Neuroprotective Effects of Agri-Food By-Products Rich in Phenolic Compounds

Neurodegenerative diseases are known for their wide range of harmful conditions related to progressive cell damage, nervous system connections and neuronal death. These pathologies promote the loss of essential motor and cognitive functions, such as mobility, learning and sensation. Neurodegeneration affects millions of people worldwide, and no integral cure has been created yet. Here, bioactive compounds have been proven to exert numerous beneficial effects due to their remarkable bioactivity, so they could be considered as great options for the development of new neuroprotective strategies. Phenolic bioactives have been reported to be found in edible part of plants; however, over the last years, a large amount of research has focused on the phenolic richness that plant by-products possess, which sometimes even exceeds the content in the pulp. Thus, their possible application as an emergent neuroprotective technique could also be considered as an optimal strategy to revalorize these agricultural residues (those originated from plant processing). This review aims to summarize main triggers of neurodegeneration, revise the state of the art in plant extracts and their role in avoiding neurodegeneration and discuss how their main phenolic compounds could exert their neuroprotective effects. For this purpose, a diverse search of studies has been conducted, gathering a large number of papers where by-products were used as strong sources of phenolic compounds for their neuroprotective properties. Finally, although a lack of investigation is quite remarkable and greatly limits the use of these compounds, phenolics remain attractive for research into new multifactorial anti-neurodegenerative nutraceuticals.


Introduction
The neurodegeneration process is based on the continuous dysfunction of nerve structures and neuronal loss, progressively causing a detrimental effect on cognitive and motor-related abilities, such as memory, learning, decision making, balance, movement, talking, breathing and heart function [1]. Among age-related diseases, chronic neurodegeneration prevalence has increasingly grown over the last decades, being one of the most current urgent health concerns in society [2].
The most common neurodegenerative disorders are Alzheimer's disease (AD) and Parkinson's disease (PD). In AD, the inhibition of correct neuronal function hinders one's communication, which finally leads to memory loss, cognitive decline and dementia. This condition affects about 1 in 9 people (10.7%) aged 65 and older, and about 11 in 10000 people (0.11%) under 65 in the U. S. [3]. It was estimated that around 58.66 million people suffered from AD in 2020 worldwide, a number expected to triplicate by 2050 [4]. PD symptoms are more related to motor function diminishing, such as resting tremor, postural imbalance, bradykinesia and muscular rigidity [5,6]. In 2019, it was estimated that more than 10 million people worldwide lived with this disease [7].

Inclusion and Exclusion Criteria
A total of 2599 publications resulted from the core search. Only articles from 2018 to 2022 written in English language were firstly identified. Then, the result was refined using specific citation topics and research categories (phytochemicals, neurodegenerative diseases, neuroscience and food science and tech., cell biology, chemistry, biochemistry, nutrition and dietetics and plant sciences). In terms of the Open Access Status, papers categorized as "Bronze" were rejected since the licensing for these articles was either unclear or nonexistent. Additionally, "Green Submitted/Accepted" statuses were rejected, since the then readable version was not definitive. After this, the title and abstract screening took place, taking into account the citation score. Different issues were considered for rejection: isolated compounds, extracts from other species or by-products (Fungi kingdom, juices and infusions, etc.), other biomolecules (peptides, fatty acids, sugars), focus on other close pathologies (obesity, diabetes) and too generalist or redundant information. Resulting articles were revised, and those that did not specifically cover neuroprotection or that did not provide particularly relevant information due to the nature of the study were excluded. 70 articles were obtained and used for the investigation. Among selected articles there were in vitro and in vivo studies, so they were gathered into different tables for a better understanding. In addition, in silico and ex vivo studies were also considered.

Inclusion and Exclusion Criteria
A total of 2599 publications resulted from the core search. Only articles from 2018 to 2022 written in English language were firstly identified. Then, the result was refined using specific citation topics and research categories (phytochemicals, neurodegenerative diseases, neuroscience and food science and tech., cell biology, chemistry, biochemistry, nutrition and dietetics and plant sciences). In terms of the Open Access Status, papers categorized as "Bronze" were rejected since the licensing for these articles was either unclear or nonexistent. Additionally, "Green Submitted/Accepted" statuses were rejected, since the then readable version was not definitive. After this, the title and abstract screening took place, taking into account the citation score. Different issues were considered for rejection: isolated compounds, extracts from other species or by-products (Fungi kingdom, juices and infusions, etc.), other biomolecules (peptides, fatty acids, sugars), focus on other close pathologies (obesity, diabetes) and too generalist or redundant information. Resulting articles were revised, and those that did not specifically cover neuroprotection or that did not provide particularly relevant information due to the nature of the study were excluded. 70 articles were obtained and used for the investigation. Among selected articles there were in vitro and in vivo studies, so they were gathered into different tables for a better understanding. In addition, in silico and ex vivo studies were also considered. Several articles which were included did not fulfil the established criteria according to publication date; regardless, they were included for their relevance to the study.
For in vivo articles, a PICO framework (Patient or Population, Intervention, Comparison and Outcome) was established. Humans and all types of animals of any gender and age were considered valid for the review. In terms of diseases, patients who suffered from AD or PD, or from any pathology or symptom related to neurodegeneration, such as induced oxidative stress or neuroinflammation, were preferred. On the other hand, drug treatment

