10853851
Scutellarin protects against Aβ-induced learning and memory deficits in rats: involvement of nicotinic acetylcholine receptors and cholinesterase
Aim:To examine the protective effects of scutellarin (Scu) on rats with learning and memory deficit induced by β-amyloid peptide (Aβ).Methods:Fifty male Wistar rats were randomly divided into 5 groups: control, sham operation, Aβ, Aβ+Scu, and Aβ+piracetam groups. Aβ25–35 was injected into the lateral ventricle (10 μg each side). Scu (10 mg/2 mL) or piracetam (10 mg/2 mL was intragastrically administered per day for 20 consecutive days following Aβ treatment. Learning and memory was assessed with Morris water maze test. The protein and mRNA levels of nicotinic acetylcholine receptor (nAChR) α4, α7, and β2 subunits in the brain were examined using Western blotting and real-time PCR, respectively. The activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in the brain and plasma were measured using Ellman's colorimetric method.Results:In Aβ group, the escape latency period and first platform cross was significantly increased, and the total number of platform crossings was significantly decreased, as compared with the control and the sham operation groups. Both Scu and piracetam treatment significantly reduced the escape latency period and time to cross platform, and increased the number of platform crosses, but there were no significant differences between Aβ+Scu and Aβ+piracetam groups. In Aβ group, the protein levels of nAChR α4 and α7 subunits in the cerebral cortex were significantly decreased by 42%–47% and 58%–61%, respectively, as compared to the control and the sham operation groups. Scu treatment caused upregulation of α4 and α7 subunit proteins by around 24% and 30%, respectively, as compared to Aβ group, but there were no significant differences between Aβ+Scu and Aβ+piracetam groups. The protein level of nAChR β2 subunit had no significant difference among different groups. The mRNA levels of nAChR α4, α7, and β2 subunits were not significantly changed. In Aβ group, the activities of AChE and BuChE in the brain were significantly increased, but were significantly decreased in the plasma, as compared to the control and the sham operation groups. Scu or piracetam treatment restored the activities in brain and plasma nearly to the levels in the control group.Conclusion:The results suggest that Scu may rescue some of the deleterious effects of Aβ, possibly by stimulating nAChR protein translation and regulating cholinesterase activity.
45645511
Estrogenic and neuroprotective properties of scutellarin from Erigeron breviscapus: a drug against postmenopausal symptoms and Alzheimer's disease.
Besides the classical hormonal effect, estrogen possesses neuroprotective effects in the brain, which leads to the searching of novel treatments for neurodegenerative diseases such as Alzheimer's disease. Scutellarin is a major flavone derived from Herba Erigerontis, a Chinese medicine derived from Erigeron breviscapus, which has been shown here to possess both estrogenic and neuroprotective properties. Scutellarin showed the estrogenic effects by activating the estrogen responsive elements and phosphorylation of estrogen receptor alpha in cultured MCF-7 cells: the activation was in a dose-dependent manner. On the other hand, scutellarin inhibited the aggregation of beta-amyloid in vitro, and prevented the cell death mediated by beta-amyloid when applied to cultured neuronal PC12 cells. These results therefore suggested that Herba Erigerontis and its component scutellarin might have therapeutic effects against postmenopausal symptoms and Alzheimer's disease.
7845571
Commonly occurring plant flavonoids have estrogenic activity.
A remarkable diversity of naturally occurring and synthetic compounds have been shown to mimic the biological effects of 17 beta-estradiol by virtue of their ability to bind to and activate the nuclear estrogen receptor. This report extends the family of nonsteroidal estrogens to include several multiply hydroxylated chalcones, flavanones, and flavones. The hormone-like activity of these natural plant products is indicated by their ability to stimulate an estrogen receptor-dependent transcriptional response and to promote growth of estrogen-dependent MCF7 cells in culture. The transcriptional response can be inhibited by the steroidal estrogen antagonist ICI-164,384 and is specific for the estrogen receptor. Evidence is presented to show that selected hydroxylated flavonoids interact directly with the estrogen receptor, based on their ability to compete for the binding of 17 beta-[3H]estradiol to the receptor in cell-free extracts. These compounds are less active, on a molar basis, than 17 beta-estradiol or the synthetic dihydroxystilbene estrogens, but they have potencies comparable to those of other known phytoestrogens. Together, these findings broaden our understanding of the structure-activity relationships for nonsteroidal estrogens and present a series of new chemical prototypes for the future development of potentially useful agonists and antagonists for this nuclear receptor. The wide distribution of weakly estrogenic flavonoid pigments in food crops and medicinal plants raises additional questions about the possible health risks and benefits of these compounds, meriting closer examination of their presence in the human diet.
30089881
Flavonoids possess neuroprotective effects on cultured pheochromocytoma PC12 cells: a comparison of different flavonoids in activating estrogenic effect and in preventing beta-amyloid-induced cell death.
Despite the classical hormonal effect, estrogen possesses a neuroprotective effect in the brain, which has led many to search for novel treatments for neurodegenerative diseases. Flavonoids, a group of compounds mainly derived from vegetables, share a resemblance, chemically, to estrogen, and indeed, some have been used as estrogen substitutes. To search for potential therapeutic agents against neurodegenerative diseases, different subclasses of flavonoids were analyzed and compared with estrogen. First, the estrogenic activities of these flavonoids were determined by activating the estrogen-responsive elements in cultured MCF-7 breast cancer cells. Second, the neuroprotective effects of flavonoids were revealed by measuring its inhibition effects on the formation of reactive oxygen species, the aggregation of beta-amyloid, and the induction of cell death by beta-amyloid in cultured neuronal PC12 cells. Among these flavonoids, baicalein, scutellarin, hibifolin, and quercetin-3'-glucoside possessed the strongest effect in neuroprotection; however, the neuroprotective activity did not directly correlate with the estrogenic activity of the flavonoids. Identification of these flavonoids could be very useful in finding potential drugs, or food supplements, for treating Alzheimer's disease.
