Tannic Acid Attenuates Quinolinic Acid-Induced Neurodysfunction by Modulating Oxidative Stress Parameters and Electrogenic Pump Activity

doi:10.21203/rs.3.rs-4810458/v1

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Tannic Acid Attenuates Quinolinic Acid-Induced Neurodysfunction by Modulating Oxidative Stress Parameters and Electrogenic Pump Activity

https://doi.org/10.21203/rs.3.rs-4810458/v1

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The pathophysiology of neurodegenerative illnesses is largely dependent on oxidative stress and poor ion homeostasis, and these conditions represent a substantial worldwide health burden. Endogenous neurotoxic quinolinic acid (QA) is linked to neurodysfunction by inducing oxidative stress and interfering with sodium pump function. In a number of models, the polyphenolic molecule tannic acid (TA), which has strong antioxidant qualities, has demonstrated pharmacological effects in several diseased conditions. However, the neuroprotective effect of TA is rather speculative and still very open for clarification. In the present study, an in vitro model was employed to examine the effect of TA on deoxyribose degradation, lipid peroxidation, thiol status, antioxidant enzymes and cerebral and spinal sodium pump in rat cerebral and spinal tissue homogenates treated with quinolinic acid (QA, 2 mM). Results revealed that QA treatment led to a profound (p < 0.05) degradation of deoxyribose, formation of thiobarbituric acid reactive substances (TBARS), and marked (p < 0.05) reduction in tissue level of free thiols. However, TA treatment significantly (p < 0.05), counteracted TBARS production, deoxyribose degradation and markedly (p < 0.05) increased the thiol level of the cerebral and spinal tissue homogenates. Furthermore, QA markedly (p < 0.05) diminished the activities of cerebral and spinal antioxidant enzymes [catalase (CAT), superoxide dismutase (SOD), Glutathione Peroxidase (GPx) and Glutathione S transferase (GST)] and impaired the activities of cerebral and spinal sodium pump. Nonetheless, the activities of the antioxidant enzymes and pump were all raised in both the cerebral and spinal tissue homogenates upon TA treatment. These findings justify the pharmacological action of TA on QA-induced neurotoxicity and suggest its potential use in the treatment of neurodegenerative diseases. Further investigation is required to determine TA's translational usefulness in clinical settings.

Tannic acid

quinolinic acid

neurodysfunction

antioxidants enzymes

thiols

lipid peroxidation

Na+/K+-ATPase

A disease known as neurodysfunction occurs when an agent that is chemical, biological, or physical can negatively impact the structure or functionality of the central and peripheral nervous systems (Cunha-Oliveira et al., 2008). It happens when a person is exposed to a chemical, specifically a neurotoxin or neurotoxicant, which modifies the nervous system's typical activity and damages neural tissue permanently or irreversibly (Cunha-Oliveira, 2008).Over time, this can damage or even kill neurons, which are the cells that send and process impulses in the brain and other parts of the nervous system. Organ transplants and pro-oxidant exposures, including radiation, specific medication regimens, recreational drug use, heavy metal exposure, bites from specific venomous snake species, pesticides (Keifer et al., 2007; Costa et al., 2008), specific industrial cleaning solvents (Sainio and Markku, 2015), fuels (Ritchie et al., 2001), and certain naturally occurring substances, can all result in neurotoxicity.

The onset of symptoms may come on quickly or later after exposure. These could include delusions, headaches, uncontrollably obsessive or compulsive behaviors, limb weakness or numbness, loss of memory, eyesight, and intellect, as well as cognitive and behavioral issues and sexual dysfunction. Neurological illnesses and neurodegenerative diseases are becoming increasingly a great menace to world health. For example, the World Health Organization (WHO) reported that over 55 million people worldwide suffered from dementia in 2019, and it is predicted that this figure will increase to approximately 139 million by 2050. Neural illnesses have been linked to oxidative stress, which is defined by an imbalance between antioxidant defense mechanisms and the formation of reactive oxygen species (ROS).

Neurodegeneration, the progressive loss of neuronal structure or function brought on by neurotoxic chemicals, is the root cause of neurodegenerative disorders. Cell damage or death may result from such neuronal damage in the end. Amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, multiple system atrophy, tauopathies, and prion disorders are examples of neurodegenerative diseases. Neurodegeneration arises at several levels of neuronal circuitry in the brain, from molecular to systemic. These conditions are regarded as incurable since there is no known method to stop the neurons' ongoing deterioration. Numerous subcellular commonalities between these illnesses have been identified by biomedical research, such as increased cell death and abnormal protein assemblages (such as proteinopathy) (Rubinsztein, 2006; Bredesen et al., 2006). These parallels imply that improvements in treatment for one neurodegenerative illness may also benefit others.

Quinolinic acid (QUIN or QA) is a dicarboxylic acid having a pyridine backbone that is also referred to as pyridine 2,3-dicarboxylic acid. It is a solid with no color. It is the kynurenine pathway's metabolic precursor of niacin (Hiroshi et al., 2011). According to Guillemin et al. (2003), quinolinic acid exhibits a strong neurotoxic impact. Research has indicated that quinolinic acid may play a role in a variety of mental illnesses, as well as neurodegenerative brain processes and other ailments. Quinolinic acid is only generated by activated macrophages and microglia in the brain. Quinolinic acid has been classified as an endogenous excitotoxin since it is derived from the central nervous system and has the potential to seriously harm neurons (Schwarcz et al., 1983). According to recent studies, quinolinic acid can cause lipid peroxidation in the mammalian brain and spine, which may result in the production of free radicals (Rios and Santamaria, 1991).

