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PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This association between inflammatory signaling and neurotrophic modification establishes a crucial mechanistic link between chronic inflammation and synaptic dysfunction in AD (Thakur et al. 2023; Colavitta and Barrantes 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Oxidative stress further contributes to BDNF suppression and neuronal dysfunction in LPS-induced neuroinflammation (Correia et al. 2024; Shen et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our preliminary comparative analysis revealed that 3D spheroid cultures exhibit more pronounced inflammatory responses compared to traditional 2D cultures, a finding that merits deeper mechanistic discussion.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Several interconnected factors contribute to this enhanced inflammatory reactivity in three-dimensional neuronal models.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The enhanced inflammatory response observed in 3D spheroids is predominantly attributable to their capacity to replicate the complex cell–matrix interactions characteristic of the brain microenvironment.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
In contrast to two-dimensional (2D) cultures, where cells attach to rigid plastic substrates, three-dimensional (3D) spheroids enable cells to interact with self-produced extracellular matrix (ECM) components, including laminin, collagen IV, and proteoglycans.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These components closely approximate the native brain ECM composition (Pereira et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This three-dimensional configuration has been shown to promote mechanotransduction signaling, whereby cells respond to mechanical signals through integrin-mediated pathways, which may enhance inflammatory responses (Boncristiani et al. 2025).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The brain's distinctive ECM composition, characterized by its low elastic modulus (~ 1 kPa) and high levels of hyaluronic acid and chondroitin sulfate proteoglycans, generates a microenvironment that modulates glial activation and neuroinflammatory cascades (Rueda-Gensini et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
In 3D spheroids, the soft, hydrogel-like matrix environment facilitates more authentic cell-ECM interactions, which can amplify TLR4 signaling and downstream inflammatory pathways, as compared to the artificial rigidity of 2D substrates (Valiuliene et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The three-dimensional geometry of spheroids has been demonstrated to increase intimate cell–cell interactions and to permit improved paracrine signaling networks that intensify inflammatory responses.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
In the context of three-dimensional models, the close physical proximity of neuronal cells has been demonstrated to promote more efficient intercellular communication through gap junctions and direct cell-surface receptor contacts (Bae et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This enhanced connectivity facilitates the expeditious dissemination of inflammatory signals, encompassing cytokine cascades and Danger-Associated Molecular Patterns (DAMPs), throughout the spheroid structure.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Recent research reveals that 3D neurosphere models exhibit higher expression of connexin proteins and improved gap junction functionality compared to 2D cultures, facilitating coordinated cellular responses to inflammatory stimuli (Stefano et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This synchronized inflammatory response more effectively resembles the coordinated glial activation reported in the brain tissue of individuals with AD, where microglial and astrocytic responses are geographically and temporally coordinated.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The three-dimensional structure of spheroids provides physiologically realistic oxygen and nutrition gradients that regulate cellular metabolism and inflammatory responses.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The core regions of spheroids experience relative hypoxia compared to peripheral cells, mirroring the oxygen gradients seen in brain tissue (Ausilio et al. 2025).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This hypoxic microenvironment has been observed to enhance inflammatory gene expression through Hypoxia-Inducible Factor (HIF) pathways, leading to elevated production of pro-inflammatory cytokines, such as IL-6 and TNF-α, as evidenced in our experimental findings.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Additionally, the restricted diffusion of nutrients and metabolic waste products in 3D spheroids can induce metabolic stress conditions, which can precondition cells for heightened inflammatory responses.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The metabolic reprogramming that has been observed in this instance is characterized by an increase in glycolytic metabolism.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This metabolic reprogramming has been demonstrated to promote NF-κB activation and inflammatory cytokine production (Poon et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The enhanced inflammatory responses observed in 3D spheroid models provide several advantages for AD research and drug development.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