28426889
The Neuroprotective Effects of Phytoestrogens on Amyloid β Protein-induced Toxicity Are Mediated by Abrogating the Activation of Caspase Cascade in Rat Cortical Neurons*
Amyloid β protein (Aβ) elicits a toxic effect on neurons in vitro and in vivo. In present study we attempt to elucidate the mechanism by which Aβ confers its neurotoxicity. The neuroprotective effects of phytoestrogens on Aβ-mediated toxicity were also investigated. Cortical neurons treated with 5 μm Aβ-(25–35) for 40 h decreased the cell viability by 45.5 ± 4.6% concomitant with the appearance of apoptotic morphology. 50 μm kaempferol and apigenin decreased the Aβ-induced cell death by 81.5 ± 9.4% and 49.2 ± 9.9%, respectively. Aβ increased the activity of caspase 3 by 10.6-fold and to a lesser extent for caspase 2, 8, and 9. The Aβ-induced activation of caspase 3 and release of cytochromec showed a biphasic pattern. Apigenin abrogated Aβ-induced cytochrome c release, and the activation of caspase cascade. Kaempferol showed a similar effect but to a less extent. Kaempferol was also capable of eliminating Aβ-induced accumulation of reactive oxygen species. These two events accounted for the remarkable effect of kaempferol on neuroprotection. Quercetin and probucol did not affect the Aβ-mediated neurotoxicity. However, they potentiated the protective effect of apigenin. Therefore, these results demonstrate that Aβ elicited activation of caspase cascades and reactive oxygen species accumulation, thereby causing neuronal death. The blockade of caspase activation conferred the major neuroprotective effect of phytoestrogens. The antioxidative activity of phytoestrogens also modulated their neuroprotective effects on Aβ-mediated toxicity.
19178710
Estrogen for Alzheimer’s disease in women
Background: AD, the most prevalent cause of dementia, affects twice as many women as men. Therapeutic options are limited, but results of prior studies support the hypothesis that estrogen treatment may improve symptoms of women with this disorder. Methods: Forty-two women with mild-to-moderate dementia due to AD were enrolled into a randomized, double-blind, placebo-controlled, parallel-group trial of unopposed conjugated equine estrogens (1.25 mg/day) for 16 weeks. Results: Outcome data were available for 40 women at 4 weeks and 36 women at 16 weeks. At both 4 and 16 weeks, there were no significant differences or statistical trends between treatment groups on the primary outcome measure (the cognitive subscale of the Alzheimer’s Disease Assessment Scale), clinician-rated global impression of change, or caregiver-rated functional status. Exploratory analyses of mood and specific aspects of cognitive performance also failed to demonstrate substantial group differences. Conclusions: Although conclusions are limited by small sample size and the possibility of a type II error, results suggest that short-term estrogen therapy does not improve symptoms of most women with AD. These findings do not address possible long-term effects of estrogen in AD, possible interactions between estrogen and other treatment modalities, or putative effects of estrogen in preventing or delaying onset of this disorder.
46671204
Protective effects of scutellarin against cerebral ischemia in rats: evidence for inhibition of the apoptosis-inducing factor pathway.
Scutellarin (Scu) is the major active principle (flavonoid) extracted from Erigeron breviscapus (Vant.) Hand-Mazz, a Chinese herbal medicine. In this paper, we investigated the effects of Scu on brain injury through the inhibition of AIF-mediated apoptosis induced by transient focal brain ischemia in rats. Rats were treated with Scu for 7 d and then subjected to cerebral ischemia/reperfusion (I/R) injury induced by a middle cerebral artery occlusion (MCAO). After 2 h of ischemia and 22 h of reperfusion, the infarct volume and the neurological deficit were determined by TTC staining and Longa's score. IN SITU end-labeling of nuclear DNA fragments (TUNEL) was employed to determine the degree of DNA fragmentation. NAD content and PARP activity in brain homogenate were determined. The expression of AIF in the nucleus was analyzed by Western blot. The present study showed that Scu significantly reduced the infarct volume and ameliorated the neurological deficit. An increase in the number of TUNEL-positive cells and a decrease in the NAD level were also observed after 2 h of ischemia and 22 h of reperfusion. At the same time, Scu (50 and 75 mg kg (-1), i. g.) treatment reversed brain NAD depletion and reduced DNA fragmentation. Scu also inhibited PARP overactivation and AIF translocation from the mitochondria to the nucleus following cerebral I/R. These findings suggested that the neuroprotective effects of Scu on brain ischemic injury-induced apoptosis might be associated with inhibition of PARP-dependent mitochondrial dysfunction and subsequent translocation of AIF.
21184356
Neuronal nicotinic acetylcholine receptor–cholesterol crosstalk in Alzheimer's disease
Alzheimer's disease (AD) is one of the most devastating diseases of the central nervous system (CNS). It is characterized by two neuropathological findings: amyloid plaques and neurofibrillary tangles. AD is also accompanied by an extensive functional deficit in the cholinergic system, involving the neuronal‐type nicotinic acetylcholine receptor (AChR). Furthermore there is increasing evidence showing a misregulation of cholesterol metabolism in the development of the disease. Since cholesterol affects AChR protein at multiple levels, the cognitive impairment and other neurological correlates of AD might be partly associated with an abnormal crosstalk between the receptor protein and the sterol in this synaptopathy.
8544844
Intracellular Accumulation of Amyloid-Beta – A Predictor for Synaptic Dysfunction and Neuron Loss in Alzheimer's Disease
Despite of long-standing evidence that beta-amyloid (Aβ) peptides have detrimental effects on synaptic function, the relationship between Aβ, synaptic and neuron loss is largely unclear. During the last years there is growing evidence that early intraneuronal accumulation of Aβ peptides is one of the key events leading to synaptic and neuronal dysfunction. Many studies have been carried out using transgenic mouse models of Alzheimer's disease (AD) which have been proven to be valuable model systems in modern AD research. The present review discusses the impact of intraneuronal Aβ accumulation on synaptic impairment and neuron loss and provides an overview of currently available AD mouse models showing these pathological alterations.
1591081
Oxidative Stress Mediates the Pathogenic Effect of Different Alzheimer's Disease Risk Factors
Alzheimer's disease (AD) is a progressive neurodegenerative disorder affecting the elderly population. Mechanistically, the major cause of the disease bases on the altered processing of the amyloid-β (Aβ) precursor protein (APP), resulting in the accumulation and aggregation of neurotoxic forms of Aβ. Aβ derives from the sequential proteolytic cleavage of the β- and γ-secretases on APP. The causes of Aβ accumulation in the common sporadic form of AD are not completely known, but they are likely to include oxidative stress (OS). OS and Aβ are linked to each other since Aβ aggregation induces OS in vivo and in vitro, and oxidant agents increase the production of Aβ. Moreover, OS produces several effects that may contribute to synaptic function and cell death in AD. We and others have shown that the expression and activity of β-secretase (named BACE1; β-site APP cleaving enzyme) is increased by oxidant agents and by lipid peroxidation product 4-hydroxynonenal and that there is a significant correlation between BACE1 activity and oxidative markers in sporadic AD. OS results from several cellular insults such as aging, hyperglycemia, hypoxic insults that are all well known risk factors for AD development. Thus, our data strengthen the hypothesis that OS is a basic common pathway of Aβ accumulation, common to different AD risk factors.
