Trazodone attenuates the neuroinflammation signaling mechanism and restores the activity of BACE 1 and ChAT in scopolamine-induced AD-like symptoms in male wistar rats

doi:10.21203/rs.3.rs-3766910/v1

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Trazodone attenuates the neuroinflammation signaling mechanism and restores the activity of BACE 1 and ChAT in scopolamine-induced AD-like symptoms in male wistar rats

https://doi.org/10.21203/rs.3.rs-3766910/v1

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Trazodone, an antidepressant may play a role in attenuating neuroinflammation induced by scopolamine. We propose the potential novel mechanistic insights that link up between neuroinflammation and memory biomarkers modulate the pathogenesis of AD. Docking studies and histopathological alterations were performed against scopolamine toxicity. Scopolamine was administered into the brain through the peritoneal cavity for the establishment of the AD model. Trazodone (TRAZ) was administered orally used to treat AD-like symptoms in male wistar rats aged between 200–250 g. We confirmed that TRAZ administration significantly attenuates the SCOP-induced cognitive decline targeted neuroinflammation-mediated age-dependent disease progression. TRAZ attenuates the activity of TNF-α, ChAT, and BACE1 and inhibits apoptosis. These findings confirmed that TRAZ has the potential efficacy to modulate the interference in cognitive decline induced by scopolamine. TRAZ is the controlling candidate that regulates the neuroinflammation-dependent memory biomarkers activity response pathway that positively manages apoptosis.

Trazodone

Neuroinflammation

Memory Biomarkers

Apoptosis

Neuroprotection

The central nervous system (CNS) is the primary target of neuroinflammation in the pathophysiological event of Alzheimer’s disease (AD). Currently, few promising targets have been reported to reduce neuroinflammation and the mechanistic interplay between microglia and neuronal cells in treating AD. The inflammation is the secondary response followed by any injury or infection which confirms that neuroinflammation is the key component in neurodegenerative disorders like AD [1]. Due to the complex and multifactorial etiology of AD pathogenesis [2–4], neuroinflammation plays a fundamental role in etiopathogenesis [5, 6], due to the ability to worsen Aβ and tau protein [7]. Age-dependent pathogenesis is specialized by deteriorating immune system [8] and imbalance of homeostasis [9, 10] which is highly involved in damaging mitochondria [11, 12], impaired autophagy, and mitophagy [13, 14] along with dysregulated ubiquitin-proteasome system (UPS) unit [15, 16] and activates DNA-damaging response [17, 18]. Modifications in immune biomarkers found in the blood include CRP, TNF-α, IL-6 [19–21], and its soluble receptors of TNF-α [22, 23]. All these receptors and biomarkers together have a crucial role in the development of disease [24, 25, 26]. Glucose metabolism, oxidative stress, and synaptic loss are also triggered by mitochondrial dysfunctioning and contribute to AD pathogenesis [27, 28, 29, 30, 31]

Age progression is the strongest irreversible phenomenon to cause AD [32, 33]. The lack of effective therapies for AD treatment leads to high damage to memory and cognitive activities [34]. AD is the second leading cause of dementia, which will affect 6.7 million people of Americans over the age of 65 in 2023 [35]. The well-known reported mechanistic insights to cause the pathogenesis of AD, are the accumulation of Aβ protein extracellularly and tau protein hyperphosphorylation intracellularly [36–38]. The reported hypothesis of amyloid stat + ed that proteolytic cleavages of amyloid precursor protein by BACE 1 enzyme is the main mechanism of Aβ production and is an effective target for disease modification [39]. The AD-like model established by inducing scopolamine (SCOP) is frequently used to examine cognitive disease. Memory and cognitive function depend heavily on cholinergic activity. According to the cholinergic theory, cognitive impairment in AD patients is mostly caused by the destruction of cholinergic neurons in the brain. Now, it is well established that cholinergic neuronal circuit dysfunction significantly contributes to the cognitive deficits linked to aging and neurodegenerative illnesses like AD [40]. SCOP traditionally blocked working memory-related muscarinic acetylcholine (ACh) receptors [41]. To address memory loss and cognitive impairment in mild and moderate AD, cholinergic agonists, and acetylcholinesterase (AChE) inhibitors have been developed [42]. The process of programmed cell death, or apoptosis, is crucial for tissue homeostasis and health growth [43]. However inappropriate apoptosis has been linked to neurodegenerative conditions like AD [44, 45]. Learning and memory are severely impacted by apoptosis of neuronal tissue [46, 47]. DNA fragmentation is a key component of apoptosis and the assay TUNEL uses terminal deoxynucleotidyl transferase to identify it [48]. Trazodone (TRAZ) was docked into the AChE enzyme and its expected binding mode was predicted to understand how it functions as an AChE inhibitor and interacts with the target. The discovery that TRAZ may pass through the active site of the enzyme and attach to the AchE site demonstrates the medication’s potential as a therapeutic lead or molecular probe for the treatment of AD. An effective antidepressant with a long history of usage, TRAZ is especially helpful for treating insomnia and agitation in AD patients. In the 1960s era of TRAZ (FDA-approved) for the treatment of depression, was widely used off-label to treat anxiety, schizophrenia, etc [49–52]. The behavioral and psychological signs and symptoms of AD are lessened by the TRAZ treatment [50, 51]. Thus, we aimed to establish the effects of TRAZ on cognition and to determine whether there was evidence that it could be used in the treatment of neurodegenerative diseases and whether reduces neuroinflammation or not [52].