When prooxidants cause lipid peroxidation, the membrane lipids are frequently subjected to oxidative damage from free radical attack. A number of disorders, particularly neurodegenerative conditions like Parkinson's disease, include lipid peroxidation as a key pathogenic mechanism. Antioxidants can, however, prevent or delay cellular damage despite these oxidative stress-related etiologies primarily because of their ability to scavenge free radicals. These low-molecular-weight antioxidants have the ability to safely interact with free radicals, stopping the chain reaction before it causes harm to important components. Glutathione, ubiquinol, and uric acid are a few examples of antioxidants that the body naturally produces during regular metabolism (Shi et al., 1999). The diet contains other, milder antioxidants. Additionally, research in the fields of biochemistry and pharmacology has demonstrated the significant functions that polyphenols, such as tannic acid, play in the absorption and neutralization of free radicals, the quenching of singlet and triplet oxygen, and the breakdown of peroxides produced by pro-oxidants like QA (Jing et al., 2002).

Taking into account the fact that neurological disorders and neurodegeneration are significantly influenced by oxidative stress (Stephenson, 2018; Singh et al., 2019; Pereira, 2021), tannic acid's antioxidant ability as a naturally occurring polyphenolic molecule thus provides a fascinating avenue for further research into its possible neuroprotective properties and the body of evidence pointing to a connection between oxidative stress and quinolinic acid-induced neurotoxicity. However, little is known about how specifically tannic acid reduces this neurotoxicity.This research was therefore undertaken to evaluate the attenuating potential of tannic acid against quinolinic acid induced neurodysfuction in rat cerebral and spinal tissues.

2.1 Chemicals

The following materials were purchased from Sigma (St. Louis, MO): tannic acid (TA), thiobarbituric acid (TBA), 2-deoxyribose, quinolinic acid, sodium hydroxide, 5,5-dithiobis-2-nitrobenzoic acid (DTNB), trichloroacetic acid (TCA), acetate buffer, Tris-HCl buffer, sodium dodecyl sulphate (SDS), adenosine triphosphate (ATP), ouabain, and quinolinic acid (QA). The remaining chemicals were procured from regular commercial suppliers and were of analytical quality.

2.2 Experimental animals

Wistar rats weighing between 200-250g were used for the experiment at Biochemistry departmental animal house. During this period, they were maintained at room temperature and provided with standard feed, water and light cycle in accordance with Faculty of Pure and Applied Sciences ethical guide for care and use of laboratory animal, Federal University Wukari, Nigeria.

2.3 Tissue Preparation

The animals were given a light ether anesthesia, decapitated, and their organs (the spine and brain) were removed before being promptly frozen and weighed. The organs were properly cleaned to remove any blood stain by rinsing them in cold 50 mM Tris-HCl buffer. The tissue was homogenized right away using a homogenizer in a cold, pH 7.4 (1/10, w/v) 50 mM Tris-HCl solution. The low-speed supernatant fraction, which was utilized for the tests, was obtained by centrifuging the homogenate at 4000 x g for ten minutes.

2.4 Deoxyribose Degradation

The amount of deoxyribose degradation was measured in accordance with published data (Halliwell et al., 1987). Hydroxyl radicals break down deoxyribose and generate reactive compounds known as thiobarbituric acid (TBA). For 30 minutes, deoxyribose (2 mM) was incubated at 37 ⁰C with 50 mM potassium phosphate (pH 7.4), 2 mM QA to cause deoxyribose breakdown, and TA (concentration of 0–10 µM). Following the incubation period, the tubes were heated for 20 minutes at 100 ⁰C upon addition of 0.4 ml of TBA 0.8% and 0.8 ml of TCA 2.8% and spectrophotometric measurements were made at 532 nm.

2.5 Thiobarbituric Acid Reactive Species (TBARS) Assay

By measuring TBARS (Thiobarbituric acid reactive species) using a modified Ohkawa et al. (1979) technique, lipid peroxidation was measured. An aliquot of 100 µl of S1 was incubated in a reaction system with 50 mM Tris-HCl buffer (pH 7.4), 100 µl of tissue homogenate, and [QA (2 mM)] for one hour at 37⁰C with TA (final concentrations range of 0–10 µM). 200 µl of 8.1% SDS (Sodium Dodecyl Sulphate) was added to the reaction mixture containing S1 to initiate the color reaction. This was followed by the addition of 500 µl of pH 3.4 acetate buffer and 500 µl of 0.8% TBA. For thirty minutes, this combination was incubated at 100 ⁰C. TBARS production was detected under UV-visible light at 532 nm.

2.6 GSH level

Following Ellman's (1959) method of deproteinization with TCA (5% in 1 mmol EDTA), the levels of GSH in the brain and spinal cord were measured using Ellman's reagent. The evaluation of the pancreas' antioxidant capability also made use of the deproteinized brain and spinal tissues.

