Neuroprotective effect of quercetin nanoparticles: A possible prophylactic and therapeutic role in alzheimer’s disease

https://doi.org/10.1016/j.jchemneu.2020.101795Get rights and content

Highlights

  • Alzheimer’s disease (AD) became the most common cause of dementia in elderly.

  • Quercetin is flavonoid with low bioavailability, however its nanoparticles (QNPs) have a better bioavailability.

  • QNPs administration remarkably reduced the neuronal degenerative changes, reduced APs and NFTs formation and increased cellular proliferation.

  • Thus, QNPs blocked the damaging effect of Alcl3 on hippocampal neurons.

  • This is the first study to show a prophylactic and therapeutic effect for QNPs in AD model.

Abstract

Background

Alzheimer’s disease (AD) is the most common cause of dementia in elderly. Quercetin is a well-known flavonoid with low bioavailability. Recently, quercetin nanoparticles (QNPs) has been shown to have a better bioavailability.

Aims

This study aimed to investigate the protective and therapeutic effects of QNPs in Aluminum chloride (AlCl3) induced animal model of AD.

Materials and Methods

AD was induced in rats by oral administration of AlCl3 (100 mg/kg/day) for 42 days. QNPs (30 mg/kg) was given along with AlCl3 in the prophylactic group and following AD induction in the treated group. Hippocampi were harvested for assessments of the structural and ultrastructural changes using histological and histochemical approaches.

Results and Discussion

AD hippocampi showed a prominent structural and ultrastructural disorders both neuronal and extraneuronal. Including neuronal degeneration, formation of APs and NFTs, downregulation of tyrosine hydroxylase (TH), astrogliosis and inhibition of the proliferative activity (all P ≤ 0.05). Electron microscopy showed signs of neuronal degeneration with microglia and astrocyte activation and disruption of myelination and Blood Brain Barrier (BBB). Interestingly, QNPs administration remarkably reduced the neuronal degenerative changes, APs and NFTs formation (all P ≤ 0.05). Furthermore, it showed signs of regeneration (all P ≤ 0.05) and upregulation of TH. The effect was profound in the prophylactic group. Thus, QNPs reduced the damaging effect of AlCl3 on hippocampal neurons at the molecular, cellular and subcellular levels.

Conclusion

For the best of our knowledge this is the first study to show a prophylactic and therapeutic effect for QNPs in AD model. This might open the gate for further research and provide a new line for therapeutic intervention in AD.

Introduction

Alzheimer’s disease (AD) is the most common neurodegenerative disorder impairing memory and cognitive functions (Villemagne et al., 2018). Epidemiological researches have reported that individuals with low schooling levels, brain injury history or sedentary life styles are more likely to develop AD (Reitz et al., 2011). Diabetes mellitus, midlife obesity, hypertension and physical inactivity are the most common predisposing factors for AD are (Hüttenrauch et al., 2016). AD accounts for 80 % of dementia cases worldwide (Loureiro et al., 2017). Neuropathologically, the hallmarks of AD are amyloid plaques (APs) which are spherical extracellular accumulations of Amyloid-β protein and the intracellular neurofibrillary tangles (NFTs) which result from abnormal hyperphosphorylation of cytoskeletal tau protein (Ghoneim et al., 2015). The etiology of AD is still not fully known. However, there are several risk factors triggering the onset of AD, including genetic factors, inflammation, mis-folded proteins accumulation, beta-amyloid (Aβ) protein accumulation, synapse components alteration, and neuronal loss (Ferreiro et al., 2012). Mitochondrial and metabolic dysfunction, apoptosis and excite toxicity have been reported as influential factors in AD (Chen et al., 2014). Defective insulin signaling and decreased glucose utilization may also lead to neuronal dysfunction then death leading to dementia (Gamba et al., 2019). Tyrosine hydroxylase (TH) is the rate-limiting enzyme of catecholamine neurotransmitter biosynthesis. In dopaminergic cells in the brain, tyrosine is converted to l-DOPA by TH. Dopamine can then be converted into other catecholamines, such as noradrenaline and adrenaline. This means that decrease in TH expression is associated with reduced dopamine levels (Santana et al., 2019). Snowden and his coworkers (Snowden et al., 2018) reported that dopamine plays several important roles in the regulating mood and aiding cognitive and motor functions. Impairment TH causes depression and memory loss observed in patients with AD. Several studies reported compromised dopamine neurons in a transgenic model of AD and in AD patients even prior to the Aβ depositions (Nobili et al., 2017; Serra et al., 2018). Dementia is the main diagnostic symptom of AD, it occurs due to sever loss of synapses and neurons that selectively influence particular cell subpopulations in brain areas essential for learning and memory (Trujillo-Estrada et al., 2014).

The hippocampus is one of the earliest brain areas affected by these pathologies (Alawdi et al., 2017). Despite the tremendous efforts to understand the mechanisms and to develop therapeutic agents, there is no effective treatment for AD up till now (Pakdaman et al., 2015). Experimental work showed therapeutic lines such as acetylcholinesterase inhibitors, antioxidants or drugs improving glucose utilization may have a beneficial effect in rats model of AD (Ponce-Lopez et al., 2011).