2.1. In-silico-Docking

The program package AutoDock vina (version 1.2.3, the most recent version available) was used for molecular docking. First, the ligand’s 3D conformers were retrieved in. sdf format from PubChem, and energy was reduced using ChemBio3D Ultra 14.0. The RCSB PDB website allowed users to download the crystal structures of proteins or receptors. The proteins were created by using Swiss PDB Viewer to add the missing residues and atoms, MolProbity to add hydrogens, and AutoDock Tools (version 1.5.7., the most recent) to add polar hydrogens and Kollman charges. The generation of the configuration file is considered the ligand binding site. Grid points with a 0.375 Å spacing for each protein’s binding sites were used to create the configuration file. The AutoDock Vina, which took a minimum of 9 conformers into account, was selected for the best conformer searches. The ligand interactions module of BIOVIA/Discovery Studio Visualizer 2021 software was used to visualize the binding interactions between the chemicals and the protein.

2.2. Drugs and Chemicals

All the required chemicals and reagents used in present research were of analytical grade and were obtained from various manufacturers. Scopolamine (#1610001) and trazodone hydrochloride (#PHR3372) were bought from Sigma-Aldrich (St. Louis, MO, USA).

2.3. Ethical Approval and Experimental Design

Male wistar rats weighing between 220–250 g were purchased from CAHF, Jamia Hamdard, New Delhi, India, by Arrive guidelines. The experimental setup received complete IAEC approval (Registration No. 173/G0/Re/D/2000/CPCSEA). Eight rats from each experimental group [control, scop (3 mg/kg; i.p.) only, scop (3 mg/kg; i.p.) + TRAZ (10 and 30 mg/kg, p.o.) were used to evaluate the anti-inflammatory effect of TRAZ against scop-induced AD-like model. At CAHF, all animals are kept in comfortable cages made of polycarbonate and filled with a dry husk. The rats were kept in enclosures that were kept at ambient temperature (22 ± 3ᵒc, 55–60%). The doses were freshly prepared and dissolved in nuclease-free water. The dose regimen and route of administration were based on published literature with slight modifications. Figure 1 provides a schematic illustration of the experimental design.

2.4. Histological studies

2.4.1. Hematoxylin and eosin staining (H & E)

The cornerstone of anatomical pathology diagnosis is the tissue section stained with hematoxylin and eosin (H&E) To early distinguish cellular components, the H&E technique uses contrasting colors to stain the nucleus and the cytoplasm. Rat brains were perfused and then fixed (n = 8) with 4% paraformaldehyde. By using H&E staining, the pyknotic nuclei in the rat prefrontal cortex (PFC) were identified and examined. For staining, the PFC slices of the brain were cut 3 µm thick and mounted on glass slides (Salman et al., 2022). Images were taken with a Zeiss microscope (ZEN Blue edition) at a magnification of 20. Using ImageJ software, the number of pyknotic neurons was counted.

2.4.2. Cresyl violet (CV) staining

Neurons in the brain and spinal cord can be stained with cresyl violet (CV). It is employed to show the presence of the nissl substance in the cell nuclei and neurons. Rat brains were stained with CV to enable the visual identification of nissl bodies in the PFC area of the brain. To facilitate subsequent processing, the sections were cut into 3µm thick and kept on glass slides. After 10 minutes of staining with CV solution, the paraffin-embedded section was dehydrated gradually using graded alcohol. A Zeiss microscope (ZEN Blue Edition) was used to take pictures with a 20x magnification [53].

