GO:0043524 negative regulation of neuron apoptotic process: Neuroprotection Pathway, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0043524 describes any biological process that stops, prevents, or reduces apoptotic death of neurons, thereby supporting neuronal survival.
• Key molecular effectors include anti-apoptotic signaling through PINK1/Parkin, MAPK pathway modulation, and microRNA-mediated control of pro-apoptotic transcripts.
• Dysregulation of this process contributes to ischemic stroke, Alzheimer's disease, seizure-induced brain injury, and anesthesia-related neurotoxicity.
• MicroRNAs such as miR-30c-5p, miR-125a, miR-181a-5p, miR-23a-3p, and miR-223 are critical regulators of neuronal apoptosis.
• Experimental models for studying GO:0043524 include oxygen-glucose deprivation, H2O2-induced oxidative stress, sevoflurane exposure, febrile seizure, and amyloid-beta toxicity.
• CRISPR-based knockout, knock-in, and overexpression models enable causal testing of candidate neuroprotective genes in neuronal cell lines and primary neurons.
Description
The Gene Ontology term GO:0043524, negative regulation of neuron apoptotic process, defines any process that stops, prevents, or reduces the frequency, rate, or extent of apoptotic cell death specifically in neurons. Apoptosis is a programmed cell death mechanism that is essential for normal nervous system development but becomes pathological when it eliminates neurons in neurodegenerative diseases, stroke, or trauma. Understanding how neurons resist apoptosis is therefore central to neurobiology and to the development of neuroprotective therapies. This GO term captures a wide range of molecular events, including activation of survival kinases, inhibition of pro-apoptotic Bcl-2 family members, regulation by microRNAs, and modulation of mitochondrial quality control pathways. Research into GO:0043524 has revealed that neuroprotection is often mediated by signaling cascades such as PINK1/Parkin-dependent mitophagy, MAPK pathway suppression, and Wnt/beta-catenin signaling. These pathways converge on the core apoptotic machinery to prevent cytochrome c release, caspase activation, and DNA fragmentation in neurons. Because neuronal apoptosis is a final common pathway in many neurological disorders, the negative regulation of this process represents a high-value target for both mechanistic studies and therapeutic intervention.
negative regulation of neuron apoptotic process At A Glance
| GO ID | GO:0043524 |
|---|---|
| GO term | negative regulation of neuron apoptotic process |
| Ontology | biological_process |
| Definition | Any process that stops, prevents, or reduces the frequency, rate or extent of cell death by apoptotic process in neurons. |
| Synonyms | down regulation of neuron apoptosis; down-regulation of neuron apoptosis; downregulation of neuron apoptosis; inhibition of neuron apoptosis; negative regulation of neuron apoptosis; negative regulation of programmed cell death, neurons; neuron survival |
| Major function | Promotes neuronal survival by inhibiting apoptotic cell death pathways |
| Related processes | Mitophagy, MAPK signaling, Wnt/beta-catenin signaling, microRNA regulation, caspase inhibition |
| Disease relevance | Ischemic stroke, Alzheimer's disease, seizure-induced brain injury, anesthesia neurotoxicity |
What Is GO:0043524?
GO:0043524, negative regulation of neuron apoptotic process, is defined as any process that stops, prevents, or reduces the frequency, rate or extent of cell death by apoptotic process in neurons. In simpler terms, it encompasses all biological mechanisms that keep neurons alive by blocking the apoptotic program. This includes extracellular survival signals, intracellular anti-apoptotic proteins, microRNA-mediated repression of pro-apoptotic genes, and mitochondrial protective pathways. The term is a biological process and is distinct from general negative regulation of apoptosis because it is specific to neurons.
Why Is negative regulation of neuron apoptotic process Important in Cell Biology?
GO:0043524 is critically important because neuronal apoptosis underlies the pathogenesis of many acute and chronic neurological disorders, including ischemic stroke, Alzheimer's disease, epilepsy-associated brain injury, and anesthetic neurotoxicity. Identifying the molecular players that negatively regulate this process can reveal therapeutic targets for neuroprotection. Moreover, the term provides a standardized framework for annotating gene function in neuroscience, enabling researchers to compare results across studies and model systems.
