Effect of curcumin-donepezil combination on spatial memory, astrocyte activation, and cholinesterase expressions in brain of scopolamine-treated rats.

BACKGROUND: The study investigated the effect of co-administration of curcumin and donepezil on several markers of cognitive function (such as spatial memory, astrocyte activation, cholinesterase expressions) in the brain cortex and hippocampus of scopolamine-treated rats. METHOD AND RESULTS: For seven consecutive days, a pre-treatment of curcumin (50 mg/kg) and/or donepezil (2.5 mg/kg) was administered. On the seventh day, scopolamine (1 mg/kg) was administered to elicit cognitive impairment, 30 min before memory test was conducted. This was followed by evaluating changes in spatial memory, cholinesterase, and adenosine deaminase (ADA) activities, as well as nitric oxide (NO) level were determined. Additionally, RT-qPCR for glial fibrillary acidic protein (GFAP) and cholinesterase gene expressions was performed in the brain cortex and hippocampus. Also, GFAP immunohistochemistry  of the brain tissues for neuronal injury were performed in the brain cortex and hippocampus. In comparison to the control group, rats given scopolamine had impaired memory, higher levels of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and ADA activities, as well as elevated markers of oxidative stress. In addition to enhanced GFAP immunoreactivity, there was also overexpression of the GFAP and BChE genes in the brain tissues. The combination of curcumin and donepezil was, however, observed to better ameliorate these impairments in comparison to the donepezil-administered rat group. CONCLUSION: Hence, this evidence provides more mechanisms to support the hypothesis that the concurrent administration of curcumin and donepezil mitigates markers of cognitive dysfunction in scopolamine-treated rat model.

Curcumin improves the ability of donepezil to ameliorate memory impairment in Drosophila melanogaster: involvement of cholinergic and cnc/Nrf2-redox systems.

One of the well-established models for examining neurodegeneration and neurotoxicity is the Drosophila melanogaster model of aluminum-induced toxicity. Anti-cholinesterase drugs have been combined with other neuroprotective agents to improve Alzheimer's disease management, but there is not much information on the combination of anti-cholinesterases with dietary polyphenols to combat memory impairment. Here, we assess how curcumin influences some of the critical therapeutic effects of donepezil (a cholinesterase inhibitor) in AlCl3-treated Drosophila melanogaster. Harwich strain flies were exposed to 40 mM AlCl3 - alone or in combination with curcumin (1 mg/g) and/or donepezil (12.5 µg/g and 25 µg/g) - for seven days. The flies' behavioral evaluations (memory index and locomotor performance) were analyzed. Thereafter, the flies were processed into homogenates for the quantification of acetylcholinesterase (AChE), catalase, total thiol, and rate of lipid peroxidation, as well as the mRNA levels of acetylcholinesterase (ACE1) and cnc/NRF2. Results showed that AlCl3-treated flies presented impaired memory and increased activities of acetylcholinesterase and lipid peroxidation, while there were decrease in total thiol levels and catalase activity when compared to the control. Also, the expression of ACE1 was significantly increased while that of cnc/NRF2 was significantly decreased. However, combinations of curcumin and donepezil, especially at lower dose of donepezil, significantly improved the memory index and biochemical parameters compared to donepezil alone. Thus, curcumin plus donepezil offers unique therapeutic effects during memory impairment in the D. melanogaster model of neurotoxicity.

Evaluating Water bitter leaf (Struchium sparganophora) and Scent Leaf (Ocimum gratissimum) extracts as sources of nutraceuticals against manganese-induced toxicity in fruit fly model.

