside reverse transcriptase inhibitors (NNRTIs), efavirenz (EFV) and nevirapine (NVP) have been shown to potentially delay the progressive degeneration of neurons, thus reducing the frequency and neurological deficits associated with ADC. Despite these neuroprotective implications, there is still no biochemical data to demonstrate the mechanisms through which these agents offer neuroprotection. The present study aims to elucidate and further characterize the possible antioxidant and neuroprotective mechanisms of NVP and EFV in vitro and in vivo, using QA-induced neurotoxicity as a model. Research has demonstrated that antioxidants and metal chelators have the ability to offer neuroprotection against free radical induced injury and may be beneficial in the prevention or treatment of neurodegeneration. Hence the antioxidant and metal binding properties of these agents were investigated respectively. Inorganic studies, including the 1, 1-diphenyl-2 picrylhydrazyl (DPPH) assay, show that these agents readily scavenge free radicals in vitro, thus postulating the antioxidant property of these agents. The enhancement of superoxide radical generation and iron mediated Fenton reaction by QA is related to lipid peroxidation in biological systems, the extent of which was assayed using the nitroblue tetrazolium and thiobarbituric acid method respectively. Both agents significantly curtail QA-induced lipid peroxidation and potentially scavenge superoxide anions generated by cyanide in vitro. Furthermore, in vivo results demonstrate the ability of NVP and EFV to protect hippocampal neurons against lipid peroxidation induced by QA and superoxide radicals generated as a consequence thereof. The alleviation of QA-induced oxidative stress in vitro possibly occurs through the binding of iron (II) and / or iron (III), and this argument is further strengthened by the ability of EFV and not NVP to reduce iron (II)-induced lipid peroxidation in vitro directly. In addition the ferrozine and electrochemistry assay were used to measure the extent of iron (II) Fe[superscript 2+] and iron (III) Fe[superscript 3+] chelation activity. Both assays demonstrate that these agents bind iron (II) and iron (III), and prevent redox recycling of iron and subsequent complexation of Fe[superscript 2+] with QA which enhances neuronal damage. Both NNRTIs inhibit the endogenous biosynthesis of QA by inhibiting liver tryptophan 2, 3-dioxygenase activity in vivo and subsequently increasing hippocampal serotonin levels. Furthermore, these agents reduce the turnover of hippocampal serotonin to 5-hydroxyindole acetic acid. NVP and not EFV increase 5-hydroxyindole acetic acid and norepinephrine levels in the hippocampus. The results of the pineal indole metabolism study show that NVP increases the synthesis of melatonin, but decreases N-acetylserotonin, 5-hydroxyindole acetic acid and 5-hydroxytryptophol levels. Furthermore, it shows that EFV decreases 5-hydroxyindole acetic acid and melatonin synthesis. Behavioural studies using a Morris water maze show that the post-treatment of rats with NVP and EFV significantly improves QA-induced spatial memory deficits in the hippocampus. This study therefore provides novel information regarding the neuroprotective mechanisms of NVP and EFV. These findings strengthen the argument that these NNRTIs not only have antiviral effects but possess potential neuroprotective properties, which may contribute to the effectiveness of these drugs in the treatment of ADC.