• Neuronal apoptosis is a major contributor to ischemic stroke injury, and enhancing its negative regulation reduces infarct volume.
• In Alzheimer's disease, dysregulated neuronal apoptosis correlates with cognitive decline, making its inhibition a therapeutic goal.
• Febrile seizures can trigger hippocampal neuron apoptosis, and microRNA-223 upregulation protects against this injury.
• Sevoflurane anesthesia can induce neuron injury, and miR-181a-5p knockdown ameliorates this via DDX3X/Wnt/beta-catenin signaling.
• Irradiation-induced neuronal apoptosis is mediated by downregulation of miR-23a-3p, highlighting microRNA control of this process.
• Oxidative stress in PC-12 cells is a common model for studying negative regulation of neuronal apoptosis.
• The PINK1/Parkin pathway protects neurons against oxidative stress by promoting mitophagy and reducing apoptosis.
• Long non-coding RNAs such as PVT1 and ANRIL modulate neuronal apoptosis through microRNA sponging and MAPK signaling.
• Understanding GO:0043524 aids in developing neuroprotective strategies for neurodegenerative diseases and acute brain injuries.
• CRISPR-based models allow causal testing of candidate genes in the negative regulation of neuron apoptotic process.
What Happens During negative regulation of neuron apoptotic process?
Initiation of Survival Signaling
In simple terms: Neurons receive signals that tell them to stay alive.
Negative regulation of neuron apoptotic process begins with extracellular or intracellular signals that activate pro-survival pathways. For example, human neural stem cell-derived exosomes activate the PINK1/Parkin pathway to protect against oxidative stress-induced neuronal injury in ischemic stroke. Similarly, upregulation of microRNA-223 inhibits brain injury and hippocampal neuron apoptosis after febrile seizure through the NLRP3-Caspase-1 signaling pathway. These initiating events set in motion a cascade that blocks the apoptotic machinery.
Mitochondrial Protection and Mitophagy
In simple terms: The cell cleans up damaged mitochondria to prevent them from triggering death.
Mitochondria are central to apoptosis. Negative regulation of neuronal apoptosis often involves mitophagy, the selective removal of damaged mitochondria. Activation of PINK1/Parkin by exosomes from neural stem cells protects neurons against oxidative stress by enhancing mitophagy and reducing apoptotic markers. This mitochondrial quality control prevents the release of cytochrome c and subsequent caspase activation, thereby promoting neuronal survival.
MicroRNA-Mediated Repression of Pro-Apoptotic Genes
In simple terms: Small RNA molecules put brakes on genes that would otherwise kill the neuron.
MicroRNAs are key regulators of neuronal apoptosis. miR-30c-5p is inhibited by the long non-coding RNA PVT1, which upregulates Rock2 and modulates cerebral ischemia/reperfusion injury through MAPK signaling. Knockdown of ANRIL aggravates H2O2-induced injury in PC-12 cells by targeting microRNA-125a. miR-181a-5p knockdown ameliorates sevoflurane anesthesia-induced neuron injury via regulation of the DDX3X/Wnt/beta-catenin signaling axis. Downregulation of miR-23a-3p mediates irradiation-induced neuronal apoptosis. Upregulation of microRNA-223 inhibits hippocampal neuron apoptosis through the NLRP3-Caspase-1 pathway. These examples illustrate how microRNAs fine-tune the apoptotic threshold in neurons.
Inhibition of Caspase Activation and Apoptotic Execution
In simple terms: The final executioner proteins that dismantle the cell are blocked.
The ultimate step in negative regulation of neuron apoptotic process is the inhibition of caspases, the proteases that execute apoptosis. For instance, miR-223 upregulation inhibits the NLRP3-Caspase-1 signaling pathway, reducing caspase-1 activity and neuronal apoptosis after febrile seizure. Similarly, SIRT4 promotes neuronal apoptosis in Alzheimer's disease models via the STAT2-SIRT4-mTOR pathway, indicating that mTOR signaling can modulate apoptotic execution. By preventing caspase activation, neurons avoid DNA fragmentation and cell death.