42299206
In vivo conversion of racemized beta-amyloid ([D-Ser 26]A beta 1-40) to truncated and toxic fragments ([D-Ser 26]A beta 25-35/40) and fragment presence in the brains of Alzheimer's patients.
The lag between beta-amyloid (A beta) deposition and neurodegeneration in Alzheimer's disease (AD) suggests that age-dependent factors are involved in the pathogenesis. Racemization of Ser and Asp in A beta is a typical age-dependent modification in AD. We have shown recently that A beta1-40 racemized at Ser(26) ([D-Ser(26)]A beta 1-40) is soluble and non-toxic to neuronal cells, but is easily converted by brain proteases to truncated toxic fragments, [D-Ser(26)]A beta 25-35/40. Furthermore, [D-Ser(26)]A beta1-40 in vivo, produced a drastic and synergistic neuronal loss by enhancing the excitotoxicity when co-injected into rat hippocampus with ibotenic acid, an excitatory amino acid, suggesting an in vivo conversion of non-toxic [D-Ser(26)]A beta1-40 to toxic fragments including [D-Ser(26)]A beta 25-35/40. In this study, we further investigated the mechanism behind the in vivo neuronal loss by [D-Ser(26)]A beta1-40 and ibotenic acid in rats, and also searched for the presence of [D-Ser(26)]A beta 25-35/40 antigens in AD brains. Quantitative analyses of the damaged area indicate clearly that non-toxic [D-Ser(26)]A beta 1-40 caused as much neurodegeneration as toxic [D-Ser(26)]A beta 25-35/40. MK-801, an NMDA receptor antagonist, completely inhibited the neurodegeneration. The immunohistochemical analyses using anti-[D-Ser(26)]A beta 25-35/40-specific antibodies demonstrated the presence of [D-Ser(26)]A beta 25-35/40 antigens in senile plaques and in degenerating hippocampal CA1 neurons in AD brains, but not in age-matched control brains. These results strengthen our hypothesis that soluble [D-Ser(26)]A beta1-40, possibly produced during aging, is released from plaques and converted by proteolysis to toxic [D-Ser(26)]A beta 25-35/40, which damage hippocampal CA1 neurons by enhancing excitotoxicity in AD. This may account for the lag between A beta deposition and neurodegeneration in AD.
7688823
Acetylcholine in mind: a neurotransmitter correlate of consciousness?
The cholinergic system is one of the most important modulatory neurotransmitter systems in the brain and controls activities that depend on selective attention, which are an essential component of conscious awareness. Psychopharmacological and pathological evidence supports the concept of a 'cholinergic component' of conscious awareness. Drugs that antagonize muscarinic receptors induce hallucinations and reduce the level of consciousness, while the nicotinic receptor is implicated as being involved in the mechanism of action of general (inhalational) anaesthetics. In degenerative diseases of the brain, alterations in consciousness are associated with regional deficits in the cholinergic system. In Alzheimer's disease (AD), there is a loss of explicit (more than implicit) memory and hypoactivity of cholinergic projections to the hippocampus and cortex, while the visual hallucinations experienced by subjects with Dementia with Lewy bodies (DLB) are associated with reductions in neocortical ACh-related activity. In Parkinson's disease, the additional loss of pedunculopontine cholinergic neurones, which control REM (rapid eye movement) sleep or dreaming, is likely to contribute to REM abnormalities, which also occur in DLB. Widespread basal-forebrain and rostral brainstem cholinergic pathways, which include converging projections to the thalamus, appear to be located strategically for generating and integrating conscious awareness. Alleviation of a range of cognitive and non-cognitive symptoms by drugs that modulate the cholinergic system, which are being developed for the treatment of AD and related disorders, could be caused by changes in consciousness.
25627675
The cholinergic lesion of Alzheimer's disease: pivotal factor or side show?
A profound loss of cortical cholinergic innervation is a nearly invariant feature of advanced Alzheimer's disease (AD). The temporal course of this lesion and its relationship to other aspects of the disease have not yet been fully clarified. Despite assertions to the contrary, a review of the evidence suggests that a perturbation of cholinergic innervation is likely to be present even in the very early stages of AD. This cholinergic lesion is unlikely to be a major determinant of the clinical symptoms or of the neuropathological lesions. Nonetheless, it almost certainly contributes to the severity of the cognitive and behavioral deficits, especially in the areas of memory and attention. The cholinergic lesion may also influence the progression of the neuropathological process through complex interactions with amyloidogenesis, tau phosphorylation and neuroplasticity.
27207955
Neuronal nicotinic receptors in the human brain
Neuronal nicotinic acetylcholine receptors (nAChRs) are a family of ligand gated ion channels which are widely distributed in the human brain. Multiple subtypes of these receptors exist, each with individual pharmacological and functional profiles. They mediate the effects of nicotine, a widely used drug of abuse, are involved in a number of physiological and behavioural processes and are additionally implicated in a number of pathological conditions such as Alzheimer's disease, Parkinson's disease and schizophrenia. The nAChRs have a pentameric structure composed of five membrane spanning subunits, of which nine different types have thus far been identified and cloned. The multiple subunits identified provide the basis for the heterogeneity of structure and function observed in the nAChR subtypes and are responsible for the individual characteristics of each. A substantial amount of information on human nAChR structure and function has come from studies on neuroblastoma cell lines which naturally express nAChRs and from recombinant nAChRs expressed in Xenopus oocytes. In vitro brain nAChR distribution can be mapped with a number of appropriate agonist and antagonist radioligands and subunit distribution may be mapped by in situ hybridization using subunit specific mRNA probes. Receptor distribution in the living human brain can be studied with noninvasive imaging techniques such as PET and SPECT, with a significant reduction in nAChRs in the brains of Alzheimer's patients having been identified with [11C] nicotine in PET studies. Despite the significant body of knowledge now accumulated about nAChRs, much remains to be elucidated. This review will attempt to describe the current knowledge on the nAChR subtypes in the human brain, their functional roles and neuropathological involvement.
29238038
New anti-Alzheimer drugs from biodiversity: the role of the natural acetylcholinesterase inhibitors.
Alzheimer's disease (AD) is a progressive neurodegenerative pathology with severe economic and social impact. There is currently no cure, although cholinesterase inhibitors provide effective temporary relief of symptoms in some patients. Nowadays, drug research and development are based on the cholinergic hypothesis that supports the cognition improvement by regulation of the synthesis and release of acetylcholine in the brain. There are only four commercial medicines approved for treatment of AD, and natural products have played an important alternative role in the research for new acetylcholinesterase inhibitors, as exemplified through the discovery of galantamine. This profile conducts us to give in this paper an overview relating the several classes of natural products with anti-cholinesterasic activity as potential templates to the design of new selective and powerful anti-Alzheimer drugs.