2.7 Antioxidant enzymes activities

100 µl of tissue homogenate was incubated for 1hour at 37⁰C in the presence of TA (final concentrations range of 0–10 µM), in a reaction system containing 50 mMTris-HCl buffer (pH7.4), with [QA (2 mM)]. The resulting aliquots were then used for the assay of Catalase (CAT), Glutathione S-transferase (GST), Superoxide dismutase (SOD) and Glutathione peroxidase (GPx) activities.

Catalase

Using the Aebi (1984) approach, which entails tracking the disappearance of H₂O₂ in the presence of the homogenate at 240 nm, the catalase activity was measured spectrophotometrically. In a solution containing 50 mM phosphate buffer, pH 7.0, an aliquot and the substrate (H₂O₂) were added to a final concentration of 0.3 mM to start the enzymatic reaction. The units used to express the enzymatic activity were 1 U, which breaks down 1 µmol of H₂O₂ per minute at pH 7 at 25 ⁰C.

Glutathione S -transferase activity

The activity of glutathione S-transferase (GST) was measured in accordance with Habig et al. (1974). In short, for a standard experiment, a suitable quantity of enzyme was added to a reaction mixture of 3 mL that included a final concentration of 100 mM potassium phosphate buffer (pH 6.5), 1 mM GSH, and 1 mM CDNB.

Superoxide dismutase (SOD) activity

The nitrobluetetrazolium (NBT) method developed by Beauchamp and Fridovich (1971) was used to measure SOD activity. O₂ reduces NBT to blue formazan, which absorbs strongly at 560 nm. This process is inhibited by the presence of SOD. The assay mixture was composed of 50 µl of aliquot, 1.5 mg/ml BSA, 0.75 mM NBT, 3 mM EDTA, and 3 mM xanthine in 0.05 M sodium carbonate buffer (pH 10.2). After adding 50 µl of xanthine oxidase (0.1 mg/ml) and letting it sit at room temperature for 30 minutes, the reaction was started. After adding 6 mM CuCl₂ to halt the reaction, the mixture was centrifuged for 10 minutes at 1,500 rpm and the absorbance of blue formazan in the supernatants was measured at 560 nm.

Glutathione peroxidase (GPx) activity

GPx was measured using the Paglia and Valentine (1967) method. 0.1 M phosphate buffer (pH 7.0), 1 mM EDTA, 10 mM glutathione (GSH), 1 mM NaN₃, 1 unit of glutathione reductase, 1.5 mM NADPH, and 0.1 ml of homogenate were all included in the reaction mixture. Following a 10-minute incubation period at 37⁰C, 1 mM of H₂O₂ was added to each sample. At 340 nm, the rate of NADPH oxidation was used to calculate GPx activity.

2.8 Sodium Pump Assays

An aliquot of S1 was utilized in the Na⁺/K⁺ -ATPase activity tests. In a final volume of 500 µl, the reaction mixture included pro-oxidants [QA (2 mM)] and TA (final concentrations range of 0–10 µM), 3 mM MgCl, 125 mM NaCl, 20 mM KCl, and 50 mM Tris-HCl, all at pH 7.4. A final dose of 3.0 mM of ATP was added to start the process. The same conditions were used for the controls, but ouabain (0.1 mM) was added. The Fiske and Subbarow (1925) method was used to measure the amount of released inorganic phosphate (Pi). The difference between two assays was used to compute the Na⁺/K⁺ -ATPase activity (with and without ouabain). Every experiment was carried out a minimum of three times, yielding comparable outcomes and the activity of enzyme was expressed as number of moles of phosphate (P_i) released min^− 1mg protein^− 1.

2.9 Statistical Analysis

The collected data were all presented as mean ± SEM. When appropriate, Duncan's multiple range tests were conducted after the data were analyzed using an appropriate ANOVA. At p < 0.05, means having distinct superscripts differ considerably.

3.1 Effects of tannic on quinolinic acid induced deoxyribose degradation

As displayed below in Fig. 1, quinolinic acid caused a profound (p < 0.05) degradation of deoxyribose sugar. However, the tannic acid exerted marked inhibitory effect (p˂0.05) on deoxyribose degradation in a concentration dependent manner.

3.2 Effects of tannic acid on quinolinic acid induced lipid peroxidation in rat brain and spine homogenates

The results in Fig. 2 revealed that quinolinic acid induced a marked increase in lipid peroxidation in the cerebral and spinal tissue homogenates. However, the tannic acid elicited a marked (p˂0.05) inhibitory effect on lipid peroxidation in a concentration dependent manner compared to the control.

3.3 Effects of tannic acid on GSH level in quinolinic acid-treated rat brain and spine homogenates

The results in Fig. 3 revealed that quinolinic acid caused a profound decrease of GSH level in rat brain and spine homogenates. However, treatment with tannic acid raised the GSH level (p˂0.05) in a concentration dependent manner compared to the control.

3.4 Effects of tannic acid on Na ⁺ /K ⁺ -ATPase activity in quinolinic acid-treated rat brain and spine homogenates

As shown in Fig. 4 quinolinic acid inhibited the activity of the cerebral and spinal Na⁺/K⁺-ATPase, but treatment with tannic acid restored Na⁺/K⁺-ATPase activity (p˂0.05) in a concentration dependent manner compared to the control.