2.5. Immunohistochemistry studies

Immunohistochemistry was performed with the formalin-fixed paraffin-embedded slides of the brain. 3µm thick paraffin sections proceeded to deparaffinization and hydration steps and were subsequently exposed antigen retrieval in citrate buffer by heating (pH 6.0). They were perfused with 4% paraformaldehyde and sacrificed. The brain tissues were carefully washed with 0.1 M phosphate buffer isolated the PFC region and treated with 3% H₂O₂ to block peroxidase activity. The PFC sections were blocked with 2% goat serum in 0.1 M PBS for 1 h at room temperature (RT) [53]. After that, the PFC sections were incubated at 4ᵒC overnight with rabbit polyclonal antibody against TNF-α (1:100, #D2D4; CST) and BACE 1 (1:100, #D10E5; CST), ChAT (1:100, #GTX113164) in continue with the incubation with a secondary antibody. After the development with DAB chromogen, the sections were observed under 20×magnifcation; Zeiss microscope (ZEN Blue edition).

2.6. Immunofluorescence study

An essential immunochemical technique known as immunofluorescence (IF) enables the identification and localization of numerous antigens in diverse cell preparations and tissues. The brain's formalin-fixed, paraffin-embedded slides were used for immunofluorescence. Deparaffinization and hydration processes were performed on 3µM thick paraffin sections before they were exposed to antigen retrieval in citrate buffer (pH 6.0). Slides should be incubated with Triton-X (0.3%) for 30 minutes at room temperature before being rinsed three times with buffer. Following a wash, slices were incubated for an hour at room temperature in a blocking solution containing 1.5% (wt/vol) bovine serum albumin (BSA) [54]. Phosphate buffer was used to dilute the primary antibodies (Aβ, CST, and D3D2N), which were then incubated at 4 ^oc overnight. By three times rinsing the sections in 1 x phosphate buffer, the unbound antibody was eliminated. After being diluted 1:200 in phosphate buffer, the secondary antibody (DyLight 488, goat anti-mouse IgG) was applied to the sections and let to sit for an hour at room temperature. Slides were mounted with 4′,6-diamidino-2-phenylindole (DAPI, stock solution 10 mg/ml, diluted 1:10,000) for counterstaining of nuclei (Fluoroshield; GTX30920) and examined under a microscope after being washed with buffer and mounted.

2.8. Statistical Analysis

The present study signified in comparisons between scop and treated groups of TRAZ were performed using one-way analysis of variance (ANOVA), in continue with Tukey's multiple comparison tests. Statistical study was supported using GraphPad Prism 8.0 (GraphPad Software, San Diego, CA, USA). Values were expressed as mean ± s.e.m and the nominal level of significance was identified at p < 0.05.

3.1. In-silico characterization of TRAZ against AchE

The docking score of TRAZ with AChE protein (PDB ID: 4M0E) was found to be -10.448 kcal/mol. The 2D and 3D interaction of TRAZ with AChE protein is depicted in Fig. 3 and Fig. 4, respectively. It fits into the binding pocket of the AChE protein containing amino acids Trp86, Gly121, Gly122, Tyr124, Glu202, Ser203, Trp286, Val294, Phe295, Phe297, Tyr337, Phe338, Tyr341, and His447. It showed various types of interactions like conventional hydrogen-bonding, Pi-alkyl, Pi-Pi T-shaped, and van der Waals interactions. The keto group of the ligand forms hydrogen bonding with the amino acid His447 whereas the amino acids Trp86, Trp286, and Tyr341 are involved in Pi-alkyl bonding with the ligand. The aromatic and triazole rings of the ligand interact with the amino acids Trp286 and Tyr337, respectively via Pi-Pi T-shaped bonding

3.2. TRAZ attenuates BACE1 activity for AD Treatment

BACE 1 or β-secretase and γ-secretase are the key players producing Aβ by the cleavage of amyloid precursor protein. Researchers focusing on the BACE 1 inhibitor to reduce the production of Aβ [55]. Our results have shown that the activity of BACE 1 is significantly increased in the scop-group (***p˂0.001) as compared to the control. The TRAZ treatment significantly reduces the expression of BACE 1 (^###p˂0.001) as compared to scop-induced AD-like symptoms and indicates that TRAZ has the potential to attenuate the BACE 1 activity [Figure 3B]. Data was displayed as mean ± SD, followed by ANOVA with Tukey’s multiple comparison test, where n = 5.