Modulation by Long Non-Coding RNAs and Signaling Pathways
In simple terms: Other regulatory molecules and pathways help decide whether the neuron lives or dies.
Long non-coding RNAs (lncRNAs) such as PVT1 and ANRIL modulate neuronal apoptosis by sponging microRNAs and affecting downstream signaling. PVT1 inhibits miR-30c-5p to upregulate Rock2, activating MAPK signaling in cerebral ischemia/reperfusion injury. ANRIL knockdown aggravates H2O2-induced injury in PC-12 cells by targeting microRNA-125a. These lncRNAs act as molecular switches that can tip the balance toward survival or death, and their dysregulation contributes to neurological disorders.
Key Genes Involved in GO:0043524 negative regulation of neuron apoptotic process
The following genes and non-coding RNAs have been experimentally implicated in the negative regulation of neuron apoptotic process, based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| PINK1 | Activates mitophagy to protect neurons from oxidative stress | Studied in ischemic stroke models using exosome treatment |
| Parkin | E3 ubiquitin ligase that works with PINK1 in mitophagy | Key effector in neuroprotection against oxidative stress |
| PVT1 | Long non-coding RNA that sponges miR-30c-5p | Modulates cerebral ischemia/reperfusion injury via MAPK signaling |
| miR-30c-5p | MicroRNA that targets Rock2 | Inhibited by PVT1; affects neuronal apoptosis in ischemia |
| Rock2 | Rho-associated kinase 2, pro-apoptotic when upregulated | Upregulated by PVT1/miR-30c-5p axis in ischemia |
| ANRIL | Long non-coding RNA that regulates miR-125a | Knockdown aggravates H2O2-induced injury in PC-12 cells |
| miR-125a | MicroRNA targeted by ANRIL | Protects PC-12 cells from oxidative stress-induced apoptosis |
| miR-181a-5p | MicroRNA that modulates DDX3X/Wnt/beta-catenin signaling | Knockdown ameliorates sevoflurane-induced neuron injury |
| DDX3X | RNA helicase involved in Wnt/beta-catenin signaling | Regulated by miR-181a-5p in anesthesia neurotoxicity |
| SIRT4 | Mitochondrial sirtuin that promotes apoptosis | Promotes neuronal apoptosis in Alzheimer's disease via STAT2-SIRT4-mTOR |
| STAT2 | Transcription factor that regulates SIRT4 | Part of the STAT2-SIRT4-mTOR pathway in Alzheimer's disease |
| mTOR | Kinase that regulates cell survival and apoptosis | Modulated by SIRT4 in Alzheimer's disease models |
| miR-23a-3p | MicroRNA downregulated by irradiation | Mediates irradiation-induced neuronal apoptosis |
| miR-223 | MicroRNA that inhibits NLRP3-Caspase-1 pathway | Upregulation inhibits hippocampal neuron apoptosis after febrile seizure |
| NLRP3 | Inflammasome component that activates caspase-1 | Inhibited by miR-223 to reduce neuronal apoptosis |
| Caspase-1 | Protease that executes apoptosis | Inhibited by miR-223 in febrile seizure model |
How Is negative regulation of neuron apoptotic process Regulated?
The negative regulation of neuron apoptotic process is controlled by multiple layers of regulation. At the post-transcriptional level, microRNAs such as miR-30c-5p, miR-125a, miR-181a-5p, miR-23a-3p, and miR-223 directly repress pro-apoptotic targets or modulate signaling pathways. Long non-coding RNAs like PVT1 and ANRIL act as competing endogenous RNAs to sponge these microRNAs, thereby influencing neuronal survival. At the signaling level, the PINK1/Parkin pathway promotes mitophagy and protects against oxidative stress, while the MAPK pathway can either promote or inhibit apoptosis depending on context. The Wnt/beta-catenin signaling axis is modulated by miR-181a-5p and DDX3X in anesthesia-induced neurotoxicity. Additionally, the STAT2-SIRT4-mTOR pathway regulates neuronal apoptosis in Alzheimer's disease models. These regulatory mechanisms collectively determine the threshold for apoptosis in neurons.