3.5 Effects of tannic acid on antioxidant enzymes activities in quinolinic acid-treated cerebral and spinal tissue homogenates of rats.

Tables 1 and 2 shows the effects of tannic acid on antioxidants enzymes in QA treated cerebral and spinal tissue homogenates respectively. QA caused a marked (p < 0.05) depletion in the level of SOD, CAT, GPx and GST. In the brain tissue homogenate (Table 1). However, treatment with TA elicited a profound (p < 0.05) increase in the activities of these enzymes. Similarly, as revealed in Table 2, the activities of SOD, CAT, GPx, GST in the spinal tissue homogenate were profoundly (p < 0.05) declined by QA, but the administration of TA exhibited substantial (p < 0.05) increase in their activities.

Table 1

Effects of tannic acid on antioxidant enzymes activities in quinolinic acid-treated cerebral tissue homogenates of rats.
Treatment Tubes	CAT	SOD	GPx	GST
A Control	647.11 ± 88.08^a	31.64 ± 8.37^a	31.21 ± 3.85^a	3.13 ± 0.09^a
B QA	422.74 ± 85.26^b	13.57 ± 2.53^b	14.02 ± 2.17^b	0.97 ± 0.03^b
C QA + 1 µM TA	438.53 ± 77.38^b	19.48 ± 2.99^c	17.84 ± 3.07^b	1.21 ± 0.06^c
D QA + 2 µM TA	426.83 ± 69.29^b	19.86 ± 4.5^c	15.55 ± 2.53^b	1.19 ± 0.08^c
E QA + 4 µM TA	522.28 ± 95.83^c	17.93 ± 4.41^c	21.91 ± 2.99^c	3.53 ± 0.51^d
F QA + 6 µM TA	539.3I ± 83.05^cd	23.84 ± 3.83^d	26.51 ± 3.51^d	2.92 ± 0.22^a
G QA + 10 µM TA	579.64 ± 65.28^d	27.59 ± 4.85^d	24.16 ± 2.57^cd	4.26 ± 0.35^e
Data are shown as the mean ± standard deviation (SD) of three separate tests. At p < 0.05, means with distinct superscripts exhibit a significant difference.
SOD = Superoxide dismutase (nmol/min/mg protein)
CAT = Catalase (µmole/mg protein/min)
GST = Glutathione S-transferase (µmole/mg protein/min)
GPx = Glutathione peroxidase (µmole/mg protein/min)

Table 2

Effects of tannic acid on antioxidant enzymes activities in quinolinic acid-treated spinal tissue homogenates of rats.
Treatment Tubes	CAT	SOD	GPx	GST
A Control	228.35 ± 31.93^a	7.84 ± 0.75^a	7.68 ± 0.59^a	0.55 ± 0.03^a
B QA	121.53 ± 18.35^b	2.99 ± 0.53^b	2.07 ± 0.06^b	0.173 ± 0.07^b
C QA + 1 µM TA	119.73 ± 13.53^b	3.65 ± 0.46^c	1.38 ± 0.10^b	0.39 ± 0.02^c
D QA + 2 µM TA	144.63 ± 15.08^c	6.27 ± 0.46^d	6.22 ± 0.12^c	0.35 ± 0.04^c
E QA + 4 µM TA	139.22 ± 12.83^c	4.14 ± 0.51^c	4.74 ± 0.58^d	0.33 ± 0.04^c
F QA + 6 µM TA	128.63 ± 9.52^c	6.48 ± 0.29^d	6.85 ± 0.41^c	0.39 ± 0.01^ac
G QA + 10 µM TA	194.11 ± 24.37^a	6.26 ± 0.39^d	6.65 ± 0.33^c	0.37 ± 0.02^c
Data are shown as the mean ± standard deviation (SD) of three separate tests. At p < 0.05, means with distinct superscripts exhibit a significant difference.
SOD = Superoxide dismutase (nmol/min/mg protein)
CAT = Catalase (µmole/mg protein/min)
GST = Glutathione S-transferase (µmole/mg protein/min)
GPx = Glutathione peroxidase (µmole/mg protein/min)

Tannic acids, one of the many naturally occurring antioxidants, has been shown to have a variety of biological effects, such as antiviral, antibacterial, anti-inflammatory, antiallergic, antithrombotic, and vasodilatory properties. Antioxidant activity is actually a basic characteristic essential to life, and it can lead to the development of antiaging, anticarcinogenic, and antimutagenic properties, among others (Cook and Samman, 1996). Tannic acid and other phenolic compounds have antioxidant action primarily because of their redox characteristics, which enable them to function as hydrogen donors, reducing agents, and singlet oxygen quenchers. Moreso, they might have the ability to chelate metals (Liyana-Pathirana and Shahidi, 2006; Rice-Evans, 1995).

One of the components of DNA, deoxyribose, is released as cells die and their DNA breaks down. (Asamari and colleagues, 1996; Aruoma and colleagues, 1990; Gutteridge, 1984). Interaction with ROS can cause a wide range of oxidative modifications to DNA, such as damages to the deoxyribose moiety of the DNA double helix's sugar-phosphate backbone, nucleobase modifications within the sequence, single- and double-strand breaks and DNA-protein crosslinks (Toyokuni et al., 1995). During normal metabolic circumstances, these kinds of oxidative damage to DNA are typically repaired by the cells; but, during oxidative stress, the volume of DNA lesions exceeds the capacity of the repair process, leaving some DNA lesions unrepaired.