Protein Misfolding, Aggregation and Deposition
Protein abnormal interactions such as misfolding, aggregation and deposition are considered common pathological hallmarks of neurodegenerative disorders.
Amyloid-β (Aβ) peptides perform different functions in the CNS, and is progressively accumulated in the extracellular space of the brain as a consequence of the sequential cleavage of amyloid precursor proteins (APP) by β-secretase activity [27]. The overproduction of insoluble Aβ, common in AD, forms toxic oligomers which eventually accumulate in senile plaques, which are deposited throughout the brain [2,28]. This condition is usually accompanied by damage to the tau proteins, peptides responsible for the structural support of microtubules in neurons. In this sense, Aβ plaques cause a hyperphosphorylation of tau proteins, which translates into the formation of new neurotoxic tau tangles, known as neurofibrillary tangles (NFT) [27]. This event causes synaptic impairment, induced inflammation through microglia activation and, ultimately, cell death [9,29]. Moreover, the ubiquitin-proteasome system (UPS), the main intracellular proteolytic system which regulates the production and degradation of proteins, is also affected. Its dysfunction, caused by senile plaques and NFTs, eventually leads to further aggregation [26]. Ubiquitinated proteins have previously been detected in senile plaques and NFTs, demonstrating the interrelationship between UPS dysfunction, protein aggregation and deposition inside and outside the neuron [28].
Amyloid plaques are capable of passing to the brain through the blood, damaging neurons and activating different nervous cells such as microglia cells and astrocytes. These events are closely related to other phenomena such as high free radical production, excitotoxicity, increase in neuronal apoptosis and promotion of the synthesis of proinflammatory molecules, which leads to neuroinflammation [2,30,31].
Notwithstanding, new studies are proving that, in contrast to the extracellular deposition of Aβ as AD main initiator, these plaques are indeed the result of intraneuronal accumulation of Aβ inside brain cell lysosomes [32]. Due to the novelty of the finding, the

Protein Misfolding, Aggregation and Deposition
Protein abnormal interactions such as misfolding, aggregation and deposition are considered common pathological hallmarks of neurodegenerative disorders.
Amyloid-β (Aβ) peptides perform different functions in the CNS, and is progressively accumulated in the extracellular space of the brain as a consequence of the sequential cleavage of amyloid precursor proteins (APP) by β-secretase activity [27]. The overproduction of insoluble Aβ, common in AD, forms toxic oligomers which eventually accumulate in senile plaques, which are deposited throughout the brain [2,28]. This condition is usually accompanied by damage to the tau proteins, peptides responsible for the structural support of microtubules in neurons. In this sense, Aβ plaques cause a hyperphosphorylation of tau proteins, which translates into the formation of new neurotoxic tau tangles, known as neurofibrillary tangles (NFT) [27]. This event causes synaptic impairment, induced inflammation through microglia activation and, ultimately, cell death [9,29]. Moreover, the ubiquitin-proteasome system (UPS), the main intracellular proteolytic system which regulates the production and degradation of proteins, is also affected. Its dysfunction, caused by senile plaques and NFTs, eventually leads to further aggregation [26]. Ubiquitinated proteins have previously been detected in senile plaques and NFTs, demonstrating the interrelationship between UPS dysfunction, protein aggregation and deposition inside and outside the neuron [28].
Amyloid plaques are capable of passing to the brain through the blood, damaging neurons and activating different nervous cells such as microglia cells and astrocytes. These events are closely related to other phenomena such as high free radical production, excitotoxicity, increase in neuronal apoptosis and promotion of the synthesis of proinflammatory molecules, which leads to neuroinflammation [2,30,31].
Notwithstanding, new studies are proving that, in contrast to the extracellular deposition of Aβ as AD main initiator, these plaques are indeed the result of intraneuronal accumulation of Aβ inside brain cell lysosomes [32]. Due to the novelty of the finding, the Nutrients 2023, 15, 449 6 of 33 information is scarce, but sets a new point of view for the study of AD neurodegeneration and its mechanisms.
In PD, dementia can be promoted by the spread of alpha-synuclein (αS), a protein that normally originates in the enteric nervous system, with subsequent spread to the rest of the CNS [33]. Physiological functions of this peptide include synaptic vesicle recycling, neurotransmission and synaptic plasticity, among others. Although it is usually soluble, αS can accumulate and form insoluble fibrils that, associated with molecules such as ubiquitin, form clusters known as Lewis bodies [33]. These formations can cause synaptic impairments, mitochondrial dysfunctions, membrane disturbances and neuroinflammation through microglia activation. The spread of αS throughout the system allows it to interact with biological and pathological proteins such as Aβ and tau, creating a vicious cycle that enhances neurodegeneration [34].

Oxidative Stress
Oxidative stress is a deleterious condition caused by imbalances between endogenous pro-and antioxidant species. It is one of the most multifaceted disease triggers in the human body and one of the most important biological mechanisms associated with neurodegeneration in AD and PD [35,36].
Reactive oxygen species (ROS) are chemically reactive molecules produced as byproducts derived from the mitochondrial aerobic respiratory chain [23]. In balanced concentrations, they exert essential physiological functions, especially as messenger intermediates in cell signaling processes such as inflammation, cell survival, immune response and synaptic plasticity [37,38]. Nevertheless, an overproduction of ROS can overcome the endogenous antioxidant defense system, composed mainly of the superoxide dismutase (SOD) and the glutathione (GSH) systems. This leads to the deleterious condition of oxidative stress, which affects cell functions and damages different biomolecules through protein oxidation, DNA degradation and lipid peroxidation (LPO) [13,38]. The CNS is particularly vulnerable to oxidative stress due to the high levels of oxygen consumption it requires for a proper functioning, the weakness of its antioxidant system, a limited cellular regeneration capacity and a high content of polyunsaturated fatty acids in neuronal membranes, which are prone to oxidation [36][37][38].
There are different sources of ROS overproduction. Mitochondrion malfunctioning during ATP molecule supply, normally caused by aging or other stressors, is thought to be the main reason. This unbalances the redox system and causes oxidative damage throughout the system [36]. In turn, mitochondrial DNA is also quite sensitive to oxidation, which provokes an even bigger ROS production and leads to a new cyclic problem [39].
Inflammatory responses triggered by brain protein aggregation are considered another important intracellular ROS source in the CNS [39]. In AD, ROS presence oxidizes biomolecules such as proteins, DNA and carbohydrates, which are gradually accumulated in the cell over the years. Neurons, in an attempt to prolong cell life, promote Aβ secretion to sequester ROS, causing their oligomerization and aggregation [40]. Aβ, in turn, is directly involved in ROS formation through peptidyl radicals, metal association or indirect activation of microglia [37]. Therefore, high levels of ROS can be related to neurodegeneration, but they are not considered to be a direct initiator of neuropathies [41][42][43].
In PD, the loss of dopaminergic neurons is closely related to the presence of large amounts of ROS and free radicals through neuroinflammation, dopamine degradation, mitochondrial dysfunction, aging and GSH depletion, among others [37]. Dopamine synthesis is closely related to intracellular oxidation, which makes dopaminergic neurons in substantia nigra especially sensitive to oxidative stress [44].