First, these models better recapitulate the sustained neuroinflammatory state observed in AD patients, where chronic microglial activation and astrogliosis contribute to disease progression The more pronounced inflammatory responses in 3D cultures may provide a more sensitive platform for evaluating anti-inflammatory therapeutic strategies(Park et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Additionally, the enhanced cellular interactions in 3D models enable better assessment of how neuroinflammation affects neuronal networks and synaptic function, which is particularly relevant for understanding the relationship between inflammation and cognitive decline in AD.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Future studies utilizing these enhanced 3D models could provide valuable insights into the temporal dynamics of neuroinflammatory responses and identify critical therapeutic windows for intervention (Park et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
While 3D spheroid models offer significant advantages, it is important to acknowledge their limitations.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The enhanced inflammatory responses may sometimes exceed physiological levels, potentially leading to false positive results in drug screening applications (Gopalakrishnan et al. 2025).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Additionally, the lack of vascular components in current spheroid models limits their ability to fully recapitulate the blood–brain barrier interactions that modulate neuroinflammation in vivo (Cai et al. 2020).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Future developments incorporating microglia-containing organoids and vascularized brain models will further enhance the physiological relevance of these systems.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our results show a significant increase in COX-2 and NOS levels (Fig. 4), which contribute to excessive nitric oxide (NO) production—elevated NO interacts with superoxide radicals, forming peroxynitrite (ONOO −).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These highly reactive nitrogen species cause oxidative damage to cellular components.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This oxidative stress leads to mitochondrial dysfunction and influences epigenetic modifications that suppress BDNF transcription (Retinasamy et al. 2024; Balakrishnan et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Recent findings suggest that oxidative stress-induced DNA hypermethylation of the BDNF promoter region and histone deacetylation reduce gene accessibility, leading to sustained suppression of neurotrophic support (Feng et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Given the established role of mitochondrial dysfunction in AD progression, the interplay between oxidative stress, inflammation, and BDNF downregulation presents a compelling target for neuroprotective interventions (Kouter et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The neuroinflammation caused by LPS is suspected to alter the intracellular signaling pathways of kinases, particularly PKA, AKT, and MAPK, which are implicated in neuronal survival and synaptic integrity (Yin et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Activation of PKA is necessary for the phosphorylation of CREB, which in turn leads to the transcription of BDNF (Liu et al. 2021).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Gene expression analysis revealed significant upregulation of PKA, AKT, and MAPK in SH-SY5Y cells treated with LPS for 24 h (Fig. 5) (Park et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our data suggest that AKT signaling results in reduced mTOR activation, making the cells more susceptible to apoptosis.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The MAPK cascade, particularly the ERK1/2 pathway, is dysfunctional, leading to synaptic anomalies.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
ERK is a vital modulator of memory and synaptic plasticity (Biswas 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The brains of subjects with AD show disruptions in the above signaling pathways, which supports the idea that chronic inflammation leads to this kinase dysfunction, ultimately causing neurodegeneration (Decandia et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The comparison between two types of cells, spheroids and primary neurons, and their neuroinflammatory responses may differ.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our results demonstrate that 3D spheroid cultures of SH-SY5Y cells exhibit a more physiologically relevant inflammatory response to LPS treatment, with significantly higher levels of IL-6 and TNF-α and greater LPS-induced cytotoxicity compared to 2D cultures.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
As per recent reports, 3D neuronal cultures develop more in vivo-like neuronal interactions, thus making it a better model for studying neuroinflammation in AD (Park et al. 2023; Yanakiev et al. 2022).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The more substantial inflammation in 3D cultures is caused by enhanced cell interactions, which are essential components of the inflammatory process in the human brain.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This study has revealed significant differences between 2 and 3D models, suggesting that similar processes may occur in AD and neuroinflammation (Cuni-Lopez et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
With the molecular imbalances documented in this study, various therapeutic strategies warrant investigation to counteract LPS-induced neurotoxicity.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Inhibiting NF-κB (e.g. using BAY 11) or targeting JAK/STAT and NLRP3 pathways could reduce inflammation, offering potential therapeutic strategies for AD (Sivamaruthi et al. 2023; Lee et al. 2012; Xue et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These changes in gene expression and cytokine production provide insights into the molecular mechanisms underlying LPS-induced neuroinflammation and their potential implications for AD pathophysiology.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Antioxidative drugs, such as N-acetylcysteine (NAC) and resveratrol, have shown promise in reducing oxidative stress and restoring BDNF levels in neuroinflammation (Cheng et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Moreover, pharmacological activation of PKA, AKT, and CREB by cAMP analogs or PI3K stimulators is anticipated to reverse the neurotoxic effects of LPS-induced inflammation, thus favoring neuronal survival (Ye et al. 2025; Bhardwaj et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Developments in multi-targeted therapeutic strategies indicate that the simultaneous use of anti-inflammatory agents with neurotrophic enhancers could result in synergistic benefits in treating AD (Cheong et al. 2022; Turgutalp and Kizil 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