Tropical vegetables remain one of the major sources of functional foods and nutraceuticals, while their constituent phytochemicals, especially alkaloids, have been reported to exhibit neuroprotective properties. Here, the protective effect of alkaloid extracts from Scent leaf (Ocimum gratissimum) and Water bitter leaf (Struchium sparganophora) on manganese (Mn)- induced toxicity in wild type fruit fly (Drosophila melanogaster) model was investigated. Flies were exposed to 30 mM of Mn, the alkaloid extracts (20 and 200 µg/g) and co-treatment of Mn plus extracts, respectively. The survival rate and locomotor performance of the flies were assessed 7 days post-treatment, after which the flies were homogenized and assayed for activities of acetylcholinesterase (AChE), monoamine oxidase (MAO), glutathione-S transferase (GST), catalase, superoxide dismutase SOD), as well as total thiol, reactive oxygen species (ROS) and neural L-DOPA levels. Results showed that the extract significantly reversed Mn-induced reduction in the survival rate and locomotor performance of the flies. Furthermore, both extracts counteracted the Mn-induced elevation in AChE and MAO activities, as well as reduced antioxidant enzyme activities, with a concomitant mitigation of Mn-induced elevated ROS and neural L-DOPA level. The HPLC characterization of the extracts revealed the presence of N-propylamine, Vernomine and Piperidine as predominant in Water bitter leaf extract, while 2, 6-dimethylpyrazine and sesbanimide were found in scent leaf extract. Therefore, the alkaloid extract of these leaves may thus be sources of useful nutraceuticals for the management of pathological conditions associated with manganese toxicity.

Effect of Andrographis paniculata and Phyllanthus amarus leaf extracts on selected biochemical indices in Drosophila melanogaster model of neurotoxicity.

Aluminum (Al)-induced toxicity in fruit fly (Drosophila melanogaster) is one of the established models for studying neurotoxicity and neurodegenerative diseases. Alkaloid phytochemicals have been reported to exhibit neuroprotective effects. Therefore, the aim of this study is to determine the effect of alkaloid extracts of Andrographis paniculata and Phyllanthus amarus leaves on Al-induced toxicity in wild type Drosophila melanogaster. The flies were exposed to diets containing 40 mM AlCl3, and the alkaloid extracts (0.1 and 1 mg/ml). Thereafter, the flies were assessed for learning and memory, as well as locomotor performance 14 days post-treatment. This was followed by homogenizing the flies and the homogenates were assayed for acetylcholinesterase, monoamine oxidase and catalase activities, as well as the malondialdehyde content. The results showed that the alkaloid extracts of both leaves could ameliorate the aluminum-induced behavioral and biochemical impairments in the flies. The HPLC analysis of the alkaloid contents revealed that there is an abundance of Amaryllidaceae alkaloids, caffeine and carpaine. Thus, alkaloid extracts from these leaves could serve as promising therapeutic candidates for the management of neurodegenerative disease.

Characterization and neuroprotective properties of alkaloid extract of Vernonia amygdalina Delile in experimental models of Alzheimer's disease.

This study emphasized on the neuroprotective properties of bitter leaf alkaloid-rich extract (BLAE) using transgenic fruit fly (Drosophila melanogaster [D. melanogaster]) model and scopolamine-induced amnesia rats. In vitro antioxidant properties and modulatory effects on key neuronal enzymes were carried out. Thereafter, fruit flies expressing human amyloid precursor protein (hAPP) and BACE-1 genes were treated with BLAE for 7 d to determine survival rate, BACE-1, acetylthiocholine (AChE), glutathione-S-transferase (GST), and catalase activities. Also, the aftermath of the BLAE on the neuronal activities of AChE, butyrylcholine (BChE), monoamine oxidase (MAO), angiotensin-I converting enzyme (ACE), ATP diphosphohydrolase (ATPdase), and ADPdase, catalase, superoxide dismutase (SOD), plus TBARS, and nitric oxide (NO) content in rats treated with scopolamine (1 mg/kg. bwt. i.p.) was evaluated. In addition, the alkaloid characterization for constituent BLAE was determined. The outcomes proved that BLAE displayed antioxidant properties and inhibit activities of AChE, BChE, MAO, ACE, ATPdase, and ADPdase in vitro. Furthermore, transgenic flies treated with the BLAE exhibited significant levels of amelioration on survival rate and activities of BACE-1, AChE, GST, and catalase. In scopolamine-treated rats, AChE, BChE, MAO and NTPdases activities, and antioxidant status were upturned in rats pretreated with BLAE. This study disclosed the neuroprotective property of BLAE, which could be related to its alkaloid constituent, thereby making it a good candidate to explore as curative nutraceutical agent for cognitive impairments and affiliated diseases such as AD.

Does caffeine influence the anticholinesterase and antioxidant properties of donepezil? Evidence from in vitro and in vivo studies.