These unrepaired DNA lesions have the potential to cause oxidative stress-related etiologies, such as neurodegenerative disorders. One metabolite of the kynurenine pathway that has been linked to the pathophysiology of neurodegenerative illnesses is quinolinic acid, which may be a neurotoxin. Concerns regarding its potential to contribute to neuronal injury are raised by its capacity to cause oxidative stress in neural tissues (Singh et al., 2019; Pereira, 2021). Figure 4.1 illustrates a significant (p˂0.05) increase in degradation of deoxyribose after quinolinic acid treatment of deoxyribose sugar.This therefore, depicts that quinolinic acid could cause the degradation of deoxyribose constituents of nucleic acid present in the brain and spinal tissue homogenates. However, treatment with tannic acid significantly (p˂0.05) inhibited the degradation of deoxyribose by QA. Hence, tannic acid could exhibit potent inhibitory actions against degradation of deoxyribose components of the nucleic acid in the rat brain and spinal tissue homogenates and this result is in tandem with the one demonstrated by Kade et al. (2007), that tannic acid inhibits degradation of the deoxyribose by ferrous sulphate (Feso₄).

One of the main mechanisms of CNS injury caused by free radicals is lipid peroxidation, which directly destroys neuronal membranes and produces a variety of downstream products that cause significant cellular damage. Highly reactive electrophilic aldehydes, such as malondialdehyde (MDA), 4-hydroxy-2-nonenal (HNE), the most prevalent product, and acrolein, the most reactive, are formed when free radicals attack polyunsaturated fatty acids (PUFAs) (Esterbauer et al., 1991; Pryor and Porter, 1990; Loidl-Stahlhofen et al., 1994). As displayed in Fig. 4.2, quinolinic acid evoked a marked (p˂0.05) production of lipid peroxidation adducts in both brain and spinal tissue homogenates. Increased membrane stiffness, decreased activity of membrane-bound enzymes (such as the sodium pump), damage to membrane receptors, and changed permeability are all consequences of peroxidation of membrane lipids (Anzai et al., 1999; Yehuda et al., 2002). Apart from causing harm to phospholipids, radicals have the ability to target membrane proteins directly and create crosslinks between proteins and lipids, which are linked to modifications in the integrity of the membrane. It is plausible to speculate that disruption of all the aforementioned functions exhibited by PUFAs and their metabolites, in conjunction with protein modification, impact neuronal homeostasis and thus contribute to brain and neuronal dysfunction.

However, treatment with tannic acid exerted an inhibitory effect on the cerebral and spinal lipid peroxidation.

Researchers have shown that the primary components with antioxidant and antiradical qualities are flavonoids and flavones, which include polyphenols like tannic acids (Poterat, 2017). Tannic acid may therefore be useful in the treatment of chronic diseases whose origin is connected to oxidative stress. This agrees with the findings of Azimullah et al. (2023) that tannic acid mitigated ROS and MDA induced by rotenone. In many organisms including animals, glutathione (GSH) functions as an antioxidant. Important cellular constituents can be shielded from harm by glutathione from sources such heavy metals, peroxides, free radicals, and reactive oxygen species (Pompella et al., 2003). As shown in Fig. 4.3, the delivery of QA to rat brain and spinal tissue homogenates resulted in a significant (p˂0.05) reduction in GSH levels. However, treatment with TA markedly (p˂0.05) increased GSH levels. This suggests that by raising their level in the central nervous system, tannic acid may modulate the cellular antioxidant system in counteracting the oxidative attack elicited by QA. This also aligns with the work of Azimullah et al. (2023) that tannic acid abolished the depletion of GSH induced by rotenone.

Moreover, animal cells' plasma membranes are maintained by the sodium pump, an enzyme that is membrane bound. The ability to adjust to shifting physiological and cellular stimuli depends on this enzyme (Therien and Blostein, 2000). This protein is highly produced by neurons and is responsible for maintaining the electrical potential required for the excitability of this tissue by using between 30 and 60% of the brain's ATP reserve. Quinolinic acid profoundly inhibited the action of Na⁺/ k⁺-ATPase in the homogenate of the brain and spine, as Fig. 4.4 illustrates. Conversely, treatment with tannic acid restored the activity of the enzyme at high concentrations. This result correlates with the one demonstrated by Kade et al. (2013), that tannic acid could profoundly (p˂0,05) restore the activity of Na⁺/ k⁺-ATPase in the cerebral tissue homogenate following reduction of its activity by streptozotocin. Thus this suggest that tannic acid could have some pharmacological effects on the central nervous system.

A collection of antioxidant enzymes that can prevent oxidative damage brought on by pro-oxidants make up the antioxidant pool. SOD, CAT, GST, and GPx are some of these enzymes. First line of defense against injury mediated by reactive oxygen species (ROS) is formed by SOD (Kangralkar et al., 2010). These proteins lower the level of superoxide anion free radical (O₂), which damages cells at high concentrations, by catalyzing its dismutation into molecular oxygen and hydrogen peroxide (H₂O₂). The GPx are a type of antioxidant enzymes that are related to heme-free thiol peroxidases, just like peroxidases. They help to reduce the toxicity of H₂O₂ and organic hydroperoxides by catalyzing their reduction to water or matching alcohols (Zhao et al., 2019).