3.3.TRAZ reduced the Aβ₄₂ expression

Aβ₄₂ (the most toxic species) and the main culprit reported till now to cause AD. It has shown a variety of harmful insights by modifying and altering the multiple pathways. Aβ₄₂ can cause excitotoxicity, mitochondrial damage, neuroinflammation, etc [56]. Our results have shown a pronounced increase in Aβ₄₂-positive expression in the scop-induced group (^**p˂0.01) as compared to control (dark blue) and anti-Aβ₄₂ antibody (green). Both, the TRAZ-treated groups showed a significant decrease in the Aβ₄₂ expression as compared to the scop-induced group (^##p˂0.01) [Figure 4B]. Data was displayed as mean ± SD, followed by ANOVA with Tukey’s multiple comparison test, where n = 5.

3.4. Cholinesterase deficiency associated with AD

In rat models of AD, memory loss is a result of cholinergic circuit degradation in the PFC area. Using a scop-induced AD-like model, we examined how the cholinergic circuit is activated and how it affects learning and memory [57]. The arrow indicates the expression (Fig. 3A). The ChAT expression was significantly lower in the scop-induced AD-like model as compared control (**p˂0.01). The dose-treated group of trazodone (10 and 30 mg/kg) significantly increased the expression of ChAT (^##p˂0.01; ^###p˂0.001) (Fig. 5B). Data was displayed as mean ± SD, followed by ANOVA with Tukey’s multiple comparison test, where n = 5.

3.5. High expression of TNF-α is correlated with AD pathogenesis

In the rat PFC region, TNF-α (cytokine) expression was measured and its relation to AD prognosis was investigated [58]. We found that TRAZ administration attenuated the expression of cytokine and reduced inflammation. The arrow indicated the expression of TNF-α in the PFC region of the brain (Fig. 2A). The higher the expression, the higher will be the inflammation. The scop-induced experimental group has significantly increased expression as compared to control (*p˂0.05). Moreover, the TRAZ-treated groups (10 and 30 mg/kg) significantly reduced the expression of cytokine (^###p˂0.001; ^###p˂0.001), TNF-α in comparison to the scop-induced AD-like model (Fig. 6B). Data are presented as mean ± SD, and statistical examination by applying one-way ANOVA repeated measures with Tukey’s multiple comparison test, where n = 7, at 20X magnification.

3.6. Restoration of damaged neurons by the treatment of TRAZ

To identify the histological alterations, we performed the hematoxylin & and eosin staining in the PFC region and quantified the pyknotic neurons after scop-induction. TRAZ administration attenuates the histopathological alterations after scop-induction. The arrows indicate the pyknotic neurons in the regions of the PFC (Fig. 4A). The number of pyknotic neurons in the scop-induced group was (***p˂0.001) as compared to the control. TRAZ (10 and 30 mg/kg) restored the damaged neurons significantly (^###p˂0.001) in comparison to the scop-induced AD-like group (Fig. 7B). Here, data are displayed as mean ± s.e.m and one-way ANOVA repeated measures was applied with Tukey’s multiple comparison tests where n = 9, at magnification 20X.

3.7. TRAZ treatment attenuated neuronal damage

The normal neurons were intact with a clear border and a light stain. ATOR administration reduces neuronal damage after scop-induction. The arrows indicate the morphology of neurons in the PFC region in rat brain tissue (Fig. 5A). The number of damaged neurons in PFC regions was, (***p˂0.001) as compared to the control. TRAZ (10 and 30 mg/kg) restored the damaged neurons significantly (^###p˂0.001) in comparison to the scop-induced AD-like group (Fig. 8B). Here, data are signified as mean ± s.e.m and one-way ANOVA repeated measures alongwith Tukey’s multiple comparison tests where n = 7, at magnification 20X.

This study is the first to identify the effectiveness of TRAZ against AChE by in-silico characterization. AutoDock Vina software is used to evaluate ligand interaction against the targeted protein. AChE (target protein) is a highly demanding interest in the current market of AD drugs [59]. Our in-silico study revealed the highest binding score of TRAZ against AChE and indicates that TRAZ has the potential to treat cognition. We further investigate the efficacious role of TRAZ in a scopolamine-induced AD-like rodent model. The already-known fact is that scopolamine is the chemical inducer model to creates AD-like symptoms affects cognition and produces neuroinflammation [60]. Our present study highly indicated that TRAZ is the best candidate to attenuate cytokine; TNF-α. The immunohistochemistry parameters showed significantly increased expression of TNF-α in the scopolamine group as compared to control and attenuated by the treatment of TRAZ. We confirmed from the present study that TRAZ has the capability to combat neuroinflammation against scopolamine by using immunohistochemistry.