Caffeine is adjudged world's most consumed pharmacologically active food component. With reports of the potential cognitive enhancing properties of caffeine, we sought to investigate if caffeine can influence the anticholinesterase and antioxidant properties of donepezil-a selective acetylcholinesterase (AChE) inhibitor used in the management of Alzheimer's disease (AD). In vitro, we investigated the effect of donepezil (DON), caffeine (CAF) and their various combinations on the activity of AChE in rat brain homogenate, as well as determined their antioxidant properties. In vivo, two rat groups were administered single oral dose of DON (5 mg/kg) and CAF (5 mg/kg) separately, while three groups, each received 5 mg/kg DON plus either 5, 50 or 100 mg/kg CAF for three hours, after which the rats were sacrificed and brain isolated. Results show that CAF concentration dependently and synergistically increased the anticholinesterase properties of DON in vitro. Also, CAF produced a significant influence on investigated in vitro antioxidant properties of DON. Furthermore, rats administered 5 mg/kg CAF and DON produced no significant difference in AChE activity compared to rats administered DON alone. However, co-administration of either 50 or 100 mg/kg CAF with DON lead to higher AChE activity compared to both control and DON groups. In addition, DON, CAF and their various combinations augmented brain antioxidant status in treated rats. We conclude that while low caffeine consumption may improve the antioxidant properties of donepezil without having a significant influence on its anticholinesterase effect, moderate-high caffeine consumption could also improve the antioxidant properties of donepezil but reduce its anticholinesterase effect; nevertheless, a comprehensive clinical trial is essential to fully explore these possibilities in human AD condition.

In the quest for the ideal sweetener: Aspartame exacerbates selected biomarkers in the fruit fly (Drosophila melanogaster) model of Alzheimer's disease more than sucrose.

This study evaluated the effect of dietary inclusions of aspartame and sucrose on some selected behavioral and biochemical indices linked with Alzheimer's disease in a transgenic fruit fly (Drosophila melanogaster) model expressing human amyloid precursor protein and secretase. Flies were raised on a diet supplemented with sucrose and aspartame for 14 days. Thereafter, the flies were assessed for their survival rate, learning and memory, as well as locomotor performance, 14 days post-treatment. This was followed by homogenising the fly heads, and the homogenates were assayed for acetylcholinesterase and monoamine oxidase activities, as well as levels of lipid peroxidation, reactive oxygen species, and total thiol. The results showed aspartame at all levels of dietary intake and a high proportion of sucrose significantly aggravated the mortality rate, locomotor deficiency, and impaired biomarkers of oxidative stress and antioxidant status in the transgenic flies, while no significant effect was found on acetylcholinesterase activity or memory function. These findings therefore suggest that while low dietary inclusions of sucrose are tolerable under AD-like phenotypes in the flies, high inclusions of sucrose and all proportions of aspartame tested aggravated mortality rate, locomotion and oxidative stress in the flies.

Phytochemicals from African eggplants (Solanum macrocarpon L) and Black nightshade (Solanum nigrum L) leaves as acetylcholinesterase inhibitors: an in-silico study.

Acetylcholinesterase inhibitors (AChEIs) like donepezil are commonly used to treat Alzheimer's disease. AChEIs have also been considered for other therapeutic uses, such as anti-inflammatory neuroprotective agents. Consequently, the use of natural plant products as potential AChEIs can have therapeutic benefits. We previously reported the anticholinesterase properties of the phenolics and alkaloids found in the leaf extracts of two tropical plants with nutritional and ethnobotanical importance-African eggplant (Solanum macrocarpon L) and Black nightshade (Solanum nigrum L). Here, we tested the ability of both extracts to inhibit human erythrocyte AChE (an indirect mediator of pro-inflammatory cytokines production via acetylcholine degradation). We further used molecular docking and MD simulation to identify the potential molecular mechanism(s) of phenolic and alkaloid compounds as human AChEIs. Special focus was given to compounds containing the benzyl group that can establish stacking interactions similar to donepezil (a standard AChEI). Flavone-luteolin rutinosides (LR) were identified as single-binding or dual-binding AChEIs; specifically, we attributed the dual-binding LR4 and LR5 to their linked hexose moiety. This characteristic allows the dual binders to occupy the catalytic triads and the peripheral anionic subsite, while exploring the catalytic gorge. We further delineated the inhibition of human erythrocyte AChE, as the flavone common to both plant extracts-luteolin rutinosides-had positive in silico interactions with AChE. These findings suggest that phytochemicals from S. macrocarpon and S. nigrum with dual binding properties can be potential AChE inhibitors. In fact, compounds such as LR4 and LR5 should be further investigated as potential inhibitors of human AChE and may represent important natural alternatives to donepezil.Communicated by Ramaswamy H. Sarma.