negative regulation of neuron apoptotic process and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PINK1/Parkin | Ischemic stroke | Oxygen-glucose deprivation in primary neurons or SH-SY5Y cells |
| PVT1/miR-30c-5p/Rock2 | Cerebral ischemia/reperfusion injury | Middle cerebral artery occlusion in rats or OGD in PC-12 cells |
| ANRIL/miR-125a | Oxidative stress-induced neuronal injury | H2O2 treatment in PC-12 cells |
| miR-181a-5p/DDX3X | Sevoflurane anesthesia-induced neurotoxicity | Sevoflurane exposure in primary neurons or neuronal cell lines |
| SIRT4/STAT2/mTOR | Alzheimer's disease | Amyloid-beta treatment in primary neurons or APP/PS1 mice |
| miR-23a-3p | Irradiation-induced neuronal apoptosis | Irradiation of hippocampal neurons or neuronal progenitors |
| miR-223/NLRP3/Caspase-1 | Febrile seizure | Hyperthermia-induced seizure in rat pups |
Ischemic Stroke
Ischemic stroke causes oxidative stress and neuronal apoptosis. Human neural stem cell-derived exosomes activate the PINK1/Parkin pathway to protect against oxidative stress-induced neuronal injury, demonstrating that enhancing negative regulation of neuron apoptotic process is neuroprotective. The long non-coding RNA PVT1 modulates cerebral ischemia/reperfusion injury through the miR-30c-5p/Rock2/MAPK axis, further highlighting the role of this process in stroke pathology.
Alzheimer's Disease
In Alzheimer's disease, neuronal apoptosis contributes to neurodegeneration. SIRT4 promotes neuronal apoptosis via the STAT2-SIRT4-mTOR pathway, suggesting that inhibiting SIRT4 or activating mTOR could be protective. The negative regulation of neuron apoptotic process is therefore a potential therapeutic target in Alzheimer's disease.
Seizure-Induced Brain Injury
Febrile seizures can lead to hippocampal neuron apoptosis. Upregulation of microRNA-223 inhibits brain injury and hippocampal neuron apoptosis through the NLRP3-Caspase-1 signaling pathway, indicating that microRNA-based interventions can enhance negative regulation of neuron apoptotic process.
Anesthesia-Induced Neurotoxicity
Sevoflurane anesthesia can induce neuron injury. MiR-181a-5p knockdown ameliorates sevoflurane anesthesia-induced neuron injury via regulation of the DDX3X/Wnt/beta-catenin signaling axis, showing that targeting this microRNA can promote neuronal survival. Irradiation-induced neuronal apoptosis is mediated by downregulation of miR-23a-3p, further linking microRNA dysregulation to neuronal death.