13795016
Tetrahydrofurobenzofuran cymserine, a potent butyrylcholinesterase inhibitor and experimental Alzheimer drug candidate, enzyme kinetic analysis
Synaptic loss, particularly related to the forebrain cholinergic system, is considered to be an early event that leads to Alzheimer’s disease (AD) and has led to the development of acetylcholinesterase inhibitors (AChE-Is) as the mainstay of treatment for several degenerative disorders that culminate in dementia. The primary dose-limiting toxicities of all clinically available AChE-Is are, similar to useful actions on cognition, cholinergically mediated and they ultimately limit the value of this drug class in achieving anything but symptomatic improvements. In addition, AChE levels in brain areas associated with AD decline with disease progression, which likely ultimately limits the therapeutic utility of this drug class. New research indicates that selective inhibition of butyrylcholinesterase (BuChE), a closely related enzyme that is markedly elevated in AD brain, increases acetylcholine (ACh) and augments cognition in rodents free of the characteristic undesirable actions of AChE-Is. BuChE inhibition hence represents an innovative treatment approach for AD, and agents are currently being synthesized to optimally achieve this. The novel compound, tetrahydrofurobenzofuran cymserine (THFBFC), is derived from our effort to produce a potent and BuChE-selective inhibitor as a candidate to test the hypothesis that BuChE-Is would be efficacious and better tolerated than AChE-Is in AD. Herein, we applied innovative enzyme kinetic analyses to characterize the quantitative interaction of THFBFC with human BuChE. These provided values for the agent’s IC50, together with specific new kinetic constants, such as KT50, KT1/2, RI, oKRT, oPmax, KPT and PT1/2, to aid define target concentrations for clinical translation. Additional classical kinetic parameters, including Ki, Km or Ks, kcat or Vmax and Vmi were also determined. THFBFC proved to be a potent competitive inhibitor of human BuChE and, like its isomer dihydrobenzodioxepine cymserine, is a potentially interesting AD drug candidate.
40160706
Amnesia induced by beta-amyloid fragments is counteracted by cannabinoid CB1 receptor blockade.
Administration of drugs activating cannabinoid CB(1) receptors in the brain induces memory deficit in rodents, and blockade of these receptors may restore memory capacity in these animals. Central administration of beta-amyloid or beta-amyloid fragments may also lead to memory disturbances. This study was undertaken to study the involvement of cannabinoid CB(1) receptors in amnesia induced by beta-amyloid fragments in mice tested in a step-through passive avoidance paradigm. Pre-training intracerebroventricular (i.c.v.) injection of beta-amyloid fragments, beta-amyloid peptide-(25-35) (4, 8 or 16 nmol/mouse) or beta-amyloid peptide-(1-42) (200, 400, 800 pmol/mouse) 7 days prior to the learning trial reduced in a dose-dependent manner the retention of passive avoidance response. This effect was observed in two retention tests, 1 and 7 days after the learning trial. The two beta-amyloid fragments showed similar potency in reducing retention of passive avoidance behavior. This effect was counteracted by a single intraperitoneal (i.p.) injection of the cannabinoid CB(1) receptor antagonist, N-(piperidin-l-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride (SR141716A, 1 mg/kg), made 30 min prior to the second retention test. The injection of SR141716A per se did not affect memory capacity of mice. The i.c.v. administration of beta-amyloid peptide-(25-35) (8 nmol/mouse) or of beta-amyloid peptide-(1-42) (400 pmol/mouse) made 30 min prior to the learning trial failed to affect the retention capacity of mice as measured 1 and 7 days later. Also, the i.p. injection of SR 141716A (1 mg/kg) made 30 min prior to the learning trial did not influence the behavioral response of mice injected with beta-amyloid peptide-(25-35) (8 nmol/mouse) or of beta-amyloid peptide-(1-42) (400 pmol/mouse) 7 days prior to the learning trial. These results show that beta-amyloid fragments induce a dose-dependent memory deficit. Their effect on memory retention depends upon the time of administration and seems to involve cannabinoid CB(1) receptors in the brain.
28426889
The Neuroprotective Effects of Phytoestrogens on Amyloid β Protein-induced Toxicity Are Mediated by Abrogating the Activation of Caspase Cascade in Rat Cortical Neurons*
Amyloid β protein (Aβ) elicits a toxic effect on neurons in vitro and in vivo. In present study we attempt to elucidate the mechanism by which Aβ confers its neurotoxicity. The neuroprotective effects of phytoestrogens on Aβ-mediated toxicity were also investigated. Cortical neurons treated with 5 μm Aβ-(25–35) for 40 h decreased the cell viability by 45.5 ± 4.6% concomitant with the appearance of apoptotic morphology. 50 μm kaempferol and apigenin decreased the Aβ-induced cell death by 81.5 ± 9.4% and 49.2 ± 9.9%, respectively. Aβ increased the activity of caspase 3 by 10.6-fold and to a lesser extent for caspase 2, 8, and 9. The Aβ-induced activation of caspase 3 and release of cytochromec showed a biphasic pattern. Apigenin abrogated Aβ-induced cytochrome c release, and the activation of caspase cascade. Kaempferol showed a similar effect but to a less extent. Kaempferol was also capable of eliminating Aβ-induced accumulation of reactive oxygen species. These two events accounted for the remarkable effect of kaempferol on neuroprotection. Quercetin and probucol did not affect the Aβ-mediated neurotoxicity. However, they potentiated the protective effect of apigenin. Therefore, these results demonstrate that Aβ elicited activation of caspase cascades and reactive oxygen species accumulation, thereby causing neuronal death. The blockade of caspase activation conferred the major neuroprotective effect of phytoestrogens. The antioxidative activity of phytoestrogens also modulated their neuroprotective effects on Aβ-mediated toxicity.
17544260
Beta-amyloid (1-42)-induced learning and memory deficits in mice: involvement of oxidative burdens in the hippocampus and cerebral cortex.