- Full Text:
- Date Issued: 2008
An investigation into the antioxidative potential and regulatory aspects of liver tryptophan 2,3-dioxygenase by tryptophan and related analogues
- Authors: Antunes, Ana Paula Martins
- Date: 1998
- Subjects: Tryptophan -- Physiological effect , Antioxidants , Liver
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4010 , http://hdl.handle.net/10962/d1004070 , Tryptophan -- Physiological effect , Antioxidants , Liver
- Description: The amino acid, tryptophan, obtained through dietary means, is metabolised by the enzymes tryptophan 2,3-dioxygenase (TDO), indoleamine 2,3-dioxygenase (IDO) and tryptophan hydroxylase. All the enzymes have an effect on circulating tryptophan levels, especially TDO, since it is the major site of tryptophan catabolism in the liver and results in the production of kynurenine metabolites, viz. kynurenine, kynurenic acid, 3-hydroxyanthranilic acid and quinolinic acid. Extrahepatically, IDO is responsible for the synthesis of the kynurenine metabolites. Tryptophan 2,3-dioxygenase and IDO activity is increased by hormones or substrates such as tryptophan, and inflammation, in the case of IDO. Tryptophan availability for serotonin (5-HT) synthesis by the enzyme tryptophan hydroxylase is primarily dependent on TDO activity. A study was attempted in order to ascertain whether any of the endogenous metabolites of the kynurenine and serotonergic pathways would be able to inhibit TDO activity. Results showed that although the kynurenines had no effect, the indoleamines, except for the indoleacetic acids, were able to reduce TDO activity. 6-Methoxy-2-benzoxazolinone (6-MBOA), a structural analogue to melatonin, was the most potent inhibitor with a reduction in activity of 55 % compared with the control. The pineal gland in the rat brain has been shown to have the highest IDO activity. With induction, the kynurenine metabolite concentrations of kynurenic acid and quinolinic acid are increased. The effects of both compounds were determined on the serotonergic pathway. Although kynurenic acid produced no significant effect, quinolinic acid significantly reduced N-acetylserotonin and melatonin synthesis at concentrations of lOJLM and 100 JLM respectively. Many authors have implicated oxygen derived species as causative agents in the important neurodegenerative disorders such as Parkinson's and Huntington's disease. Increased radical generation and lipid peroxidation have been suggested to be responsible for the toxic destruction of neurons, especially in the brain because of its high lipid content and oxygen demand. The brain is therefore vulnerable to oxidative attack. During inflammatory diseases, IDO is induced with a resultant increase in kynurenines. This study was also an attempt at determining the effect of kynurenines on lipid peroxidation. All metabolites of the kynurenine pathway were able to induce lipid peroxidation significantly. The antioxidative potential of various tryptophan analogues, viz. serotonin, melatonin and 6-methoxy-2-benzoxazolinone, was determined using quinolinic acid-induced lipid peroxidation. Serotonin, melatonin and 6-MBOA were able to significantly reduce quinolinic acid-induced lipid peroxidation.
- Full Text:
- Date Issued: 1998
The pineal gland as a model to elucidate the primary mode of action of sympathoactive agents
- Authors: Welman, Alan David
- Date: 1991
- Subjects: Pineal gland , Cythochemistry
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3876 , http://hdl.handle.net/10962/d1001610
- Description: An attempt was made to use the pineal gland as a model for the study of the primary mode of action of sympathoactive agents. Two drugs were investigated, viz. alpha-methyldopa and ephedrine whose mode of action is not entirely clear. Organ cultures of pineal glands from rats treated chronically with alpha-methyldopa showed enhanced conversion of radioactive serotonin to melatonin (aMT) , as well as its precursor Nacetylserotonin (aHT). This treatment was also found to raise Nacetyltransferase (NAT) activity. These increases associated with alpha-methyldopa treatment were further enhanced by the beta-adrenergic agonist, isoproterenol, suggesting a supersensitivity-type effect occurring at the level of the beta-receptor. A subsequent binding study, however, showed a decrease in beta-receptor binding with exposure to alpha-methyldopa, providing mitigating evidence against the occurrence of a supersensitivity phenomenon. It is possible that a metabolite of alpha-methyldopa acts as an alpha 1 and beta-adrenergic agonist, resulting in greater melatonin (aMT) and N-acetylserotonin (aHT) synthesis than by a beta-adrenergic agonist, isoproterenol. Combined treatment of pineals with alpha-methyldopa and an alphareceptor blocker, phentolamine, resulted in melatonin (aMT) , Nacetylserotonin (aHT) , and N-acetyltransferase (NAT) activity levels which were lower than those obtained with alpha-methyldopa