Similarly, one of the most significant antioxidant enzymes is catalase. Almost all aerobic organisms include it. In a two-step procedure, catalase converts two molecules of hydrogen peroxide into one molecule of oxygen (Ossowski et al., 1993) and two molecules of water (Deisseroth and Dounce, 1970). A family of multifunctional enzymes known as the GSTs is responsible for the well-established detoxification of electrophilic metabolites and xenobiotics (Mannervik and Danielson, 1989; Hayes and Pulford, 1995; Zimniak and Singh, 2006). The conjugation of a broad range of structurally different molecules with electrophilic carbon, nitrogen, or sulfur atoms to GSH is generally catalyzed by GSTs. This study further investigated how tannic acid could restore enzymatic antioxidants in the brain and spinal tissue homogenates after quinolinic acid had reduced their activity. Table 4.1 shows that QA administration profoundly (p˂0.05) impaired the activity of SOD in the cerebral tissue homogenate. Nevertheless, tannic acid treatment resulted in a significant (p < 0.05) rise in the activity of the enzyme. Fridovich et al. (1989) indicate that this response implies tannin may mitigate total oxidative stress by successfully restoring cellular defenses against superoxide radicals. Furthermore, quinolinic acid administration caused a significant (p < 0.05) drop in CAT activity in the brain tissue homogenate.

On the other hand, catalase activity is significantly (p ~ 0.05) increased upon tannic acid treatment. According to (Kade et al., 2013), this points to a possible mechanism by which tannic acid supports hydrogen peroxide detoxification by protecting cells from oxidative damage. Furthermore, in cerebral tissue homogenate, tannic acid demonstrated a considerable (p < 0.05) ability to restore GPx activity following its depletion caused by quinolinic acid. This suggests that tannic acid can help the body's defense mechanism against free radicals by encouraging the breakdown of lipid and other organic peroxides. Moreover, quinolinic acid significantly (p < 0.05) reduced the cerebral tissue homogenate's GST level. Tannic acid administration, however, demonstrated a profound (p < 0.05) rise in GST activity. In similar manner, as displayed in table 4.2, QA markedly (p˂0.05) diminished the activities of CAT, SOD, GST and GPx in the spinal tissue homogenate and this effect is significant (p˂0.05) when compared to the control. Nonetheless, tannic acid substantially (p˂0.05) abolished the detrimental effect of QA by evoking a marked (p˂0.05) increase in the activities of the spinal antioxidant enzymes.

This reveals that tannic acid increases oxidative stress in order to support the antioxidant pool within cells by augmenting the efficacy of the antioxidant enzymes. Findings herein are similar to those of Azimullah et al. (2023) that demonstrated that tannic acid mitigated the depletion antioxidants, SOD, CAT, and GSH, accompanied with profound MDA and NO. Apparently, it is worth mentioning that tannic acid has the efficacy to counteract oxidative damage and ameliorate or restore sodium pump activity in neurological conditions.

The results here in, revealed that tannic counteracted the detrimental effects of QA in rat cerebral and spinal tissue homogenates by restoring sodium potassium pump activity and modulating antioxidant indices. Consequently, it is rational to conclude that TA could mitigate oxidative stress processes, redox and homoeostatic complication that characterize neurological diseases. Tannic acid could be considered for the treatment and management of neurodegenerative diseases due to its observed antioxidant and pharmacological activity. However, further investigation is required to determine TA's translational efficacy in clinical settings.

ACKNOWLEDGEMENT

We would like to express our sincere gratitude to all authors for their invaluable contributions to this manuscript. Their expertise, dedication and collaborative spirit greatly enriched the quality of this work. The research is borne out of authors’ financial contribution as there was no funding from any agency.

Ethical Approval

This study was carried out according to standard guideline of the Committee on Care and Use of Experimental Animal Resources of Faculty of Pure and Applied Sciences, Federal University Wukari, Nigeria with the approval number, FUW/FPAS/23/021.

Consent to Participate

Not applicable

Consent to Publish

Not applicable

Competing Interest

The authors have no relevant financial or non-financial interests to disclose

Authorship contribution statement

Ale, E.M conceptualized and designed the research. Ayo, V.I. supervised the work. Asuelimen, S.O. carried out data analysis and editing. Material preparation and data collection were performed by Timothy, M.J. Visualization and resources procurement was carried out by Adewole, M.A. Nathan, R.N. carried out project administration, while Ale, E.M. wrote the first draft of the manuscript. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