Neuroinflammation
Neuroinflammation is a pathological process characterized by chronic inflammatory reactions in the CNS which highly contributes to the development of a neurodegenerative state in the brain, mainly AD and PD [26].
Inflammation is a biological mechanism used by an organism against the appearance of an injury or infection through the physiological production of pro-inflammatory markers. However, if this impairment is not corrected in a timely manner, several chronic conditions may be promoted, causing negative health effects [45]. The immune system plays an essential role in this. Microglia represent the most present innate immune cells in the brain and influence other nervous cells such as astrocytes and neurons. Under normal conditions, microglia are deactivated and exert anti-inflammatory responses, but when invaded by pathogens or tissue injury, they are activated. Here, pro-inflammatory responses are promoted, initiating tissue repair cascades that are self-controlled once the repairment has been performed. However, persistent stimulus of altered genes or endogenous factors, such as protein aggregates or ROS overproduction, can confuse the immune system causing uncontrolled inflammation, which generates neurotoxic mediators such as cytokines and interleukins (ILs) that enhance neurodegeneration [44,46].
In AD, the inflammatory response is primarily initiated by senile plaques and NFTs, which trigger inflammatory activation of the closest glial cells through different cascades. The main signaling pathways are the toll-like receptor-4 (TLR-4) pathway, transmembrane proteins expressed in neurons and glial cells involved in innate immune responses via activation of microglia and regulation of pro-inflammatory molecule release, and the mitogen-activated protein kinase (MAPK) pathway [47,48]. Once those microglia and astrocytes near the senile plaques are activated, the release of inflammatory mediators occurs, promoting the secretion of cytokines, chemokines, nuclear factor-Kb proteins, tumor necrosis factor-α and -β, enzymes that metabolize arachidonic acid, such as 5-lipoxygenase (5-LOX) and cyclooxygenase 2 (COX-2), and other compounds such as ROS and glutamate, whose excessive formation induces chronic neuroinflammation and cell death [49]. COX-2 upregulates glial cell activity and synthetizes important pro-inflammatory mediators such as prostaglandins, and LOX-5 catalyzes the formation of leukotrienes, lipid mediators of inflammation. The presence of high levels of lipoxygenase has been related to Aβ overgeneration and tau hyperphosphorylation [9,50].
In PD, the formation of Lewis bodies also promotes the activation of non-neuronal cells, microglia and astrocytes. In addition, αS itself has direct proinflammatory activity. Oligomeric depositions of this peptide are phagocytosed by microglia, followed by their activation, and ROS and NO production, mostly produced by NADPH oxidase in activated microglia [44,51]. Proinflammatory cascades then take place in the same way as explained above.

Impaired Biodynamics and Mitochondrial Dysfunctions
Mitochondria are one of the most important organelles in cells. Their essential role in body homeostasis is related to their capacity to produce energy through ATP formation and regulate calcium homeostasis, their redox signaling and cellular senescence [38]. Due to the high energy demands of the brain, the functions of the mitochondria are especially crucial in neurons for proper synaptic connections and Ca 2+ storage [52]. Since it is the only organelle with its own genome (mtDNA), mitochondrion is quite sensitive to stressors such as aging and protein aggregation, which promotes mtDNA mutation and a decrease in ATP production [53]. Dysfunctional energy production also affects to the ionic gradients across membranes, which ultimately leads to mitochondrial membrane depolarization and permeabilization (MMP), causing devastating neurological effects [38,39].
Mitochondria control programmed cell death, as mitochondrion can induce apoptosis via caspase mechanisms through different proteins [53]. Apoptosis is normally triggered intrinsically by the mitochondria. Once the apoptotic signal pathway is activated, mitochondrial membrane permeability increases and results in the formation of the mitochondrial permeability transition pore. It allows the release into the cytoplasm of pro-apoptotic cytochrome c or caspase activators. Finally, after translocation to the nucleus, the DNA is fragmented, leading to cell death [9,54]. Mitophagy is the apoptotic mechanism of mitochondria to remove damaged areas through self-phagocytosis, and is regulated by the PI3K/Akt/mTOR/AMPK signaling pathway [55]. The process is promoted by AMPK activating ULK-1; however, mTOR can phosphorylate ULK proteins and prevent autophagy. In addition, Akt is able to regulate apoptosis by directly influencing the release of cytochromes and the consequent activation of caspases. AMPK also works as a cellular energy sensor as it is activated by a decrease in ATP; then, autophagy is enhanced by AMPK upregulation or mTOR downregulation [55,56].
Dysfunctional and unbalanced apoptosis sets a starting point for the development of neurodegenerative diseases [53,57]. In AD, Aβ plaques tend to accumulate and aggregate in the mitochondria, interacting with them and blocking electron transport, compromising ATP production and synapsis, in addition to causing cytochrome C release, all proapoptotic signals [38]. In PD, the accumulation of αS oligomers causes MMP, closely related to the enhancement of ROS and the hyperactivation of glial cells. As a result, the neuronal nitric oxide synthase (nNOS), responsible for the production of NO messengers, is wildly activated, worsening pathological conditions. Mitochondrial impairments also affect skeletal muscles and platelets cells, which explains the progressive motor impairment suffered during PD [53]. All of the above has been considered the primary mechanism of dopaminergic neurodegeneration in PD [58].
On the other hand, bound to the outer membranes of mitochondria, there are monoamine oxidase enzymes (MAO), responsible for neurotransmitter inactivation. In fact, MAO-A and MAO-B can degrade dopamine, which exerts vital roles in the regulation of movement. Therefore, their overactivation can lead to dangerous levels of dopamine depletion, promoting mobility impairments and PD [59].