treatment alone, thus confirming the alpha-adrenergic activity of the metabolite of alpha-methyldopa. Additional pineal metabolites were isolated and measured simultaneously in the organ culture experiments. Organ cultures of rat pineal glands treated with ephedrine showed raised levels of melatonin (aMT) and N-acetylserotonin (aHT). Treatment with ephedrine also produced raised N-acetyltransferase activity. A further enhancement of these parameters was induced by norepinephrine, suggesting a supersensitivity-type effect occurring at the level of the beta-adrenergic receptor. Rats were treated with reserpine (a norepinephrine depleter) and the pineals exposed to ephedrine. Endogenous norepinephrine normally released by the action of ephedrine was thus absent, and under these conditions, levels of melatonin (aMT) and N-acetylserotonin (aHT) were reduced. N-acetyltransferase (NAT) activity was also reduced, but maintained levels pointing to substantial adrenergic activity of ephedrine as well as norepinephrine released by virtue of the drug's action. A subsequent binding study showed a decrease in beta-adrenergic receptor binding with exposure to ephedrine and a further decrease in ephedrine treated pineals from reserpine treated rats, thus ruling out the occurrence of a supersensitivity phenomenon. It is possible that both ephedrine and released norepinephrine have alpha- and beta-receptor activity. Additional pineal metabolites were isolated and measured in the organ culture experiments. A 16-hour time profile of the production of melatonin (aMT) and N-acetylserotonin (aHT) with norepinephrine and ephedrine treatment provided useful information regarding the course of action of the two agents. A pineal cell-culture system was developed and exposed to ephedrine and norepinephrine. N-acetyltransferase (NAT) activity levels measured after exposure to these agents were raised, confirming the adrenergic activity of both in the model. Finally, an HPLC system coupled to a UV detector was used in an attempt to measure melatonin (aMT) extracted from pineal organ culture media. The results showed that melatonin could be measured by this method, however, a more sensitive detection system was recommended for future work.
- Full Text:
- Date Issued: 1991
The pineal gland as a model to elucidate the primary mode of action of sympathoactive agents
- Authors: Welman, Alan David
- Date: 1991
- Subjects: Pineal gland , Cythochemistry
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4279 , http://hdl.handle.net/10962/d1002005 , Pineal gland
- Description: An attempt was made to use the pineal gland as a model for the study of the primary mode of action of sympathoactive agents. Two drugs were investigated, viz. alpha-methyldopa and ephedrine whose mode of action is not entirely clear. Organ cultures of pineal glands from rats treated chronically with alpha-methyldopa showed enhanced conversion of radioactive serotonin to melatonin (aMT) , as well as its precursor Nacetylserotonin (aHT). This treatment was also found to raise Nacetyltransferase (NAT) activity. These increases associated with alpha-methyldopa treatment were further enhanced by the beta-adrenergic agonist, isoproterenol, suggesting a supersensitivity-type effect occurring at the level of the beta-receptor. A subsequent binding study, however, showed a decrease in beta-receptor binding with exposure to alpha-methyldopa, providing mitigating evidence against the occurrence of a supersensitivity phenomenon. It is possible that a metabolite of alpha-methyldopa acts as an alpha 1 and beta-adrenergic agonist, resulting in greater melatonin (aMT) and N-acetylserotonin (aHT) synthesis than by a beta-adrenergic agonist, isoproterenol. Combined treatment of pineals with alpha-methyldopa and an alphareceptor blocker, phentolamine, resulted in melatonin (aMT) , Nacetylserotonin (aHT) , and N-acetyltransferase (NAT) activity levels which were lower than those obtained with alpha-methyldopa treatment alone, thus confirming the alpha-adrenergic activity of the metabolite of alpha-methyldopa. Additional pineal metabolites were isolated and measured simultaneously in the organ culture experiments. Organ cultures of rat pineal glands treated with ephedrine showed raised levels of melatonin (aMT) and N-acetylserotonin (aHT). Treatment with ephedrine also produced raised N-acetyltransferase activity. A further enhancement of these parameters was induced by norepinephrine, suggesting a supersensitivity-type effect occurring at the level of the beta-adrenergic receptor. Rats were treated with reserpine (a norepinephrine depleter) and the pineals exposed to ephedrine. Endogenous norepinephrine normally released by the action of ephedrine was thus absent, and under these conditions, levels of melatonin (aMT) and N-acetylserotonin (aHT) were reduced. N-acetyltransferase (NAT) activity was also reduced, but maintained levels pointing to substantial