With the molecular imbalances documented in this study, various therapeutic strategies are worthy of investigation to reverse LPS-induced neurotoxicity (Colavitta and Barrantes 2023; Nelson et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Despite the significant findings from this research, some limitations must be considered.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
A limitation of this study is that gene expression analysis alone cannot definitively establish functional pathway activation.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Future studies should include protein expression analysis, phosphorylation status assessment, and functional kinase activity measurements to validate these pathways' actual activation status.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
While SH-SY5Y cells are a well-established model for studying neurodegenerative diseases, they fail to capture the complex nature of primary neurons or in vivo brain networks (Lal et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Adding organoid spheroid cultures increases physiological relevance (Hernaiz et al. 2022); To enhance the translational relevance of our findings, future studies should employ in vivo models, such as transgenic AD mouse models (e.g. 5xFAD or APP/PS1), to validate the observed effects of LPS-induced neuroinflammation on BDNF, IL-6, TNF-α, COX-2, NOS, and kinase expression (Cuni-Lopez et al. 2024; Korner et al. 2024).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These models could involve intracerebroventricular or systemic LPS administration to assess neuroinflammatory responses in brain regions relevant to AD, such as the hippocampus and cortex, and evaluate the efficacy of inhibitors targeting NF-κB, JAK/STAT, or NLRP3 pathways (Zhao et al. 2019; Varma et al. 2023).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Such studies would bridge the gap between our in vitro findings and clinical applications, providing a more comprehensive understanding of neuroinflammatory mechanisms in AD.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
In conclusion, our findings demonstrate that LPS-induced neuroinflammation in SH-SY5Y cells leads to reduced cell viability (Fig. 1), decreased BDNF levels (Fig. 2), increased pro-inflammatory markers (Figs. 3 and 4), and upregulated expression of key signaling kinases (Fig. 5).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These results collectively highlight the complex interplay between inflammation, neurotrophic support, and signaling pathways in AD, suggesting potential therapeutic targets for mitigating neuroinflammatory damage.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Neuroinflammation plays an important role in AD progression and is responsible for synaptic dysfunction, neuronal death, and cognitive decline.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our research shows that exposure to LPS-induced neuroinflammation affects neuronal viability, neurotrophic signaling, and inflammatory marker expression in SH-SY5Y cells, primary neurons, and 3D spheroids.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
The downregulation of BDNF and upregulation of PKA, AKT, and MAPK gene expression in response to LPS-induced neuroinflammation are associated with impaired neuronal survival in SH-SY5Y cells.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
We demonstrate that 3D spheroid cultures exhibit a more physiological inflammatory response compared to 2D SH-SY5Y cultures.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This implies that 3D setups may offer a more precise platform for drug discovery and therapeutic evaluation in AD studies.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Furthermore, our results suggest that NF-κB-driven inflammation, potentially linked to increased PKA expression and elevated IL-6 and TNF-α levels,contributes to neuronal dysfunction; however, future studies should directly monitor NF-κB activation to confirm this involvement.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Additionally, targeting NF-κB, oxidative stress, or kinase pathways may offer viable therapeutic strategies to mitigate inflammation-induced neurodegeneration.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
According to the above information, selecting a suitable model or parameter is crucial for better studying diseases like AD.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These results collectively underscore the complex interplay between inflammation, neurotrophic support, and signaling pathways in AD, suggesting potential therapeutic targets for mitigating inflammatory damage in neurodegenerative diseases.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Background/Objectives: Satoyoshi syndrome is a rare, autoimmune disorder currently diagnosed based on clinical criteria: painful muscle spasms, diarrhea, and alopecia.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Two previous reports showed a specific immunoreactive band in three Satoyoshi syndrome patients using Western blot analysis, with brain homogenate as the antigen source.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