From negative regulation of neuron apoptotic process-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does knockout of a candidate gene enhance or reduce neuronal apoptosis? | CRISPR knockout in neuronal cell lines (e.g., SH-SY5Y, PC-12) or primary neurons |
| Does a specific point mutation in an anti-apoptotic gene alter its protective function? | CRISPR point mutation knock-in in neuronal cells |
| Does knock-in of a tagged survival protein affect its localization during apoptosis? | CRISPR knock-in of fluorescent or epitope tags |
| Does overexpression of a microRNA or lncRNA protect neurons from oxidative stress? | Lentiviral overexpression in primary neurons or cell lines |
| Does knockdown of a pro-apoptotic microRNA ameliorate anesthesia-induced injury? | CRISPR interference or antagomir treatment in neuronal cultures |
| Does activation of PINK1/Parkin mitigate ischemic neuronal death? | Exosome treatment or small-molecule activators in OGD models |
How to Study the negative regulation of neuron apoptotic process Process
| Method | What It Measures | Typical Application |
|---|---|---|
| TUNEL assay | DNA fragmentation indicative of apoptosis | Quantifying neuronal death in stroke or seizure models |
| Caspase-3/7 activity assay | Executioner caspase activity | Assessing apoptosis in H2O2-treated PC-12 cells |
| Annexin V flow cytometry | Phosphatidylserine externalization | Detecting early apoptosis in neuronal cultures |
| Mitochondrial membrane potential assay | Mitochondrial integrity | Evaluating PINK1/Parkin-mediated protection |
| Western blot | Protein expression and cleavage | Measuring cleaved caspase-3, PARP, or Bcl-2 family proteins |
| qRT-PCR | MicroRNA and mRNA levels | Validating microRNA targets in neuronal apoptosis |
| Luciferase reporter assay | MicroRNA-target interaction | Confirming miR-30c-5p binding to Rock2 |
| Immunofluorescence | Protein localization and apoptosis markers | Visualizing cytochrome c release or nuclear condensation |
CRISPR Knockout and Knock-in Models
CRISPR/Cas9 technology enables precise knockout or knock-in of genes involved in negative regulation of neuron apoptotic process. For example, knocking out PINK1 or Parkin can test their requirement for exosome-mediated neuroprotection. Point mutations can be introduced to dissect domain-specific functions of anti-apoptotic proteins. These models provide causal evidence linking specific genes to neuronal survival.
MicroRNA and lncRNA Functional Studies
MicroRNAs and long non-coding RNAs are key regulators of neuronal apoptosis. Methods include microRNA mimics, inhibitors (antagomirs), and lncRNA overexpression or knockdown. For instance, miR-223 upregulation inhibits hippocampal neuron apoptosis through the NLRP3-Caspase-1 pathway, while ANRIL knockdown aggravates H2O2-induced injury by targeting miR-125a. These approaches help delineate the regulatory networks controlling GO:0043524.
Apoptosis Assays and Imaging
To measure negative regulation of neuron apoptotic process, researchers use TUNEL staining, caspase-3/7 activity assays, Annexin V flow cytometry, and mitochondrial membrane potential measurements. Imaging of cytochrome c release and nuclear condensation provides spatial information. These assays are applied in models such as oxygen-glucose deprivation, H2O2 treatment, sevoflurane exposure, and febrile seizure.
Transcriptomics and Proteomics
RNA sequencing and proteomics can identify global changes in gene expression during neuronal apoptosis. For example, transcriptomic analysis after PVT1 modulation revealed changes in MAPK signaling components. Proteomic profiling of mitophagy-related proteins can uncover PINK1/Parkin substrates. These unbiased approaches generate hypotheses about novel regulators of GO:0043524.
How CRISPR Can Be Used to Study GO:0043524 negative regulation of neuron apoptotic process
Knockout
CRISPR knockout of genes such as PINK1, Parkin, or SIRT4 can determine whether they are required for negative regulation of neuron apoptotic process. For example, knocking out PINK1 would test its necessity in exosome-mediated neuroprotection against oxidative stress. Knockout of SIRT4 might reduce neuronal apoptosis in Alzheimer's disease models. These experiments provide loss-of-function evidence.
Point Mutation
Point mutations can be introduced to dissect specific residues or domains critical for anti-apoptotic function. For instance, mutating phosphorylation sites in PINK1 or Parkin could reveal their role in mitophagy and neuroprotection. Similarly, point mutations in DDX3X could clarify its regulation by miR-181a-5p in anesthesia-induced neurotoxicity.
Knock-in
Knock-in of tagged versions of survival proteins (e.g., GFP-Parkin) allows real-time imaging of their localization during apoptosis. Knock-in of disease-associated mutations, such as those in SIRT4 or mTOR, can model Alzheimer's disease-related neuronal apoptosis. These models are valuable for studying dynamic processes in live neurons.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression can upregulate protective genes or microRNAs. Overexpression of miR-223 inhibits hippocampal neuron apoptosis after febrile seizure, while overexpression of PINK1/Parkin enhances neuroprotection. These gain-of-function studies complement knockout approaches.