Can gallic acid potentiate the antihyperglycemic effect of acarbose and metformin? Evidence from streptozotocin-induced diabetic rat model.

This study investigated the influence of dietary phenolic acid- Gallic acid (GA) on the antihyperglycemic properties of acarbose (ACA) and metformin (MET). Streptozotocin-induced diabetic rats were treated (p.o) with ACA, MET, GA and their combinations for 14 days. The effects of the treatments on blood glucose and insulin levels, pancreas α-amylase and intestinal α-glucosidase activities, as well as thiobarbituric acid reactive species (TBARS), thiol and reactive oxygen species (ROS) levels, including antioxidant enzyme activities were investigated. A significant increase in blood glucose, insulin, ROS and TBARS levels, and impaired antioxidant status, as well as elevation in the activities of α-amylase and α-glucosidase observed in diabetic rats were ameliorated in the treatment groups. Hpwever, GA had varying effects on the antidiabetic properties of the drugs. Nevertheless, GA showed more potentiating effects on the antidiabetic effect of MET and these effects were better observed at the lower dose of GA.

Citrus spp. essential oils improve behavioral pattern, repressed cholinesterases and monoamine oxidase activities, and production of reactive species in fruit fly (Drosophila melanogaster) model of Alzheimer's Disease.

Much emphasis has been placed on the biological activities of citrus peel's essential oils (CPEOs) against human ailments. This study investigated the effect of Citrus limon and Citrus reticulata peel's essential oils (EOs) on behavioral and neurochemical imbalance in transgenic and Harwish (Wild) fruit flies. Flies were divided into seven groups comprising of the control and those that were fed with 0.1, 0.5, and 1.0 µg/ml of the dietary inclusions of study CPEOs for 7 days. Thereafter, behavioral profile was examined using lethality response and negative geotaxis assays. Effect of the EOs on cholinesterase and monoamine oxidase (MAO) activities, and antioxidative parameters were determined. The result showed a significant improvement of behavioral pattern and biochemical parameters of the flies fed with studied CPEOs inclusive diets. Conclusively, both EOs exert neuroprotective capability by reducing cholinesterases and monoamine activities, and also prevent oxidative stress, which are implicated in neuronal dysfunction in humans. PRACTICAL APPLICATIONS: With the growing increase in the search for safer alternatives, having no side effects, for the management of neurodegenerative diseases, a large proportion of the populace is beginning to find solace in the use of natural products. Also, the wide array of similarities between the humans and the dipteran insects, fruit flies is a perfect organism for the study of neurodegenerative diseases. Therefore, this study presents the neuroprotective potentials of lemon and tangerine peels-derived EOs, and the possibility of their exploration as neuroactive agents and alternative in the management of Alzheimer's disease (AD).

Influence of gallic and tannic acid on therapeutic properties of acarbose in vitro and in vivo in Drosophila melanogaster.

BACKGROUND: In this study, gallic acid (GA) and its polymeric form-tannic acid (TA) which are two phenolic acids found abundantly distributed in plant food sources were investigated for their influence on therapeutic properties of acarbose (AC) in vitro and in vivo in Drosophila melanogaster. METHODS: Combinations of AC and GA or TA were assessed for their alpha-glucosidase and alpha-amylase inhibitory effects as markers of anti-hyperglycemic properties, as well as their free radicals scavenging, Fe2+ chelating and malondialdehyde (MDA) inhibitory effects (in vitro). Furthermore, wild type D. melanogaster cultures were raised on diets containing AC, GA, TA and their various combinations for seven days. Thereafter, flies were homogenized and glucose concentrations, alpha-glucosidase and alpha-amylase activities, as well as reactive oxygen species (ROS) and total thiol levels were determined. RESULTS: The results showed that GA and TA up to 5 mg/ml significantly (p < 0.05) increased the enzymes' inhibitory effects and antioxidant properties of AC in vitro. Also, there was significant reduction in glucose concentration, enzyme activities and ROS level in D. melanogaster fed diets supplemented with phenolic acids and acarbose. CONCLUSIONS: These bioactive compounds-drug interactions provide useful information on improving the therapeutic properties of acarbose especially in its use as an antidiabetic drug.