We have demonstrated that oxidative stress is involved, at least in part, in beta-amyloid protein (Abeta)-induced neurotoxicity in vivo [Eur. J. Neurosci. 1999;11:83-90; Neuroscience 2003;119:399-419]. However, mechanistic links between oxidative stress and memory loss in response to Abeta remain elusive. In the present study, we examined whether oxidative stress contributes to the memory deficits induced by intracerebroventricular injection of Abeta (1-42) in mice. Abeta (1-42)-induced memory impairments were observed, as measured by the water maze and passive avoidance tests, although these impairments were not found in Abeta (40-1)-treated mice. Treatment with antioxidant alpha-tocopherol significantly prevented memory impairment induced by Abeta (1-42). Increased activities of the cytosolic Cu,Zn-superoxide dismutase (Cu,Zn-SOD) and mitochondrial Mn-superoxide dismutase (Mn-SOD) were observed in the hippocampus and cerebral cortex of Abeta (1-42)-treated animals, as compared with Abeta (40-1)-treated mice. The induction of Cu,Zn-SOD was more pronounced than that of Mn-SOD after Abeta (1-42) insult. However, the concomitant induction of glutathione peroxidase (GPX) in response to significant increases in SOD activity was not seen in animals treated with Abeta (1-42). Furthermore, glutathione reductase (GRX) activity was only increased at 2h after Abeta (1-42) injection. Production of malondialdehyde (lipid peroxidation) and protein carbonyl (protein oxidation) remained elevated at 10 days post-Abeta (1-42), but the antioxidant alpha-tocopherol significantly prevented these oxidative stresses. Therefore, our results suggest that the oxidative stress contributes to the Abeta (1-42)-induced learning and memory deficits in mice.
30946789
Suppressed expression of nicotinic acetylcholine receptors by nanomolar β-amyloid peptides in PC12 cells
Summary. A line of evidence has shown that a link between the common pathological features of β-amyloid peptide (Aβ) deposition and cholinergic degeneration observed in Alzheimer's disease (AD) may exist, however, no experimental evidence has shown that exposure to Aβ can decrease expression of nicotinic acetylcholine receptors (nAChRs), which have been shown to play roles in brain cognitive functions. Here, we report that treatment with Aβ1–40 and Aβ25–35 at nanomolar concentrations significantly decreased the [3H]epibatidine and [125I]α-bungarotoxin binding sites, the protein and mRNA levels of nAChR α3, α7 and β2 subunits in PC12 cells. Aβ1–40 and Aβ25–35 at the concentrations used in the treatment study neither bound to nAChRs nor induced apoptosis, but significantly inhibited the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5,diphenyl tetrazolium bromide) reduction. These data suggest that the decreased biosynthesis of nAChRs induced by Aβ may be attributable partially to perturbances of some intracellular signal transduction pathways. The results presented in this study lead to a hypothesis that Aβ can degenerate nAChRs early in the course of AD before the formation of abundant Aβ fibrils.
12695564
Nicotinic Acetylcholine Receptor Signalling: Roles in Alzheimer's Disease and Amyloid Neuroprotection
Alzheimer's disease (AD), the major contributor to dementia in the elderly, involves accumulation in the brain of extracellular plaques containing the β-amyloid protein (Aβ) and intracellular neurofibrillary tangles of hyperphosphorylated tau protein. AD is also characterized by a loss of neurons, particularly those expressing nicotinic acetylcholine receptors (nAChRs), thereby leading to a reduction in nAChR numbers. The Aβ1–42 protein, which is toxic to neurons, is critical to the onset and progression of AD. The discovery of new drug therapies for AD is likely to be accelerated by an improved understanding of the mechanisms whereby Aβ causes neuronal death. We examine the evidence for a role in Aβ1–42 toxicity of nAChRs; paradoxically, nAChRs can also protect neurons when activated by nicotinic ligands. Aβ peptides and nicotine differentially activate several intracellular signaling pathways, including the phosphatidylinositol 3-kinase/v-akt murine thymoma viral oncogene homolog pathway, the extracellular signal-regulated kinase/mitogen-activated protein kinase, and JAK-2/STAT-3 pathways. These pathways control cell death or survival and the secretion of Aβ peptides. We propose that understanding the differential activation of these pathways by nicotine and/or Aβ1–42 may offer the prospect of new routes to therapy for AD.
37910714
Nicotinic acetylcholine receptors and nicotinic cholinergic mechanisms of the central nervous system.
Subtypes of neuronal nicotinic acetylcholine receptors (nAChRs) are constructed from numerous subunit combinations that compose channel-receptor complexes with varied functional and pharmacological characteristics. Structural and functional diversity and the broad presynaptic, postsynaptic, and nonsynaptic locations of nAChRs underlie their mainly modulatory roles throughout the mammalian brain. Presynaptic and preterminal nicotinic receptors enhance neurotransmitter release, postsynaptic nAChRs contribute a small minority of fast excitatory transmission, and nonsynaptic nAChRs modulate many neurotransmitter systems by influencing neuronal excitability. Nicotinic receptors have roles in development and synaptic plasticity, and nicotinic mechanisms participate in learning, memory, and attention. Decline, disruption, or alterations of nicotinic cholinergic mechanisms contribute to dysfunctions such as epilepsy, schizophrenia, Parkinson's disease, autism, dementia with Lewy bodies, Alzheimer's disease, and addiction.
23066313
Nicotinic acetylcholine receptors of muscles and nerves: comparison of their structures, functional roles, and vulnerability to pathology.
There are fetal and adult subtypes of muscle nicotinic receptors (AChRs), whose structures and functional roles are reasonably well known. Mutations of their subunits cause congenital myasthenic syndromes. An autoimmune response to them causes myasthenia gravis (MG). The main immunogenic region (MIR) on muscle AChRs accounts for many aspects of the pathological mechanisms by which the autoimmune response impairs neuromuscular transmission. There are many other AChR subtypes, each defined by a different combination of subunits, some of which are transiently expressed in muscle during development, others of which are expressed in keratinocytes, vascular and bronchial epithelia, and other nonneuronal cells, as well as in a wide variety of neurons. Their varied structures and functional roles are much less well known. Mutations in subunits of some of these AChRs have thus far been associated with rare forms of epilepsy and dysautonomia, but other genetic diseases associated with them probably remain to be discovered. Autoimmune responses to some of these subunits are associated with rare dysautonomias and a skin disease. The pathological mechanisms by which these autoimmune responses impair function are much less well known than in the case of MG. AChRs may provide useful drug targets in several neurological diseases. By far, the biggest direct medical impact of AChRs is addiction to tobacco, which is mediated by nicotine acting on a variety of neuronal AChRs.
24996503
Decreased Protein Levels of Nicotinic Receptor Subunits in the Hippocampus and Temporal Cortex of Patients with Alzheimer’s Disease
Abstract: Deficits of cortical nicotinic acetylcholine receptors (nAChRs) have been observed in Alzheimer’s disease (AD) by receptor binding assays. Little is known about the receptor subunit specificity influenced by AD, and it might be of importance for therapeutic strategies. In the present study, the protein levels of nAChR α3, α4, α7, and β2 subunits were investigated using western blot analysis on postmortem brains of patients with AD and age‐matched controls. The results showed that in human postmortem brain samples, bands with molecular masses of 52, 42, and 50 kDa were detected by anti‐α4, anti‐α7, and anti‐β2 antibodies, respectively. When anti‐α3 antibody was used, one major band of 49 kDa and two minor bands of 70 and 38 kDa were detected. In AD patients, as compared with age‐matched controls, the α4 subunit was reduced significantly by ∼35 and 47% in the hippocampus and temporal cortex, respectively. A significant reduction of 25% in the α3 subunit was also observed in the hippocampus and a 29% reduction in the temporal cortex. For the α7 subunit, the protein level was reduced significantly by 36% in the hippocampus of AD patients, but no significant change was detected in the temporal cortex. In neither the hippocampus nor the temporal cortex was a significant difference observed in the β2 subunit between AD patients and controls. These results reveal brain region‐specific changes in the protein levels of the nAChR, α3, α4, and α7 subunits in AD.