Excitotoxicity-Glutamatergic and Cholinergic Neurotransmissions
The balance between excitatory and inhibitory neuronal synapsis is important for the healthy state of the CNS. These excitatory signals are mainly regulated by glutamate, the predominant neurotransmitter [60]. Glutamatergic neurons are involved in the proper functioning of several cognitive, motor, sensory and autonomic activities, so their levels must be maintained at a physiological level to avoid neuronal impairments. Acetylcholine (ACh) is another important neurotransmitter involved in nerve-impulse transmission between cholinergic neurons. It is also involved in a large number of physiological processes, such as attention, learning, memory, stress response, wakefulness and sleep and sensory information [61]. Therefore, an overabundance of excitatory neurotransmitter levels within the synapse, known as excitotoxicity, has damaging effects on the CNS, as does a depletion of them [62,63].
In average neuronal activity, glutamate is released from its vesicles into the synaptic cleft. Then, it diffuses across and binds to ionotropic glutamate receptors, such as the N-methyl D-aspartate receptor (NMDAR), responsible for forming ion channels that cause cation influx and enable basis neuronal communication [64]. An altered glutamate homeostasis can be achieved through different mechanisms. In AD, senile plaques and/or oxidative stress are able to disturb glutamatergic networks and lead to glutamate overproduction and NMDAR overactivation [65]. This provokes malfunctions such as the excessive intracellular calcium passage, the enhancement of nNOS activity and the promotion of mitochondrial damage with inhibition of the respiratory chain, ATP depletion and necrotic cell death. Furthermore, intramitochondrial calcium is generated, enhancing MMP and leading to caspase cascade activation and apoptosis [66].
In PD, alterations in glutamatergic neurotransmission have been shown to be important triggers of the disease, since glutamate intervenes in several fronto-basal circuits involved in the modulation of voluntary movements [60]. Consequently, dopaminergic neurons begin to degenerate, leading to excessive promotion of glutamatergic concentra-tion and, as abovementioned, to overactivation of NMDAR. These excitotoxic effects are accompanied by increased ROS levels, which increase neuroinflammation and exacerbate damage to nigral dopaminergic neurons in a vicious cyclical problem [66]. Nowadays, it cannot be determined whether inflammation is a cause or a consequence of glutamate unbalance [60].
On the other hand, the loss of cholinergic neurons is also a common hallmark in neurodegenerative diseases such as AD [67]. A consistent ACh depletion and decline in choline acetyltransferase activity are considered to play a key role in learning and memory deterioration in AD patients [68]. This depletion is caused by overactivation of acetylcholinesterase (AChE), a key enzyme in the cholinergic synapsis. Under normal conditions, AChE is responsible for ACh regulation, turning it into choline and acetate molecules. However, overexposure to stressors such as ROS may motivate AChE activity, deplete ACh concentrations to neurodegenerative levels and promote neurological disorders such as dementia [69]. In addition, the strong connection established between AChE and Aβ proteins has been demonstrated; AChE binds to amyloid proteins through its peripheral anionic site, which promotes protein misfolding, fibril formation and neurotoxicity enhancement [50].

Phenolic Compounds from Fruit and Vegetable By-Products: Neuroprotective Mechanisms for Potential Biological Targets
Despite advances in addressing the main mechanisms of neurodegeneration, there is still a serious lack of resources and treatments, and current therapies take little account of the multifactorial behavior of neuronal disorders, causing low efficacy and multiple side effects [70]. Modern medicine works on developing new natural therapeutic agents due to their extremely vast array of biological activities and health benefits [71]. In this sense, phenolic compounds from plants have been demonstrated to possess great bioactivity, either as free radical scavengers, hydrogen atom donors or metal ion chelators [23].
Regarding neuroprotection, phenolic compounds show outstanding bioactivity in numerous pathogenic processes related to aging and neurodegeneration: downregulation of oxidative stress and pro-inflammatory cytokine expression, apoptosis regulation, and activation of proteolysis pathways such as the UPS for preventing protein aggregation, among others [9,12,54]. Flavonoids have been proved to prevent ROS formation and promote antioxidant protein expression, neuron viability and cerebral blood flow, reduce apoptosis, amyloidogenic effects and loss of dopaminergic neurons [72]. Phenolic acids have been demonstrated to ameliorate epilepsy, neuroinflammation, apoptosis, memory impairments, excitotoxicity and depression, among others [73].
These compounds, apart from their biological activity, stand out for their ubiquity in plants. Their presence in all kinds of fruits, vegetables and plant material has been extensively reported [74,75]. Over the last few years, by-products such as seeds, peels and leaves are gaining attention for their content of bioactive compounds, which is sometimes higher than in the edible parts [76]. However, as mentioned before, these by-products are usually discarded, creating nearly 30% of food waste annually [23].
According to the abovementioned evidence, the administration of phenolic compounds highlights a potential alternative for the treatment of neurodegenerative pathologies. Therefore, the consumption of fruits and vegetables could be therapeutic due to the presence of these phenolic compounds. Their isolation could be a solution, but the direct application of plant extracts could reduce effort, contamination and costs maintaining the same neuroprotective effect, or even more since some extracts exert their effect thanks to the synergistic effect of all their bioactive compounds [77]. Moreover, it would be a great opportunity for the proper revalorization of agri-food by-products, promoting circular bioeconomy of fruit and vegetables.
All pathophysiological mechanisms abovementioned have been studied not only as neurodegenerative disease drivers, but also as feasible therapeutic targets for phenolic compounds and plant extracts rich in phenolics and other bioactive compounds, to exert their neuroprotective activity. In this regard, studies related to this topic are summarized in Table 1. Different in vitro studies where fruit and vegetable by-product extracts rich in phenolic compounds (and other bioactives) were tested in neurodegenerative models. Anti-apoptotic and anti-MMP. [31] A-and B-type procyanidins, rutin, quercetin, saponins.

Plant Species
Suppression of brain inflammation induced by amyloid aggregation.
Modulation of NF-κB signaling pathway. Reduce of mRNA and protein expressions of IL-1, TNF-α, COX-2 AND iNOS. Inhibition of cell apoptosis in microglial cells overactivated by Aβ deposition.
Suppression of brain inflammation induced by amyloid aggregation.
Inhibition of the neuroinflammatory effect induced by amyloid aggregation and deposition by modulation of NF-kB signaling pathway.
Reduced release of proinflammatory cytokines IL-6 and TNF-α from the microglia activation. Growth of neurite length. Induction of mRNA expression of neuronal markers such as MAP2. Decrease in NO production.
Anti-inflammatory. [ AChE and β-secretase inhibition due to hydrogen bond formation.
[88]  Increase in the number of flies capable of flying above the limit established. Anti-AChE. [78] Peel B-type procyanidins, flavanols monomers and chlorogenic acids.
Antioxidant potential in Drosophila melanogaster.
Increase in the life span and locomotor activity, decrease in LPO, prevention of induced locomotor impairment.
Neuroinflammation and protein accumulation in diabetes mice.
Decrease in the inflammatory response and microglia burden, reduction in tau hyperphosphorylation in cortex and hippocampus.
Antioxidant potential and LPO in rats (ex vivo).
Increased expression and activity of SOD and GST. Prevention in LPO upregulation.
Maintenance of mitochondrial homeostasis in Caenorhabditis elegans.
Stimulation of mitophagy through activating protein ULK1, enhancement of the cholinergic neuronal resistance and glutamatergic neurons protection.