Data Availability

Authors will provide all data for the research on request

Aebi, H. (1974). Catalase. In: Bergmeyer, H.U., Ed., Methods of Enzymatic Analysis, Verlag Chemie/Academic Press Inc., Weinheim/NewYork, 673-680.
Anzai, K., Ogawa, K., Goto, Y., Senzaki, Y., Ozawa, T., Yamamoto, H. (1999). Oxidation-dependent changes in the stability and permeability of lipid bilayers. Antioxid. Redox Signal. 1:339–347; 1999.
Aruoma, O.I., Chaudhary, S.S., Grootreld, M., Halliwell, B. (1990). Binding of iron (II) ions to pentose sugar 2-deoxyribose. J. Inorg. Chem. 35, 149–155.
Asamari, A. M., Addis, P. B., Epley, R. J., Krick, T.P. (1996). Wild rice hull antioxidants. J. Agric. Food Chem. 44, 126–130.
Azimullah, S., Meeran, M.F.N., Ayoob, K., Arunachalam, S., Ojha, S., Beiram, R. (2023). Tannic Acid Mitigates Rotenone-Induced Dopaminergic Neurodegeneration by Inhibiting Inflammation, Oxidative Stress, Apoptosis, and Glutamate Toxicity in Rats. Int. J. Mol. Sci. 24: 9876. https://doi.org/10.3390/ ijms24129876
Beauchamp, C., Fridovich, I. (1971). Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44:276.
Bredesen, D. E., Rao, R. V., Mehlen, P. (2006). "Cell death in the nervous system". Nature. 443 (7113): 796–802.
Cook, N.C., Samman, S., 1996. Flavonoids: chemistry, metabolism,cardioprotective effects and dietary sources. J. Nutr. Biochem. 7, 66
Costa, Lucio, G., Giordano, G., Guizzetti, M., Vitalone, A. (2008). "Neurotoxicity of pesticides: a brief review". Front. Biosci. 13 (13), 1240–9.
Cunha-Oliveira, T., Rego, A.C., Oliveira. C.R. (2008). "Cellular and molecular mechanisms involved in the neurotoxicity of opioid and psychostimulant drugs". Brain Res. Rev. 58 (1), 192–208
Deisseroth A., Dounce A. L. (1970). Catalase: physical and chemical properties, mechanism of catalysis, and physiological role. Physiol. Rev.;50(3):319-375. doi: 10.1152/physrev.1970.50.3.319
Esterbauer, H., Schaur, R. J., Zollner, H. (1991). Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic. Biol. Med. 11:81–128.
Fiske, C. H., Subbarow, Y. J. (1925) The colorimetric determination of phosphorus. J. Biol. Chem. 66:375–381.
Fridovich, I. (1983). Superoxide radical: an endogenous toxicant. Annu Rev Pharmacol Toxicol 23: 239–257.
Guillemin G. J., Croitoru-Lamoury J., Dormont D., Armati P. J., Brew B. J. (2003a). Quinolinic acid upregulates chemokine production and chemokine receptor expression in astrocytes. Glia 41 371–381. 10.1002/glia.10175
Gutteridge, J. M. (1984). Reactivity of hydroxyl and hydroxyl-like radical discriminated by release of thiobarbituric acid-reactive material from deoxyribose, nucleosides and benzoate. Biochem. J., 224, 761–767
Habig, W. H., Pabst, M. J., Jakob, W. B. (1974). Glutathione S-transferases the first enzymatic step in mercaptyric acid formation J. Biol. Chem.; 249: 7130 – 39.
Halliwell, B., Gutteridge, J.M.C., Aruoma, O.I. (1987). The deoxyribose method: a simple ‘‘Testtube’’ assay for determination of rate constants for reactions of hydroxyl radicals. Anal Biochem, 165:215–9.
Hayes, J.D., Pulford, D.J. (1995). The glutathione S-transferase super gene family: regulation of GST and contribution of the isozymes to cancer chemoprevention and drug resistance. Crit Rev Biochem Mol Biol. 30:445–600.
Hirose, Y., Watanabe, K., Minami, A., Nakamura, T., Oguri, H., Oikawa, H. (2011). Involvement of common intermediate 3-hydroxy-L-kynurenine in chromophore biosynthesis of quinomycin family antibiotics. J. Antibiot. (Tokyo). 64, 117–122. doi: 10.1038/ja.2010.142
Jing, W., Xiaolan, C., Yu, C., Feng, Q. (2022). Pharmacological effects and mechanisms of tannic acid. Biomed. Pharmacother. 154(Suppl 1):11356. DOI:10.1016/j.biopha.2022.113561
Kade, I. J., Johnson, D. O., Akpambang, V.O.E., Rocha, J.B.T. (2012). Polymerization of gallic acid enhances its antioxidant capacity: Implications for plant defence mechanisms. Biokemistri, 24(1):15-22
Kade, I.J., Balogun, B.D., Rocha, J.B.T. (2013). In vitro glutathione peroxidase mimicry of ebselen is linked to its oxidation of critical thiols on key cerebral sulphydryl proteins – A novel component of its GPx-mimic antioxidant mechanism emerging from its thiol-modulated toxicology and pharmacology. Chem Biol Interact; 206(1):27–36.
Kangralkar, V.A., Patil, S.D., Bandivadekar, R.M. (2010). Oxidative stress and diabetes: A review. Int. J. Pharmacol. Appl.; 1:38-45.
Keifer, Matthew C., Firestone, J. (2007). "Neurotoxicity of Pesticides". J. Agromed.. 12 (1), 17–25