28937549
Selective changes in the levels of nicotinic acetylcholine receptor protein and of corresponding mRNA species in the brains of patients with Parkinson’s disease
Reductions in the number of neuronal nicotinic acetylcholine receptors (nAChRs) have been shown to occur in connection with Parkinson's disease (PD), but it is still unclear which subtype of this receptor is affected. In the present study we examined various nAChR subtypes employing ligand binding, as well as levels of subunit protein and mRNA in the brains of PD patients and age-matched controls. Binding of [3H]epibatidine and levels of alpha3 mRNA in the caudate nucleus and temporal cortex, but not in the hippocampus were significantly decreased in the PD brain. The level of the alpha3 protein subunit was significantly reduced in all these brain regions but there was no change in the level of alpha4. The level of the beta2 protein subunit in the temporal cortex and hippocampus and the beta2 mRNA in the temporal cortex was lowered. Both the levels of the alpha7 subunit protein and [125I]alpha-bungarotoxin binding were significantly increased in the temporal cortex of PD patients whereas the alpha7 mRNA level was unchanged. These findings reveal selective losses of the alpha3- and beta2-containing nAChRs and an increase in the alpha7 nAChRs that might be related to the pathogenesis of PD.
17744682
Nicotinic receptor abnormalities in Alzheimer’s disease
Loss of cortical nicotinic acetylcholine receptors with high affinity for agonists (20-50%) in patients with Alzheimer's disease is a common finding. Recent immunochemical analyses indicate that this deficit is predominantly associated with the loss of alpha4 subunits (30-50%), although modest reductions of alpha3 may occur in some individuals (25-29%). No reduction of beta2 subunit protein expression or levels of alpha3 and alpha4 messenger RNA has been reported. Decline in cortical [(125)I]alpha-bungarotoxin binding and alpha7 protein expression does not appear to be as extensive or widespread as the loss of alpha4 (0-40%), with no reduction in messenger RNA expression. In the thalamus, there was a trend for reduced [(3)H]nicotine binding in the majority of nuclei (0-20%) in Alzheimer's disease; however, there was a significant decline in [(125)I]alpha-bungarotoxin binding in the reticular nucleus. In the striatum [(3)H]nicotine binding was reduced in Alzheimer's disease, and although neuroleptic medication accentuated this change, it occurred in those free of neuroleptics. Changes in nicotinic acetylcholine receptors in Alzheimer's disease are distinct from those in normal aging and are likely to contribute to clinical features and possibly neuropathology.
86211914
The Rat Brain in Stereotaxic Coordinates
Preface. Acknowledgements. Introduction. References. List of Structures. Index of Abbreviations. Diagrams.
1473315
Decreased nicotinic receptors and cognitive deficit in rats intracerebroventricularly injected with beta‐amyloid peptide(1‐42) and fed a high‐cholesterol diet
To investigate whether the changes in nicotinic receptors (nAChRs) and in learning and memory associated with Alzheimer's disease (AD) are influenced by both β‐amyloid peptide (Aβ) and cholesterol in vivo, we examined the effects of intracerebroventricular injection of Aβ1–42 and/or a high‐cholesterol diet on brain levels ofnAChRs and learning and memory in rats. The levels of nAChR subunit proteins and the corresponding mRNA were measured by Western blotting and RT‐PCR, respectively; and learning and memory were evaluated with the Morris Water Maze examination. Injection of Aβ1–42 resulted in deposition of this peptide, activation of astrocytes, decreased levels of the α7 and α4 protein subunits of the nAChR, and elevated expression of α7 mRNA, as well as impaired learning and spatial memory. A high‐cholesterol diet activated astrocytes and, more importantly, potentiated the toxic effects of Aβ on nAChR subunit levels and on learning and memory. These findings may be highly relevant to the mechanisms underlying the cognitive deficits associated with AD. © 2007 Wiley‐Liss, Inc.
2230493
Amyloid precursor protein gene mutated at Swedish 670/671 sites in vitro induces changed expression of nicotinic acetylcholine receptors and neurotoxicity
In order to investigate the influence of amyloid precursor protein (APP) over-expression on the levels of nicotinic acetylcholine receptors (nAChRs), the pCDNA 3.0 carrying the Swedish 670/671 APP double mutation (APP(SWE)) gene was transfected into human neuroblastoma (SH-SY5Y) cells and primary culture of rat hippocampal neurons. The mRNA level of APP, and nAChR α3, α4 and α7 subunits were detected by real-time PCR, and their corresponding proteins as well as α-secreted APP (αAPPs) by Western blotting. [3H]Epibatidine binding sites were measured by the receptor binding assay. The results showed that significantly concomitant increases in mRNA and protein levels of SH-SY5Y cells and primary cultured neurons transfected with APP(SWE) were observed. Interestingly, a decreased αAPPs level was detected in both cells treated with APP(SWE) transfection. In addition, decreases in mRNA and protein levels of α3 nAChR subunit in SH-SY5Y cells or α4 subunit in primary cultured neurons with APP(SWE) transfection were observed. For α7 nAChR, the increased protein and mRNA levels were found in SH-SY5Y cells and primary cultured neurons with APP(SWE) transfection. The number of cholinergic receptor binding site of [3H]epibatidine was decreased in the SH-SY5Y cells transfected with APP(SWE). Elevations in the activities of AChE and BuChE and in the level of lipid peroxidation were detected in both types of cultured cells transfected with APP(SWE). These results indicated that the over-expression of APP(SWE) gene can influence the expression of nAChRs and resulted in neurotoxicity, in which this process might play an important role in the pathogenesis of Alzheimer's disease.
11072006
A new and rapid colorimetric determination of acetylcholinesterase activity.