Suppression of apoptosis in
Aβ-induced cells through upregulation of Bcl-2 proteins and decrease in caspase-3 mRNA expression.
Anti-apoptotic. [103] Rutin, quercetin, catechin and Proanthocyanidins. BBB dysfunction. Improvement in spatial learning and memory function. Inhibition of caspase-1 and IL-1 expression, and inhibition inflammasome activation through AMPK/mTOR/ULK.  Improvement in episodic memory performance and neural functioning maybe through increased regional perfusion in several brain areas.
Antioxidant potential on albino rats.
Increase in levels of GSH, GPX, SOD and CAT activities. Low MDA levels. Inhibition of LPO. Downregulation of iNOS mRNA expression.
Protein accumulation in a mouse model of AD.
Blockage of Aβ fibril formation interfering with (pre)protofibril formation and oligomerization through β-sheet structure. Reduction in tau aggregations and promotion of α-secretase activity.
Anti-fibrillization, anti-aggregation and anti-secretase. [24,114]   Reduction in amyloid plaque density, increase in neurotrophin BDNF and reduction in AChE activity. Decrease in LPO and in the concentration of pro-inflammatory cytokine TNF-α.
Memory, anxiety and learning in mice. Protein accumulation.
Enhancement of memory, spatial learning and motor coordination. Reduction in anxiety-related behavior through inhibition of the fibrillization of Aβ.
Neuroinflammation and protein accumulation in mice.
Suppression of Aβ levels. Delays in the formation of senile plaques. Reduction in pro-inflammatory ILs and TNF-α expression. Improvement in ATP formation.

Leaf and Flower
Elenolic acid, oleuropein and hibiscus acid.

Apoptosis in Wistar rats (in silico).
Increase in cell viability under oxidative stress conditions. Reduction in mitochondria membrane potential loss, and reversion of caspases to basal levels. Reduced apoptosis mediated by oxidative stress.

Neural precursor cells in mice.
Promotion of cellular survival and neuronal differentiation. Presence of different pro-neurogenic compounds. Increase in neural precursor cell proliferation and neurogenesis. Inducement of endogenous antioxidants.
Pro-neurogenic and antioxidant.
Mitochondrial biogenesis promotion, increase in mitochondrial mass, mtDNA copy number and ATP production. Regulation of AMPK-PGC1α pathway.

Endogenous antioxidant system, LPO and AChE inhibition in Danio rerio.
Enhancement of antioxidant defense by preventing decrease in SOD, CAT, GPX and GSH, and suppressing the increase in MDA and protein peroxidation. AChE inhibition.
Downregulation of enzymes and cytokines in albino rats. Protein deposition.
Diminish inflammatory cascades reducing the neuronal effect of TNF-α. Significant improvement in GSH, SOD, CAT, MDA, NO and AChE levels.
Avoidance of beta-amyloid depositions.

Improvements in flies' mobility.
Decrease in the degeneration of dopaminergic neurons. Anti-PD.

Neuroprotection against Abnormal Protein Deposition
Phenolic compounds have been shown to prevent protein deposition through numerous mechanisms, using one or more at a time. For example, EGCG has been theorized to protect against protein misfolding through antioxidant properties, degrading APP by α-secretase into non-amyloidogenic proteins, direct binding to the protein avoiding its fibrillization, or even preventing tau aggregation and hyperphosphorylation [9,132,133]. Different mechanisms reported for this compound are gathered in Figure 3.