Aluminum is widely used in the manufactured food, cosmetics (Becaria et al., 2003), food additives, tooth paste and many pharmaceutical products such as antacids (Abbasali et al., 2005). It crosses Blood Brain Barrier (BBB) and persists in the brain for up to five months (Tomljenovic, 2011; Ekong et al., 2017). Rats exposed to low levels of AlCl3 in the drinking water showed higher aluminum level in the brain than the control group (Darbre et al., 2013). Moreover, it induces BBB permeability leading to aluminum and other substances enter the brain (Cabus et al., 2015). Aluminum has neurotoxic effects in animals and humans and is implicated in the pathophysiology of AD, it exacerbates the deposition of beta amyloid (Aβ) with subsequent amyloid plaques (APs) formation (Praticò et al., 2002). The hypothesis that Al induces AD was raised in 1911 (Tomljenovic, 2011), and was approved again 1965 (Lidsky, 2014). Injection of Al salts into rabbits’ brains, showed a correlation between the cognitive deficits and the accumulation of neurofibrillary tangles (NT) (Duwe and Niedzwiecki, 2020). We believe, AlCl3 would create an animal AD model that simulates human AD (Abd El-Aleem et al., 2020). Hence, in this study we have used AlCl3 to induce AD in adult male, Sprague Dawley rats.

Quercetin is a natural flavonoid enriched in vegetables, fruits and other dietary products especially in onions, apples, Ginko Biloba and red wine (Palle and Neerati, 2017). Quercetin is a potent antioxidant, anti-inflammatory and radical-scavenger and potentially have a therapeutic effects in diabetes, infection, cancer, cardiovascular and neurodegenerative diseases (Kong et al., 2016). Interestingly, in vivo and in vitro models of AD showed that quercetin may have a role in AD related disorders. It decreases intracellular tau pathology, extracellular APs and gliosis in the amygdala and the hippocampus of aged triple transgenic AD mice (Sabogal-Guáqueta et al., 2015) and it protects nerve cells and hippocampal cultures against Aβ toxicity in vitro (Choi et al., 2014). However, the exact mechanisms of quercetin ameliorative effects on AD are not fully understood yet (Kong et al., 2016). In spite of these beneficial effects, the pharmacological application of quercetin is limited due to its low oral bioavailability (<2%), low brain permeability and its hydrophobic nature (Kumar et al., 2016). In this study, we have used the nanoparticle form of quercetin which might increase its bioavailability in the brain.

To date, nanoparticles have attracted particular attention in the therapeutics of AD due to their excellent stability, high bioavailability and ready ability of crossing the Blood Brain Barrier (BBB) especially for hydrophobic compounds as quercetin (Sun et al., 2016). In this study, quercetin nanoparticles (QNPs) were prepared by antisolvent precipitation method under sonication (Raval and Patel, 2011) and its bioavailability was enhanced by using Noyes-Whitney equation which state that decreasing the particle sizes would increase the particle surface areas (Palle and Neerati, 2017). This study aims to investigate the protective and the therapeutic effects of QNPs in AlCl3 induced AD and to shed a light on the possible mechanisms of actions.

Section snippets

Animals and chemicals

This study was carried out in the Histology and Cell Biology Department, Faculty of Medicine, Minia University. Adult male, Sprague Dawley rats, 6 per group, weight (150–200 g m) and age (6–8 weeks) matched were housed for seven days prior to the experimentation. All experiments were performed according to regulations under the appropriate animal licenses approved by the animal care committee of Faculty of Medicine-Minia University (Approval No.233: 7/2019), according to the international

Hippocampus mapping and histological features

Normal organization of the hippocampus formation was seen formed of the Cornu Ammonis (CA) and the dentate gyrus (DG) (Fig. 1). The cellular organization in each zone was studied at higher magnification (Fig. 2A–C). In CA1 cells were arranged as 3–4 layers composed of closely packed small pyramidal neurons with vesicular nuclei (Fig. 2A). In contrast, in CA3 cells are loosely packed large pyramidal neurons with vesicular nuclei (Fig. 2B). In DG cells are arranged as dense columns of granular

Discussion

Alzheimer’s disease (AD) became the most common cause of dementia in elderly (Henstridge et al., 2019), it is expected that the number of people with AD will triple by 2050 (Valdez, 2019). Up to date there is no effective therapy for AD (Qin et al., 2019). Quercetin is a multi-tasker agent with antioxidant, anti-inflammatory and neuroprotectant effects (Kong et al., 2016). However, its low bioavailability restricts its usage in clinical applications (Kumar et al., 2016). Nanotechnology

Conclusion

QNPs administration in AD model could preserve structure and function of hippocampal neurons by several mechanisms at cellular, subcellular and molecular levels. It blocked NFTs and APs formation, restored TH activity and enhanced regenerative changes with subsequent neuronal function improvement. Additionally, it affected extra neuronal structures; it regulated astrocyte and microglial activities, preserved myelin sheath and BBB intact. Thus, using nanoparticles of quercetin might provide a

Author contribution statement

Rehab Rifaai: Supervisor, helped in experimental design, data analysis and writing of the manuscript.

Sahar Mokhemer: PhD candidate who performed the experimental lab work and wrote the manuscript.

Entesar Ali: Supervisor, helped in experimental design, data analysis and writing of the manuscript.

Seham Abd El-Aleem: Supervisor, helped in data analysis and revision of the manuscript.

Nashwa El-Tahawy: Supervisor, helped in experimental design, data analysis and writing of the manuscript.

Data availability statement

The main data are included in this manuscript and in its supplementary files. All data are available from the corresponding author on reasonable request

Ethical statement

All experiments were performed according to regulations under the appropriate animal licenses approved by the animal care committee of Faculty of Medicine-Minia University (Approval No.233: 7/2019), according to the international guidelines (Act 1986). Animals were observed daily to assure animal wellbeing.

Declaration of Competing Interest

There is no conflict of interests.

Acknowledgments

We are grateful to Professor Usama Farghaly Aly for help in preparation QNPs. This research did not receive any specific grant from funding agencies.

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