Liyana-Pathirana, C.M., Shahidi, F., 2006. Antioxidant properties ofcommercial soft and hard winter wheats (Triticum aestivum L.) andtheir milling fractions. J. Sci. Food Agric., 86:477.
Loidl-Stahlhofen, A., Hannemann, K. and Spiteller, G. (1994). Generation of alphahydroxyaldehydic compounds in the course of lipid peroxidation. Biochim. Biophys. Acta, 1213:140–148.
Mannervik, B., Danielson, U.H. (1989). Glutathione S-transferases-structure and catalytic activity. CRC Crit Rev Biochem Mol Biol. 23:283–337.
Ohkawa, H., Ohishi, H., Yagi, K, (1979). Assay for lipid peroxide in animal tissues by thiobarbituric acid reaction. Anal Biochem. 95:351–8. Ellman, G. L. (1959). Tissue sulfhydryl groups. J. Biochem. Biophys., 82:70 – 7.
Ossowski, V.I., Hausner, G. and Loewen, P. C. (1993). “Molecular evolutionary analysis based on the amino acid sequence of catalase,” J. Mol. Evol. 37; 71–76.
Paglia, D. E., Valentine, W. N. (1967). Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med.; 70:158-69.
Pereira, T. M. C., Côco, L. Z., Ton, A. M. M., Meyrelles, S. S., Campos-Toimil, M., Campagnaro, B. P., Vasquez, E. C. (2021). "The Emerging Scenario of the Gut-Brain Axis: The Therapeutic Actions of the New Actor Kefir against Neurodegenerative Diseases". Antioxidants. 10 (11), 1845.
Pompella, A., Visvikis, A., Paolicchi, A., De Tata, V., Casini, A.F. (2003). The changing faces of glutathione, a cellular protagonist. Biochem. Pharmacol., 66(8): 1499-1503
Pryor, W. A., Porter, N. A. (1990). Suggested mechanisms for the production of 4- hydroxy-2-nonenal from the autoxidation of polyunsaturated fatty acids. Free Radic. Biol. Med 8:541–543.
Rice-Evans, C., 1995. Plant polyphenols: free radical scavengers orchain breaking antioxidants? Biochem. Soc. Symp. 61:103.
Rios C., Santamaria A. (1991). Quinolinic acid is a potent lipid peroxidant in rat brain homogenates. Neurochem. Res.16:1139–1143.
Ritchie, G.D., Still, K.R., Alexander, W.K.N., Alan F. (2001). "A review of the neurotoxicity risk of selected hydrocarbon fuels". J. Toxicol. Environ. Health B Crit. Rev. 4 (3), 223–312.
Rubinsztein, D. C. (2006) "The roles of intracellular protein-degradation pathways in neurodegeneration". Nature. 443 (7113): 780–6.
Sainio, Markku. and Alarik. (2015). "Neurotoxicity of solvents". Occupational Neurology. Handb. Clin. Neurol.; (131), 93–110.
Schwarcz, R., Whetsell, W.O. Jr, Mangano, R.M. (1983). Quinolinic Acid: An Endogenous Metabolite That Produces Axon-Sparing Lesions in Rat Brain. Sci., 219(4582):316-318 DOI: 10.1126/science.6849138
Shi HL, Noguchi N, Niki N. Comparative study on dynamics of antioxidative action of α- tocopheryl hydroquinone, ubiquinol and α- tocopherol, against lipid peroxidation. Free Radic Biol Med. 1999;27:334–46.
Singh, A., Kukreti, R., Saso, L., Kukreti, S. (2019). "Oxidative Stress: A Key Modulator in Neurodegenerative Diseases". Mol. 24 (8): 1583.
Stephenson, J., Nutma, E., van der Valk, P., Amor, S. (2018). "Inflammation in CNS neurodegenerative diseases". Immunol. 154 (2): 204–219
Therien, A.G., Blostein, R. (2000). Mechanisms of sodium pump regulation Am J Physiol Cell Physiol, 279(3):C541-66. doi: 10.1152/ajpcell.2000.279.3.C541.
Toyokuni, S. O., K.; Yodoi, J.; Hiai, H. (1995). FEBS Lett. 358, 1-3.
Yehuda, S., Rabinovitz, S., Carasso, R. L., Mostofsky, D. I. (2002). The role of polyunsaturated fatty acids in restoring the aging neuronal membrane. Neurobiol. Aging 23:843–853.
Zhao, L., Zong, W., Zhang, H., and Liu, R. (2019). Kidney toxicity and response of selenium containing protein-glutathione peroxidase (Gpx3) to CdTe QDs on different levels. Toxicol. Sci. 168 (1): 201–208.
Zimniak, P., Singh, S.P. (2006). Families of glutathione transferases. In: Awasthi YC, editor. Toxicology of glutathione transferases. CRC Press; Boca Raton, FL:11–26

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Tannic Acid Attenuates Quinolinic Acid-Induced Neurodysfunction by Modulating Oxidative Stress Parameters and Electrogenic Pump Activity

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Abstract

Figures

1 INTRODUCTION

2 MATERIALS AND METHOD

2.1 Chemicals

2.2 Experimental animals

2.3 Tissue Preparation

2.5 Thiobarbituric Acid Reactive Species (TBARS) Assay

2.6 GSH level

2.7 Antioxidant enzymes activities

Catalase

Superoxide dismutase (SOD) activity

Glutathione peroxidase (GPx) activity

2.8 Sodium Pump Assays

2.9 Statistical Analysis

3 RESULT

3.1 Effects of tannic on quinolinic acid induced deoxyribose degradation

4 DISCUSSION

5 CONCLUSION

Declarations

References

Supplementary Files

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