Neuroprotection against Abnormal Protein Deposition
Phenolic compounds have been shown to prevent protein deposition through numerous mechanisms, using one or more at a time. For example, EGCG has been theorized to protect against protein misfolding through antioxidant properties, degrading APP by α-secretase into non-amyloidogenic proteins, direct binding to the protein avoiding its fibrillization, or even preventing tau aggregation and hyperphosphorylation [9,132,133]. Different mechanisms reported for this compound are gathered in Figure 3. Polyphenols are known to reduce β-secretase activity and subsequent production of insoluble proteins, such as oleuropein, quercetin, curcumin and EGCG [9]. Through APP cleavage by α-secretase, these compounds cause the inhibition of the amyloidogenic pathway and protect the CNS from the accumulation of Aβ plaques [9,[132][133][134][135]. Furthermore, phenolic compounds are able to interact with Aβ and αS hydrophobic sequences breaking β-sheet motifs, preventing tau hyperphosphorylation and making proteins smaller and nontoxic [9]. Quercetin, resveratrol, oleuropein, luteolin, myricetin and ECGC, among others, are able to form hydrogen bonds with β-sheet structure, inhibiting the aggregation of Aβ and preventing tau primary conformation self-assembly [132,133,136]. Structural conformation (≥2 phenolic rings and ≥3 OH groups) is considered a more limiting factor to amyloid and synuclein accumulation than oxidative conditions for an optimal 3D conformation for non-covalent bonds with β-sheet structures [101,137].
Other phenolics promote protein solubility modifying their hydrophobicity [138]. Das et al. (2016) reported for the first time that punicalagin (ellagitannin) interacts directly with Aβ motifs to exert neuroprotective effects [136]. Knowing that punicalagin is the major polyphenolic compound in pomegranate extract, the anti-amyloidogenic effects observed by  and Morzelle et al. (2016) of pomegranate peel and pulp extracts could be explained by punicalagin presence [30,123]. The same applies to olive waste such as leaves, whose oleuropein content is enough to potentially be considered interesting extracts with neuroprotective effects, as reported by Chiaino et al. (2020) [41].
Concerning tau hyperphosphorylation, phenolics such as ECGC and myricetin have been shown to block the phenomenon inhibiting the different phases of fibril assembly, e.g., myricetin is able to interfere in the elongation phase [138]. Quercetin can also perform this anti-hyperphosphorylation effect, which is consistent with Infante-García et al.'s Polyphenols are known to reduce β-secretase activity and subsequent production of insoluble proteins, such as oleuropein, quercetin, curcumin and EGCG [9]. Through APP cleavage by α-secretase, these compounds cause the inhibition of the amyloidogenic pathway and protect the CNS from the accumulation of Aβ plaques [9,[132][133][134][135]. Furthermore, phenolic compounds are able to interact with Aβ and αS hydrophobic sequences breaking β-sheet motifs, preventing tau hyperphosphorylation and making proteins smaller and nontoxic [9]. Quercetin, resveratrol, oleuropein, luteolin, myricetin and ECGC, among others, are able to form hydrogen bonds with β-sheet structure, inhibiting the aggregation of Aβ and preventing tau primary conformation self-assembly [132,133,136]. Structural conformation (≥2 phenolic rings and ≥3 OH groups) is considered a more limiting factor to amyloid and synuclein accumulation than oxidative conditions for an optimal 3D conformation for non-covalent bonds with β-sheet structures [101,137].
Other phenolics promote protein solubility modifying their hydrophobicity [138]. Das et al. (2016) reported for the first time that punicalagin (ellagitannin) interacts directly with Aβ motifs to exert neuroprotective effects [136]. Knowing that punicalagin is the major polyphenolic compound in pomegranate extract, the anti-amyloidogenic effects observed by  and Morzelle et al. (2016) of pomegranate peel and pulp extracts could be explained by punicalagin presence [30,123]. The same applies to olive waste such as leaves, whose oleuropein content is enough to potentially be considered interesting extracts with neuroprotective effects, as reported by Chiaino et al. (2020) [41].
Concerning tau hyperphosphorylation, phenolics such as ECGC and myricetin have been shown to block the phenomenon inhibiting the different phases of fibril assembly, e.g., myricetin is able to interfere in the elongation phase [138]. Quercetin can also perform this anti-hyperphosphorylation effect, which is consistent with Infante-García et al.'s (2017) paper. Here, mango leaf extract, reported to possess quercetin and high amounts of polyphenols in its matrix, inhibited tau hyperphosphorylation [102].
Antifibrillization properties from phenolics maintain a close structure-activity relationship [139]. Ortho-dihydroxyl in flat, planar molecules with substituted aromatic end groups covalently binds to αS side chains, remarkably decreasing its hydrophobicity and detoxifying the protein assembly [132,139]. For example, Gu et al. (2017) were able to demonstrate that mulberry extract reduced α-synuclein levels in Lewis bodies in mice with PD [108]. This effect is probably due to the presence of quercetin in the fruit matrix and its capacity to inhibit αS fibrillization [132]. The same applies to Mohammad-Beigi et al.
(2019), who reported olive extract antifibrillization activity and highlighted the extract as a plausible neuroprotective strategy [34].
Finally, Gu et al. (2017) also reported the ability of mulberry extract to upregulate ubiquitin levels, probably due to the high presence of polyphenolic compounds (rutin, quercetin) and different phenolic acids (chlorogenic, caffeoylquinic acids), but with no clear mechanism [108,109]. This probably promotes UPS activity and the normal regulation of proteolysis.
Other studies where the neuroprotective activity from fruit and vegetable by-product extracts are mentioned now. Pasinetti et al. (2010) exposed the role of grape seeds (rich in proanthocyanidins) in blocking Aβ fibril formations by preventing protofibril formation, pre-protofibril oligomerization and initial coiling through β-sheet structure binding, as well as a reduction in tau peptide aggregation [24]. El-Hawary et al. (2021) detailed how different acids from Morus macroura pulp and leaf extract (chrysin, resveratrol and ferulic acid) were able to inhibit abnormal Aβ aggregations, blocking β-secretase activity through conformational features and hydrogen bindings [85]. Additionally, they demonstrated that resveratrol and ferulic acid were able to pass through the BBB and reach their target easily. Main mechanisms are shown in Figure 4.

Neuroprotection against Oxidative Stress
Polyphenols from plants are great antioxidants, mainly due to their capacity to modulate multiple cellular processes, such as redox balance. Knowing how antioxidant systems are affected in neuropathological conditions, the use of plant extract rich in phenolics could ameliorate neurodegenerative processes [140]. The antioxidant potential of phenolic compounds is conducted through three different mechanisms: suppressing enzymes or chelating metal ions which promote ROS, scavenging ROS and upregulating endogenous antioxidative systems [53,55]. Usually, hydroxyl group number and position on the aromatic ring are quite important conditions, which allow flavonoids to be singularly effective antioxidants [55,141]. On the other hand, carotenoids have also been proved to improve the activity of the endogenous antioxidant system, an effect observed by Elseady et al. (2022) in saffron stigma carotenoid-rich extract [53,130].
Polyphenols such as resveratrol, curcumin and ECGC, among others, have been reported to protect against neurodegeneration through the activation of signaling molecular pathways such as Keap1/Nrf-2/ARE, which is the main protective pathway against endogenous and exogenous ROS [140,142]. Here, the Keap1-Nrf-2 complex interacts with the phenolics through the active site of Keap1, provoking its disaggregation and subsequent translocation of Nrf2 to the nucleus, where it triggers the expression of antioxidant molecules such as GSH and HO-1 [140,143]. In addition, another effective mechanism is the inhibition of molecules that are especially sensitive to the oxidative stress state, such as nuclear factor-kB (Nf-kB). Gao et al. (2022) reported an increase in GSH, SOD and CAT activities and a decrease in MDA production after a pre-treatment with Vaccinium dunalianum fruit, leaf and flower extracts, which contained excellent amounts of polyphenols such as kaempferol and feruloylquinic acid [95].
Flavonoids have also been demonstrated to prevent ROS formation by modulating the cell signaling pathway Keap1/Nrf-2/ARE [72]. Apart from that, they are able to reduce amyloidogenic effects, dopaminergic neuron loss, apoptosis, etc. This seems to be related to their ability to form ligands with receptors in the CNS, in addition to the fact that their structure offers them a lipophilic character which allows them to cross the BBB [72]. Nantacharoen et al. (2022) also reported that Cleistocalyx nervosum fruit extract showed antioxidant activity upregulating the gene expression of different cellular antioxidant enzymes, such as SOD, CAT and GPX, promoting the translocation of the Nrf2 protein from the cytoplasm into the nucleus. Additionally, resveratrol found in their extract could be one of the main responsible factors of this effect, as previously reported [86].
Therefore, flavonoid-enriched extracts could show significantly high antioxidant levels, as several studies confirm, either by promoting endogenous antioxidants or scavenging free radicals [75,83,87]. For example, Duangjan et al. (2021) reported that grape leaf extract promoted the gene expression of different cellular antioxidant enzymes such as CAT, SOD, GST and GPX in vivo [87]. Resveratrol, quercetin, apigenin and catechin can be highlighted as the most common reported phenolics in grape leaves, which could be provoking this antioxidant effect. The presence of resveratrol could be the main mechanism through which endogenous antioxidant enzymes levels are promoted, due to the promotion of Nrf2 expression and translocation. Indeed, it was reported that the extract's ability to promote the translocation of DAF-16 in C. elegans, had the same result as Nrf2 translocation.
Other studies demonstrated the ability to downregulate lipid peroxidation, one of the most damaging symptoms of oxidative stress, by reducing levels of MDA [91,123]. All reported biological processes examined are gathered in Figure 4.