Abstract A photometric method for determining acetylcholinesterase activity of tissue extracts, homogenates, cell suspensions, etc., has been described. The enzyme activity is measured by following the increase of yellow color produced from thiocholine when it reacts with dithiobisnitrobenzoate ion. It is based on coupling of these reactions: The latter reaction is rapid and the assay is sensitive (i.e. a 10 μ1 sample of blood is adequate). The use of a recorder has been most helpful, but is not essential. The method has been used to study the enzyme in human erythrocytes and homogenates of rat brain, kidney, lungs, liver and muscle tissue. Kinetic constants determined by this system for erythrocyte eholinesterase are presented. The data obtained with acetylthiocholine as substrate are similar to those with acetylcholine.
18812400
Alzheimer's disease and acetylcholine receptors.
Cholinergic abnormalities, alongside senile plaques, neurofibrillary tangles, and extensive neuronal loss, are the major characteristics in Alzheimer's disease (AD). Both nicotinic and muscarinic acetylcholine receptors are decreased in AD, and it has been shown that the reduction in the number of acetylcholine receptors precedes other pathologic changes. Anti-cholinergic drugs induce amnesia, which can be reversed by withdrawal of the medication. Inhibition of the down-regulation of acetylcholine is, therefore, a strategy for the treatment of AD because it could augment acetylcholine levels within synaptic clefts. In this context, acetylcholinesterase inhibitors, which improve cognitive functions, are currently approved for the treatment of AD. Stimulation of acetylcholine receptors, nicotinic or muscarinic, is another strategy; some drugs are currently under investigation, and reported to be effective. In addition, nicotinic stimulation exerts a neuroprotective effect, and reduces the amyloid burden. Cholinergic therapy may counter the symptoms and progress of AD.
36066562
Acetylcholinesterase and butyrylcholinesterase--important enzymes of human body.
The serine hydrolases and proteases are a ubiquitous group of enzymes that is fundamental to many critical life-functions. Human tissues have two distinct cholinesterase activities: acetylcholinesterase and butyrylcholinesterase. Acetylcholinesterase functions in the transmission of nerve impulses, whereas the physiological function of butyrylcholinesterase remains unknown. Acetylcholinesterase is one of the crucial enzymes in the central and peripheral nerve system. Organophosphates and carbamates are potent inhibitors of serine hydrolases and well suited probes for investigating the chemical reaction mechanism of the inhibition. Understanding the enzyme's chemistry is essential in preventing and/or treating organophosphate and carbamate poisoning as well as designing new medicaments for cholinergic-related diseases like as Alzheimer's disease.
40596092
Cholinesterases within neurofibrillary tangles related to age and Alzheimer's disease
Enzyme histochemistry showed that a substantial number of cortical neurofibrillary tangles contain acetylcholinesterase and butyrylcholinesterase activities in patients with Alzheimer's disease and in nondemented aged individuals. In some areas of the brain, tangles containing each of these two enzymes were differentially distributed. In other areas, there was a very close overlap between the two types of cholinesterase.
33271083
Senile plaques: staining for acetylcholinesterase and A4 protein: a comparative study in the hippocampus and entorhinal cortex
SummaryIn 20 unselected autopsy cases tissue blocks from the hippocampus with adjacent entorhinal cortex and neocortex were stained for acetylcholinesterase (AChE). From five brains shown to have large numbers of senile plaques tissue, adjacent to that taken for AChE tissue blocks, was embedded in paraffin and sections were immunostained for the A4 protein. The morphological aspects were compared. Equivalent types of plaques and plaque-like structures were observed in the A4- and ACHE-stained sections. On selected tissue blocks from patients with many senile plaques two immediately adjacent cryostat sections were stained, one for AChE and one for A4 protein. The same individual plaques could be identified on the two sections. These findings suggest that high AChE activity is intimately associated with the process of A4 protein formation and accumulation in plaques and that this association already occurs at a very carly stage of plaque formation.
41766593
Therapies for Alzheimer's disease
Alzheimer’s disease (AD) is a devastating neurological disorder that affects more than 37 million people worldwide1. The economic burden of AD is massive; in the United States alone, the estimated direct and indirect annual cost of patient care is at least US$100 billion. However, the number of therapeutic options for AD remains severely limited. Currently marketed drugs for AD do not prevent or reverse this disease and are approved only for the management of symptoms. Driven by the clear unmet medical need and a better understanding of the biology and pathophysiology of AD, the number of drugs in development for this indication has increased dramatically in recent years.
12831211
Plants used in Chinese and Indian traditional medicine for improvement of memory and cognitive function
In traditional practices of Ayurvedic and Chinese medicine, numerous plants have been used to treat cognitive disorders, including neurodegenerative diseases such as Alzheimer's disease (AD). An ethnopharmacological approach has provided leads to identifying potential new drugs from plant sources, including those for cognitive disorders. Many drugs currently available in Western medicine were originally isolated from plants, or are derived from templates of compounds isolated from plants. Some anticholinesterase (anti-ChE) alkaloids isolated from plants have been investigated for their potential in the treatment of AD, and are now in clinical use. Galantamine, isolated from several plants including Lycoris radiata Herb., which was used in traditional Chinese medicine (TCM), is licensed in the United Kingdom for the treatment of mild to moderate AD. Various other plant species have shown pharmacological activities relevant to the treatment of cognitive disorders, indicating potential for therapeutic use in disorders such as AD. This article reviews some of the plants and their active constituents that have been used in traditional Ayurvedic medicine and TCM for their reputed cognitive-enhancing or antiageing effects. Plants and their constituents with pharmacological activities that may be relevant for the treatment of cognitive disorders, including enhancement of cholinergic function in the central nervous system (CNS), anti-inflammatory and antioxidant activities, are discussed.
42076209
Breviscapine improves functions of spatial learning and memory of focal cerebral ischemia rats.
OBJECTIVE
To investigate the effects of breviscapine on the functions of spatial learning and memory of focal cerebral ischemia rats.


METHODS
Rots withl the left middle cerebral artery occluded were made by an intraluminal filament. Then breviscapine (20 mg/kg,40 mg/kg) in experimental group and 10% glucose in control group were administered intraperitoneally once a day for 2 weeks, and the Morris water maze tasks were carried out for 5 days.


RESULTS
Compared with sham-operation group,the animals of ischemia-control group exhibited seriously impaired spatial learning and memory in both place navigation test and spatial probe test. In the place navigation test, the mean value of escape latency in breviscapine group was significantly shorter than that in ischemia control group (P < 0.01 for lower-dose and P < 0.05 for higher-dose breviscapine group, respectively). In the spatial probe test,compared with sham-operation (P < 0.01) and breviscapine group (P < 0.01), the rats of ischemia-control group spent more time in the no-former platform quadrant, and showed reduced frequency of crossing former platform site significantly. The numbers of neurons with Nissl staining and choline acetyltransferase (ChAT) immunopositive neurons in ipssilateral cortex in the breviscapine group were significantly more than those in the ischemia-control group (P < 0.01). In hippocampus, the numbers of neurons with Nissl staining and ChAT immunopositive neurons in the stratum pyramidale of CA area were similar among the groups.