Effect of Alkaloid Extract from African Jointfir (Gnetum africanum) Leaves on Manganese-Induced Toxicity in Drosophila melanogaster.

Metal-induced toxicity in fruit fly (Drosophila melanogaster) is one of the established models for studying neurotoxicity and neurodegenerative diseases. Phytochemicals, especially alkaloids, have been reported to exhibit neuroprotection. Here, we assessed the protective effect of alkaloid extract from African Jointfir (Gnetum africanum) leaf on manganese- (Mn-) induced toxicity in wild type fruit fly. Flies were exposed to 10 mM Mn, the alkaloid extract and cotreatment of Mn plus extract, respectively. The survival rate and locomotor performance of the flies were assessed 5 days posttreatment, at which point the flies were homogenized and assayed for acetylcholinesterase (AChE) activity, nitric oxide (NO), and reactive oxygen species (ROS) levels. Results showed that the extract significantly reverted Mn-induced reduction in the survival rate and locomotor performance of the flies. Furthermore, the extract counteracted the Mn-induced elevation in AChE activity, NO, and ROS levels. The alkaloid extract of the African Jointfir leaf may hence be a source of useful phytochemicals for the development of novel therapies for the management of neurodegeneration.

Influence of gallic acid on α-amylase and α-glucosidase inhibitory properties of acarbose.

Acarbose is an antidiabetic drug which acts by inhibiting α-amylase and α-glucosidase activities but with deleterious side effects. Gallic acid (GA) is a phenolic acid that is widespread in plant foods. We therefore investigated the influence of GA on α-amylase and α-glucosidase inhibitory properties of acarbose (in vitro). Aqueous solutions of acarbose and GA were prepared to a final concentration of 25µM each. Thereafter, mixtures of the samples (50% acarbose + 50% GA; 75% acarbose+25% GA; and 25% acarbose+75% GA) were prepared. The results revealed that the combination of 50% acarbose and 50% GA showed the highest α-glucosidase inhibitory effect, while 75% acarbose+25% GA showed the highest α-amylase inhibitory effect. Furthermore, all the samples caused the inhibition of Fe2+-induced lipid peroxidation (in vitro) in rat pancreatic tissue homogenate, with the combination of 50% acarbose and 50% GA causing the highest inhibition. All the samples also showed antioxidant properties (reducing property, 2,2'-azino-bis (-3-ethylbenzthiazoline-6-sulphonate [ABTS*] and 1,1-diphenyl-2-picrylhydrazyl [DPPH] free radicals scavenging abilities, and Fe2+ chelating ability). Therefore, combinations of GA with acarbose could be employed as antidiabetic therapy, with a possible reduction of side effects of acarbose; nevertheless, the combination of 50% acarbose and 50% GA seems the best.

Polyphenolic compositions and in vitro angiotensin-I-converting enzyme inhibitory properties of common green leafy vegetables: A comparative study.

This study compared the phenolic compositions of common green leafy vegetable extracts from Vernonia amygdalina (VA), Telfairia occidentalis (TO), Talinium triangulare (TT), and Amaranthus hybridus (AH) and their effects on the angiotensin-I-converting enzyme (ACE) and cisplatin-induced malonylaldehyde (MDA) production in an isolated rat kidney homogenate. HPLC confirmed the presence of phenolic compounds in the extracts. Furthermore, all extracts inhibited ACE activity dosedependently; however, the extract from VA exhibited the highest ACE activity while TT exhibited the least. Incubation of the kidney homogenate with 1mM cisplatin caused an increase in MDA production; however, all the extracts inhibited the level of MDA produced. Nevertheless, VA extract exhibited the highest inhibition. These activities of the vegetable extracts could be attributed to their phenolic compositions and may suggest some possible mechanism of the actions. However, VA appeared to be the most potent among the vegetables tested.