Neuroprotection against Neuroinflammation
Polyphenols have been shown to exhibit numerous anti-neuroinflammatory activities. One such example is the downregulation of microglial activation and subsequent release of proinflammatory factors, such as TNF-α, IL-6 and iNOS, through the inhibition of cellular cascade signaling pathways involving NF-kB via nuclear translocation of certain subunits [144,145]. Moreover, there is the inhibition of the activity of COX-2 and LOX-5 enzymes. In third place, there is the attenuation of phosphorylation and overactivation of mitogen-activated protein kinases (MAPKs), such as IkB and p38 MAPK, to protect against dopaminergic neuronal death [144]. Finally, there is the avoidance of TL receptor overexpression, such as TLR4 [145].
The mechanisms through which phenolic compounds prevent chronic neuroinflammation are still scarcely studied. However, the modulation of the NF-kB signaling pathway seems to be one of the most important. NF-kB is a transcription factor that regulates the expression of almost 500 different genes, including pro-inflammatory enzymes and cytokines [146]. Thus, the constant activation of NF-kB in chronic inflammation leads to detrimental effects. Ben Youssef et al. (2021) reported that grape seed and skin extract, especially rich in resveratrol, catechins and gallic acid, inhibited the NF-kB pathway, probably reducing the activity of its subunits [115].
Polyphenols are also theorized to modify morphological features of microglial cells. Tang et al. (2018) found that fully activated microglial cells showed an amoeboid shape. Catechin and procyanidin A2 obtained from lychee seed extracts (LSE) reduced microglia activation by decreasing amoeboid-shaped microglia proportions in cell cultures [80]. These polyphenols were also found to downregulate NF-kB expression and apoptosis. Numerous studies reported this ability to reduce microglia burden and overactivation, reducing the proportion of amoeboid-shaped glial cells [102,120,122]. Zhao et al. (2018) also showed that LSE reduced mRNA levels and the protein expression of pro-inflammatory mediators (IL-1, TNF-α, COX-2, iNOS). Furthermore, the extract was able to attenuate IkB phosphorylation and keep NF-kB inactive [79]. LSE was rich in procyanidins and was probably responsible for the great neuroprotective effect exerted. Phochantachinda et al. (2021) studied the anti-neuroinflammatory potential of Indian gooseberry pulp, which appeared to reduce the release of proinflammatory cytokines IL-6 and TNF-α after inducing microglia activation. Additionally, it promoted neuronal differentiation by inducing the expression of neuronal markers from the MAPK signaling pathway, which modulated the growth and stabilization of microtubules in neurites [48]. Angeloni et al. (2021) reported that coffee by-product extract was able to reduce the activity of pro-inflammatory enzymes, such as iNOS and COX-2, and the levels of different pro-inflammatory mediators by reducing NF-kB proteins to basal levels [89]. In fact, these coffee extracts showed catechin content, in accordance with LSE. A high number of reports have demonstrated brain anti-inflammation through the downregulation of pro-inflammatory cytokines IL-6 and TNF-α, among others, and enzymes such as COX and LOX [42,84,124,129,130]. Key cellular processes are shown in Figure 4. ubiquitin levels, probably due to the high presence of polyphenolic compounds (rutin, quercetin) and different phenolic acids (chlorogenic, caffeoylquinic acids), but with no clear mechanism [108,109]. This probably promotes UPS activity and the normal regulation of proteolysis.