adrenergic activity of ephedrine as well as norepinephrine released by virtue of the drug's action. A subsequent binding study showed a decrease in beta-adrenergic receptor binding with exposure to ephedrine and a further decrease in ephedrine treated pineals from reserpine treated rats, thus ruling out the occurrence of a supersensitivity phenomenon. It is possible that both ephedrine and released norepinephrine have alpha- and beta-receptor activity. Additional pineal metabolites were isolated and measured in the organ culture experiments. A 16-hour time profile of the production of melatonin (aMT) and N-acetylserotonin (aHT) with norepinephrine and ephedrine treatment provided useful information regarding the course of action of the two agents. A pineal cell-culture system was developed and exposed to ephedrine and norepinephrine. N-acetyltransferase (NAT) activity levels measured after exposure to these agents were raised, confirming the adrenergic activity of both in the model. Finally, an HPLC system coupled to a UV detector was used in an attempt to measure melatonin (aMT) extracted from pineal organ culture media. The results showed that melatonin could be measured by this method, however, a more sensitive detection system was recommended for future work
- Full Text:
- Date Issued: 1991
A study of possible interactions between the pineal gland and the opioidergic system
- Authors: Khan, Razeeya B
- Date: 1990
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3729 , http://hdl.handle.net/10962/d1001468
- Description: Recent observations suggest a link between the pineal gland and the opioid system. Possible areas of interaction between the pineal gland and the opioidergic system in Wistar rats were investigated. The effect of opioids on the pineal gland in organ culture was monitored. Neither morphine, methadone nor the opioid antagonist naloxone was found to affect [¹⁴C]-serotonin metabolism by the pineal gland in vitro. Both the pineal gland and the opioid system are influenced by exposure to stressful stimuli. Morphine and melatonin had protective effects on stress-induced gastric lesions. The ability of melatonin to inhibit lesion formation was found not to be exerted via an opioidergic mechanism. Evidence has been obtained for a possible modulation of the stress response by the pineal gland . The opioid drugs are the most potent analgesic agents available. A possible interaction between the opioid system and the pineal gland in the modulation of the response to noxious stimuli was investigated. An intact pineal gland was found to be necessary for the manifestation of the nocturnally increased response of rats to noxious stimuli
- Full Text:
- Date Issued: 1990
A study of the effects of the pineal hormone, melatonin, on dopaminergic transmission in the central nervous system of rats
- Authors: Burton, Susan Frances
- Date: 1990
- Subjects: Dopaminergic mechanisms Melatonin Pineal gland -- Secretions Neural transmission Pineal gland Nervous system
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3726 , http://hdl.handle.net/10962/d1001463
- Description: Dopamine mechanisms in the central nervous system are important in the control of both normal and abnormal motor function. The recent observations in both animal and human studies, that melatonin, the principal hormone of the pineal gland, may have a role in the control of movement and the pathophysiology of movement disorders, have given rise to the concept that melatonin may have a modulatory influence on central dopaminergic neurotransmission. This study makes use of three animal behavioural models as well as a biochemical model of central dopaminergic function to further investigate the concept. Results from studies using the biochemical model, which investigated the effect of melatonin on dopamine and apomorphine stimulation of dopamine-sensitive adenylate cylase, suggest that melatonin is neither a competitive antagonist nor agonist at the D₁ receptor level, although the possibility of physiological stimulation or antagonism is not excluded. In behavioural studies, prior melatonin mg/kg administration (1 and 10 (8M) ip) inhibited apomorphine induced stereotypy and locomotor activity in normal rats, and apomorphine-induced rotational behaviour in 6-hydroxydopamine and quinolinic acid lesioned rats. The possibility that these results may have physiological significance is borne out by the observation that, under enviromental lighting conditions that are associated with raised endogeous melatonin levels, apomorphine- induced stereotypy and locomotor activity is attenuated. The general conclusion is that melatonin has an inhibitory influence on central nervous system dopaminergic function, suggesting therefore, that the pineal gland and melatonin may have a role in the pathophysiology and treatment of movement and behavioural disorders associated with dopaminergic dysfunction
- Full Text:
- Date Issued: 1990