BACE 1 enzyme concentrations and their sparked activity in AD pathogenesis support the hypothesis that BACE 1 is the key player in producing Aβ. Several researchers are focusing on finding BACE 1 inhibitors for slowing disease progression. The highly advanced and demanding approach of BACE 1 inhibitors makes it a therapeutic approach for combating AD [60]. We studied the qualitatively expressed BACE 1 and quantitatively estimated the count of its expression in the PFC region of rat brains. And, confirmed that TRAZ significantly reduces the rate of BACE 1 as compared to scopolamine in the PFC region of the brain. Our result indicates that TRAZ has the potential to make a BACE 1 inhibitor and combat AD-like symptoms. Moreover, this study, to the best of our knowledge, determined the modulatory activity of ChAT protein against the treated group. Immunohistochemistry revealed qualitatively and quantitively that, the expression and count of ChAT protein are significantly reduced in the treated group of rats, specifically in the region of PFC. ChAT activity in the AD-like group was established by inducing scopolamine has been significantly attenuated by the potential efficacy of TRAZ. Both, BACE 1 and ChAT are modulated to combat AD-like symptoms and prove the effectiveness of TRAZ in cognition.

The well-known inducer of AD-like symptoms “Scopolamine” is also causing damage to the neurons revealing histopathologically. Our present findings showed that TRAZ is the best candidate to recover or reduce the number of pyknotic nuclei and damaging neurons in the PFC region of rat brain confirmed by hematoxylin & eosin and cresyl violet stainings.TRAZ treatment still falls short of the standards for successful treatments following AD etiology. The fact that the current investigation exclusively examined the preventative impact of TRAZ in male Wistar rats poses substantial limitations because both sexes can be involved in the etiology of AD and females are more common. At certain biological points, males, and females both display the same amount of variability [38]. However, only male rats were shown to have cognitive impairments and other clinical symptoms of AD [39]. Additionally, TRAZ is a candidate for anti-depressants. AD affects cognitive and non-cognitive symptoms which include memory, locomotor activity, etc. In this study, we did not perform any locomotor test or not to check the muscle strength. The development of the novel strategy might be supported by a clearer understanding of the etiology of AD illness symptoms. When researching neuroprotective mechanisms, sex differences in the incidence of AD should also be considered as they may point to the necessity for distinct therapy modalities. Therefore, additional research is required to understand how behavioral and molecular pathways in both males and females with AD models contribute to another neuroprotective aim of TRAZ. We conclude that a markedly increased depression may raise the commencement of the etiology of AD. The reported study has developed that TRAZ is a potential candidate for AD-like symptomatic treatment. TRAZ proved to be an excellent symptoms-modifying therapeutic intervention by promoting the inhibition of neuroinflammation, the activity of BACE 1 and ChAT, and promoting the recovery at tissue level.

AChE-Acetyl choline esterase

ACh-Acetyl choline

Aβ- Amyloid Beta

AD- Alzheimer disease

ANOVA- Analysis of Variance

BACE 1- β-Site amyloid precursor protein cleaving enzyme 1

BSA- Bovine serum albumin

CAHF- Central animal house facility

CHAT- Choline acetyltransferase

CPCSEA- Committee for the Purpose of Control and Supervision of Experiments on Animals

CRP- C-reactive protein

CV- Cresyl violet

DNA- Deoxyribonucleic acid

FDA- Dood and Drug administration

H & E- Hematoxylin and Eosin

IAEC- Institutional Animal Ethics Committee

IF- Immuno- fluoresce

IL-6- Interleukin

mAChRs- Muscarinic cholinergic receptors

PFC- Prefrontal cortex

SCOP- Scopolamine

TNF-α- Tumor necrosis factor alpha

TRAZ- Trazodone hydrochloride

TUNEL- Terminal deoxynucleotidyl transferase dUTP nick end labeling

Acknowledgement

The authors also would like to thank DST FIST (SR/FST/LS-I/2017/05[C]) and DST PURSE grant (SR/PURSE Phase 2/39[C]) for their support. Prof. Suhel Parvez appreciates the Central Council for Research in Unani Medicine (File. No.3-51/2022-CCRUM/Tech) for their support. Ms. Neha received the Senior Research Fellowship from DST through the INSPIRE program (File No. DST/INSPIRE/03/ 2019/001769. IVR No. 201900031405). The authors would like to thank the funding agencies, ICMR Senior Research Fellowship (File No. 45/14/2020/PHA/BMS) awarded to Ms. Pinky.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Data Availability Statement

All data generated or analyzed during this study are included in this published article.