Neuroprotection against Mitochondrial Dysfunctions
Polyphenols are capable of preventing or promoting apoptosis according to necessity, enhancing mitochondrial biogenesis, influencing mitochondrial fission and fusion, affecting mitophagy, controlling mitochondrial quality and regulating mitochondrial functions such as ETC and ATP synthesis [53,54]. The PI3K/Akt/mTOR/AMPK pathway is an interesting biological target for polyphenols to act as both pro-and anti-apoptotic mediators [10,128,147]. Resveratrol has been reported to achieve inhibition of the mTOR complex that competes against ATP for the ATP-binding sites of mTOR [148]. Oleuropein from olives has been also found to trigger autophagy, activating AMPK [149]. Curcumin has been proved to inhibit the Akt/mTOR pathway to induce protective autophagy [147]. Cao et al. (2022) reported that a passion fruit pericarp extract exerted neuroprotective effects through stimulation of mitophagy by inhibiting mTOR and directly activating ULK1 proteins [57]. Passion fruit characterization in previous reports has shown resveratrol and curcumin in its matrix, which could be the reason for this neuroprotective effect [150]. On the other hand, after intoxication or ischemia, neuronal death is harshly triggered throughout the CNS, causing irreparable damage [92]. Therefore, anti-apoptotic activity is needed. Hua et al. (2021) stated that wild turnip root extracts exerted anti-autophagic activity by restoring phosphorylated PI3K, Akt and mTOR levels, which can be translated into an activation of the PI3K/Akt/mTOR pathway and a promotion of cell survival [92].
Wild turnip has been previously reported to possess different phenolic acids in its matrix, such as ferulic and caffeic acids, compounds which have been proven to modulate different signaling cascades related to cell survival and proliferation [151,152]. Bhat et al. (2022) also showed the ability of velvet bean seed extract to promote Akt activity through the PI3K/Akt/mTOR pathway, preventing cell apoptosis and repairing induced damage [97]. Moreover, this extract regulated neurotransmitter levels by reducing MAO activity.
Mitochondria biogenesis (mitogenesis) is a metabolic process dedicated to maintaining the organelle's integrity. Its sensitive relation to oxidative stress turns mitochondria dysfunction into one of the main pathogeneses of neurodegeneration. Polyphenols are able to upregulate the AMPK-PGC1α signaling pathway responsible for mitogenesis: Deng et al. (2019) previously reported on the behavior of the mitochondrial biogenesis promoter shown by ginger extracts, which increased mitochondrial mass, mtDNA copy number and ATP production, as well as the activity of mitochondrial complexes through the regulation of the AMPK-PGC1α signaling pathway [131].
Other studies have focused on different mechanisms. Chiaino et al. (2020) reported increased cell viability by reducing mitochondrial membrane potential loss and cytoplasmatic caspase levels with an extract of olive leaf and hibiscus flower, probably for the presence of oleuropein in the extract [41]. Shin et al. (2021) showed that mulberry fruit extract, rich in phenolic acids and flavonoids, prevented mitochondria membrane from depolarizing, in addition to suppressing pro-apoptotic factors such as cytochrome c [110]. Finally, Elseady et al. (2022) demonstrated the ability of saffron stigma extract to decrease caspase-3 levels, reducing cell apoptosis [130]. All abovementioned is summarized in Figure 4.

Neuroprotection against Impaired Glutamatergic and Cholinergic Neurotransmissions
Excitotoxicity is considered another biological target of polyphenols, which are able to inhibit glutamate-induced Ca 2+ increases and intracellular accumulation by suppressing ion channel proteins through hydrogen-bonding with their amino acid residues, reducing triggering signaling cascades and, finally, reducing glutamate release from their vesicles [153,154]. However, no reports have been found relating this to plant extract effects.
Furthermore, the glutamate accumulation in neurons leads to the generation of intracellular ROS and oxidative stress, as well as the diminution of intracellular GSH levels [86]. Thus, polyphenols may be useful for reducing excitotoxic effects, as Nantacharoen et al. (2022) reported. Here, the Cleistocalyx nervosum extract, rich in resveratrol, was reported to promote the expression of endogenous antioxidant enzymes and suppress the caspase expression in glutamate-induced oxidative damage models [86].
Phenolic compounds also can inhibit enzymatic disorders that end in neurodegeneration, such as the overactivation of AChE and BChE (butyl cholinesterase). In this respect, the inhibition is also explained by morphological features (active sites and oxyanion holes). Polyphenols are able to interact with amino acid residues of enzymes structures through the formation of hydrophobic interactions, π-π and hydrogen bonds thanks to their methoxy and hydroxyl groups [94,155]. This ability is a function of the number and position of their OH groups [94]. Khokar et al. (2021) studied the ability of different phenolic compounds from pomegranate peel extract in silico to attach to AChE and then inhibit its activity. Catechin was reported to be the most promising anti-AD compound for its low binding energy for AChE [88]. Then, other reports of AChE inhibitory activity where the catechin content was high enough could be also explained [67,78,83]. Ben Rejeb et al. (2020) stated that extracts from artichoke by-products (leaves, bracts and stems) inhibited AChE and BChE activity, and they related this to the presence of flavonoids in a positive and significant way [93]. In fact, they highlighted luteolin and its derivatives as the main contributing factor, and reported that flavonoids such as quercetin were able to block the entrance to the active site of AChE and BChE enzymes.
Some studies were conducted ex vivo, recollecting brain tissue after the consumption of the studied extract, such as Phachonpai et al. (2021), who used wampee fruit peel to attenuate memory deficits in rats and showed that cognitive impairment was reduced by its supply [106]. Main mechanisms and biological processes are gathered in Figure 4.

Conclusions and Future Perspectives
AD and PD are the most common neurodegenerative conditions suffered by humans as they grow older, and represent major unmet challenges for medicine. In the present scenario, plant by-products have been deeply highlighted as natural antioxidant extracts that allegedly are able to display neuroprotective effects thanks to their phenolic compoundrich compositions. Although this effect has been extensively exposed and reported in different samples, the use of plant extracts as neuroprotective agents is still limited. Main limitations are now exposed: • Digestion, absorption and metabolization of fruit, vegetables and other plants are crucial for recovering phenolic and other bioactive compounds from their matrices. Their bioavailability is considered one of the most limiting factors for plants to exert neuroprotection, since their effects are positively related to the amount consumed [1,156]. Low absorption rates and quick metabolism could limit their efficacy, but abusive consumption could lead polyphenols to show pro-oxidant activity.

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Circulating metabolite structures usually differ from their native dietary compound, so it could be thought that their activity also differs [18]. Additionally, levels of circulating bioavailable metabolites are always below consumed levels, limiting their neuroprotection even more [157].

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Secondary metabolites from phenolic compounds should be able to pass the BBB to reach the brain. Therefore, their effectiveness also depends on their permeability capacity across this barrier [12]. • Phenolic compounds are not the only compounds in plant matrices, so neuroprotection could be exerted by others too, and may be under synergistic or antagonistic interactions. • Difficulties in carrying out in vivo studies in humans.
These questions, among others, remain unresolved. Therefore, more research should be conducted in order to elucidate the mechanisms and physiological processes through which fruits, vegetables and plants exert their neuroprotection. Several solutions are currently in progress, such as the encapsulation of plant extracts using biodegradable materials, which promotes the enhancement of phenolic bioavailability and BBB permeability, and their therapeutic efficacy [132].
After this review, results obtained establish a starting point for further research that must be carried out in order to confirm plant extracts' beneficial effects. Here, a detailed description of the neuroprotective nature of different phenolic compounds and extracts has been exposed, demonstrating their value for the development of innovative neuroprotective strategies.