These findings could be the basis for a future diagnostic test.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The aim of our study was to evaluate the efficacy of using SH-SY5Y cell lysate instead of brain homogenate for a potential laboratory test for Satoyoshi syndrome using the Western blot technique.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Methods: Western blot analyses were conducted using brain homogenate, SH-SY5Y cell lysates, and differentiated SH-SY5Y cell lysates.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Serum samples were obtained from three Satoyoshi syndrome patients, alongside control samples from thirty blood donors and six patients with other neurological conditions.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Results: Sera from patients with Satoyoshi syndrome displayed a three-band pattern in the 70–100 kDa range.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
This pattern was reproducible across all tested antigen sources (brain homogenate, SH-SY5Y lysate, and differentiated SH-SY5Y lysate) but was not observed for the sera from the control groups.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The bands were more visible when using either type of SH-SY5Y lysate compared to brain homogenate.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
No differences were found between the SH-SY5Y lysate and the differentiated SH-SY5Y lysate.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Conclusions: Sera from our Satoyoshi syndrome patients showed a specific band pattern that could be used for a future evaluation of Satoyoshi syndrome using Western blot.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The use of SH-SY5Y cell lysate vs. brain homogenate as an antigen source may improve visualization and reproducibility of the immunobands and be less costly.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Satoyoshi syndrome (OMIM: 600705) is a rare, multisystemic disorder characterized by a spectrum of clinical features, including painful intermittent muscle spasms, diarrhea, alopecia, skeletal abnormalities, growth retardation, and endocrinopathies .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
A delay in diagnosis and the subsequent lack of appropriate treatment can lead to poor outcomes, including severe complications and potentially fatal progression .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
It is believed that Satoyoshi syndrome is an autoimmune disease.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The association betweenSatoyoshi syndrome andother autoimmune diseases (thyroiditis, myasthenia, systemic lupus, atopic dermatitis, and idiopathic thrombocytopenic purpura), the presence of autoantibodies (ANAs, anti-acetylcholine receptor (AChR), anti-glutamic acid decarboxylase, among others), and the improvement insymptoms with immune response modifiers are considered evidence supporting an autoimmune origin .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The abovementioned serum autoantibodies are not specific to Satoyoshi syndrome.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
However, an antibody against brain homogenate, which was not found in healthy controls, has been reported in three previous cases using Western blot.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Endo et al., 2001 , described a 13-year-old female patient with Satoyoshi syndrome who carried the anti-AChR autoantibody, without presenting symptoms of myasthenia gravis.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Analysis of this patient’s serum using Western blot on brain homogenate revealed an 85 kDa immunoreactive band, although the specific antigen remained unidentified .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
Similarly, Matsuura et al., 2007, reported two female patients (aged 17 and 36) with classic symptoms of Satoyoshi syndrome, whose sera showed a 90 kDa band when tested with brain, stomach, and duodenum homogenates using Western blot .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
The aim of this study was to investigate whether the sera from three otherSatoyoshi patients from Europe also carry specific autoantibodies as assessed by Western blot, using either brain homogenate or SH-SY5Y cell lysate as the antigen source.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
SH-SY5Y is a well-known human neuroblastoma cell line derived from a bone marrow biopsy of a patient with neuroblastoma.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
It is widely used in neuroscience research as an in vitro model to study neuronal differentiation, function, and processes related to neurodegenerative diseases, like Parkinson’s disease .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
This cell line was chosen for this study as an alternative to human brain homogenate because it is well described in the literature, easy to obtain commercially, and, therefore, facilitates the standardization of the procedure.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
In a first experiment, we assessed the autoimmune immunoreactivity of the serum from three Satoyoshi patients on brain homogenate and two types of SH-SY5Y cell lysates, namely, standard and after inducing neuronal differentiation, to determine which source of antigen would be more suitable.
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
SH-SY5Y (Cellosaurus database accession number: CVCL_0019); https://www.cellosaurus.org/CVCL_0019 (accessed on 15 September 2025) is a neuroblastoma cell line commonly used as an in vitro model for neurological disease .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
These cells are susceptible to being differentiated into mature neurons .
PMC12610766
Preliminary Evidence for a Western Blot Diagnosis of Satoyoshi Syndrome Using SH-SY5Y Neuroblastoma Cell Lysate as the Antigen Source
SH-SY5Y cells were purchased from the American Type Culture Collection, https://www.atcc.org/. In a second experiment, we aimed to compare the immunobands obtained from the serum samples of Satoyoshi patients using Western blot with the immunobands from the sera of healthy blood donors and patients with other neurological diseases to investigate the possibility of a specific band pattern for Satoyoshi patients.