CONCLUSION
These results indicate that breviscapine can improve the functions of spatial learning and memory of focal cerebral ischemia rats and the protection against the loss of ChAT immunopositive neuron in new cortex may be involved in its mechanisms.
1344728
Protective effects of breviscapine on ischemic vascular dementia in rats.
Breviscapine, a traditional Chinese medicine, is extensively used in clinic to treat cardiovascular diseases and cerebrovascular injury. In this study, we demonstrated the effects of breviscapine on vascular dementia (VD) rats, which were mimicked by permanent occlusion of bilateral common carotid arteries. Breviscapine (2 mg/kg for 14 d) improved the performance of learning and memory of VD rats in Morris water maze, decreased the level of lipid peroxidation and free radicals, and attenuated the pathological alterations, such as nuclear shrink, cellular edema and irregular arrangement of pyramidal layer in the hippocampal CA(1) area. In vitro experiment, breviscapine (50 microg/l) protected cortical neuron from injury and decreased intracellular calcium overloading induced by H2O2 (10 mM). The results suggest that breviscapine has therapeutic effect on cerebral ischemia and vascular dementia.
36331548
Protective effects of scutellarin on superoxide-induced oxidative stress in rat cortical synaptosomes.
AIM
To evaluate the effects of scutellarin on superoxide-induced oxidative stress in rat cortical synaptosomes.


METHODS
Oxidative damage model was established by incubation with xanthine (0.3 mmol/L) and xanthine oxidase (0.02 U) at 37 degree for 30 min. The extent of membrane oxidation was assessed by malondialdehyde (MDA). Membrane fluidity was measured by fluorescence anisotropy (polarization) of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene (DPH). Intracellular Ca2+ concentration([Ca2+]i) was measured by fluorescent spectrophotometry. Fura 2-AM was used as an indicator for [Ca2+]i. Na+/K+-ATPase activity assay was based on the amount of inorganic phosphate (Pi) released during an enzymatic hydrolysis of ATP.


RESULTS
Synaptosomes exposed to superoxide significantly elevated the levels of malondialdehyde (MDA) and [Ca2+]i compared with those in normal group. These changes were accompanied by the decrease in membrane fluidity and Na+/K+-ATPase activity. Pretreatment with scutellarin (25-100 micromol/L) significantly ameliorated the oxidative damage of synaptosomes by reducing MDA levels and [Ca2+]i, up-regulating membrane fluidity and restoring Na+/K+-ATPase activity.


CONCLUSION
Scutellarin exerts a potent protective effect against oxidative damage in synaptosomes induced by superoxide.
2854976
Effects of scutellarin on PKCγ in PC12 cell injury induced by oxygen and glucose deprivation
AbstractAim:To evaluate the neuroprotective effect and mechanisms of scutellarin (Scu) against PC12 cell injury after oxygen and glucose deprivation followed by reperfusion (OGD-Rep).Methods:Undifferentiated rat pheochromocytoma PC12 cells, exposed to oxygen and glucose deprivation followed by reperfusion (OGD-Rep), used as an in vitro model of ischemia/reperfusion. Cell survival was evaluated by diphenyltetrazolium bromide (MTT) assay and the amount of LDH release was determined using assay kits. [Ca2+]i was monitored using a fluorescent Ca2+-sensitive dye Fura-2 acetoxymethyl ester. Cell apoptosis was detected by a DNA ladder and by flow cytometric detection. The expression of protein kinase C (PKC)γ was determined using both RT-PCR and Western blotting. The translocation of PKCγ was assayed by subcellular fractionation and Western blotting.Results:OGD-Rep injury significantly elevated the level of LDH release, [Ca2+]i, mRNA expression and the translocation of PKCγ compared in the PC12 cells with those of the normal group. Scu (10-100 μmol/L) exerted a protective effect against OGD-Rep injury by reducing LDH release, [Ca2+]i, the percent of apoptosis, and the translocation of PKCγ.Conclusion:Scu inhibits the increase of [Ca2+]i and the activation of PKCγ, exerting protective effects against PC12 cell injury induced by OGD-Rep.
33866031
Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin.
The present study investigated the protective actions of the antioxidant scutellarin against the cytotoxicity produced by exposure to H2O2 in PC12 cells. This was done by assaying for MTT (3,(4,5-dimethylthiazole-2-yl)2,5-diphenyl-tetrazolium bromide) reduction and lactate dehydrogenase (LDH) release. Reactive oxygen species (ROS) and Ca2+ in cells were evaluated by fluorescent microplate reader using DCFH and Fura 2-AM, respectively, as probes. Lipid peroxidation was quantified using thiobarbituric acid-reactive substances (TBARS). Mitochondrial membrane potential (MMP) was assessed by the retention of rhodamine123 (Rh123), a specific fluorescent cationic dye that is readily sequestered by active mitochondria, depending on their transmembrane potential. The DNA content and percentage of apoptosis were monitored with flow cytometry. Vitamin E, a potent antioxidant, was employed as a comparative agent. Preincubation of PC12 cells with scutellarin prevented cytotoxicity induced by H2O2. Intracellular accumulation of ROS, Ca2+ and products of lipid peroxidation, resulting from H2O2 were significantly reduced by scutellarin. Incubation of cells with H2O2 caused a marked decrease in MMP, which was significantly inhibited by scutellarin. PC12 cells treated with H2O2 underwent apoptotic death as determined by flow cytometric assay. The percentage of this H2O2-induced apoptosis in the cells was decreased in the presence of different concentrations of scutellarin. Scutellarin exhibited significantly higher potency compared to the antioxidant vitamin E. The present findings showed that scutellarin attenuated H2O2-induced cytotoxicity, intracellular accumulation of ROS and Ca2+, lipid peroxidation, and loss of MMP and DNA, which may represent the cellular mechanisms for its neuroprotective action.
9920719
Long‐term and high‐dose piracetam treatment of Alzheimer's disease
Preclinical research suggests that piracetam (a nootropic drug) may improve cognitive functions, but previous studies have failed to demonstrate a clear benefit for the treatment of Alzheimer's disease (AD). We report a 1-year, double-blind, placebo-controlled, parallel-group study with a high dose of piracetam (8 g/d per os) in 33 ambulant patients with early probable AD. Thirty subjects completed the 1-year study. No improvement occurred in either group, but our results support the hypothesis that long-term administration of high doses of piracetam might slow the progression of cognitive deterioration in patients with AD. The most significant differences concerned the recall of pictures series and recent incident and remote memory. The drug was well-tolerated.