Ethics Statement

The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC) of Jamia Hamdard (Registration No. 173/ GO/ReBi/S/2000/CPCSEA).

Conflict of Interest Statement

There is no conflict of interest for any of the authors.

Author Contribution

Conceptualization: N, P & S.P.; Methodology: N, P.; P.D., Software: N, P; Validation: N., P., & S.P.; Formal analysis: N, P, P.D., & S.P.; Investigation: N, P & S.P.; Resources: S.P.; Data curation: N, P & S.P.; Writing-original draft: N, P & S.P.; Writing-review & editing: N, P & S.P.; Visualization: N, P & S.P.; Supervision: S.P.; Project administration: S.P.; Funding acquisition: S.P.

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No competing interests reported.

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Graphical Abstract: The muscarinic cholinergic receptors (mAChRs) are antagonistic to the anticholinergic medication scopolamine, which can lead to impairments in the processes of learning acquisition and consolidation. Alzheimer's disease (AD) is a chronic neurological condition for which there is no known cure. Because it participates in the rate-limiting process that results in the toxic oligomeric Aβ₄₂ peptides that deposits in the form of plaques extracellularly and commenced AD, β-secretase (BACE1) is seen as a prospective and sensible target. Scopolamine is used to produce symptoms like AD or to slow cognitive deterioration. Lack of cholinergic activity linked to anti-inflammatory, anti-oxidant, and anti-apoptotic actions. Increased inflammation, including elevated levels of pro-inflammatory cytokines like TNF-α, remains a key factor in explaining the pathogenesis of neurocognitive disorders even though dysregulation of the cholinergic system and oxidative stress have been promising hypotheses for explaining the pathogenesis of AD.

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Trazodone attenuates the neuroinflammation signaling mechanism and restores the activity of BACE 1 and ChAT in scopolamine-induced AD-like symptoms in male wistar rats

Status:

Version 1

Abstract

Figures

1. Introduction

2. Material And Methods

2.1. In-silico-Docking

2.2. Drugs and Chemicals

2.4. Histological studies

2.4.1. Hematoxylin and eosin staining (H & E)

2.4.2. Cresyl violet (CV) staining

2.5. Immunohistochemistry studies

2.6. Immunofluorescence study

2.8. Statistical Analysis

3. Results

3.1. In-silico characterization of TRAZ against AchE

3.2. TRAZ attenuates BACE1 activity for AD Treatment

3.3.TRAZ reduced the Aβ₄₂ expression

3.4. Cholinesterase deficiency associated with AD

3.5. High expression of TNF-α is correlated with AD pathogenesis

3.6. Restoration of damaged neurons by the treatment of TRAZ

3.7. TRAZ treatment attenuated neuronal damage

4. Discussion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

Status:

Version 1

Trazodone attenuates the neuroinflammation signaling mechanism and restores the activity of BACE 1 and ChAT in scopolamine-induced AD-like symptoms in male wistar rats

Status:

Version 1

Abstract

Figures

1. Introduction

2. Material And Methods

2.1. In-silico-Docking

2.2. Drugs and Chemicals

2.4. Histological studies

2.4.1. Hematoxylin and eosin staining (H & E)

2.4.2. Cresyl violet (CV) staining

2.5. Immunohistochemistry studies

2.6. Immunofluorescence study

2.8. Statistical Analysis

3. Results

3.1. In-silico characterization of TRAZ against AchE

3.2. TRAZ attenuates BACE1 activity for AD Treatment

3.3.TRAZ reduced the Aβ42 expression

3.4. Cholinesterase deficiency associated with AD

3.5. High expression of TNF-α is correlated with AD pathogenesis

3.6. Restoration of damaged neurons by the treatment of TRAZ

3.7. TRAZ treatment attenuated neuronal damage

4. Discussion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

Status:

Version 1

3.3.TRAZ reduced the Aβ₄₂ expression