How EDITGENE Supports negative regulation of neuron apoptotic process Research
Researchers studying negative regulation of neuron apoptotic process-related genes often need to determine whether a candidate gene is causally involved in neuronal survival or death. EDITGENE provides comprehensive CRISPR-based services to accelerate this research, from knockout and point mutation to knock-in and overexpression models, as well as library screening and bioinformatics support.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of neuron apoptotic process research.
Frequently Asked Questions About negative regulation of neuron apoptotic process
What is GO:0043524?
GO:0043524 is the Gene Ontology term for negative regulation of neuron apoptotic process, defined as any process that stops, prevents, or reduces the frequency, rate or extent of apoptotic cell death in neurons.
What genes are involved in negative regulation of neuron apoptotic process?
Key genes include PINK1, Parkin, SIRT4, STAT2, mTOR, DDX3X, and non-coding RNAs such as PVT1, ANRIL, miR-30c-5p, miR-125a, miR-181a-5p, miR-23a-3p, and miR-223.
How is neuronal apoptosis inhibited?
Neuronal apoptosis is inhibited by activation of survival signaling pathways (e.g., PINK1/Parkin, Wnt/beta-catenin), microRNA-mediated repression of pro-apoptotic genes, and blockade of caspase activation.
What diseases involve dysregulated neuron apoptotic process?
Ischemic stroke, Alzheimer's disease, seizure-induced brain injury, and anesthesia-induced neurotoxicity all involve dysregulation of neuronal apoptosis.
What experimental models are used to study GO:0043524?
Common models include oxygen-glucose deprivation, H2O2-induced oxidative stress in PC-12 cells, sevoflurane exposure, febrile seizure in rats, and amyloid-beta treatment in neurons.
How do microRNAs regulate neuronal apoptosis?
MicroRNAs such as miR-223, miR-181a-5p, and miR-23a-3p directly repress pro-apoptotic targets or modulate signaling pathways to influence neuronal survival.
What is the role of PINK1/Parkin in neuroprotection?
PINK1 and Parkin promote mitophagy, removing damaged mitochondria and preventing oxidative stress-induced neuronal apoptosis.
Can CRISPR be used to study negative regulation of neuron apoptotic process?
Yes, CRISPR knockout, knock-in, point mutation, and overexpression models enable causal testing of genes involved in neuronal survival.
What is the role of lncRNAs in neuronal apoptosis?
Long non-coding RNAs such as PVT1 and ANRIL act as microRNA sponges to modulate apoptotic signaling in neurons.
How does SIRT4 affect neuronal apoptosis in Alzheimer's disease?
SIRT4 promotes neuronal apoptosis via the STAT2-SIRT4-mTOR pathway, suggesting that inhibiting SIRT4 could be protective.
Conclusion
GO:0043524, negative regulation of neuron apoptotic process, is a fundamental biological process that protects neurons from programmed cell death. Research has identified diverse molecular players, including PINK1/Parkin, microRNAs, long non-coding RNAs, and signaling pathways such as MAPK and Wnt/beta-catenin, that collectively maintain neuronal survival. Dysregulation of this process contributes to ischemic stroke, Alzheimer's disease, seizure-induced injury, and anesthesia neurotoxicity, making it a prime target for therapeutic intervention. CRISPR-based models and advanced omics technologies continue to unravel the complex regulatory networks underlying this process, offering hope for new neuroprotective strategies.
References
- 1. Zhao M et al.. 2025. Human neural stem cell-derived exosomes activate PINK1/Parkin pathway to protect against oxidative stress-induced neuronal injury in ischemic stroke.. J Transl Med 23(1):402 PMID: 40188077
- 2. Zhang H et al.. 2021. Long Non-coding RNA PVT1 Inhibits miR-30c-5p to Upregulate Rock2 to Modulate Cerebral Ischemia/Reperfusion Injury Through MAPK Signaling Pathway Activation.. Mol Neurobiol 58(11):6032-6048 PMID: 34436749
- 3. Li R et al.. 2017. Knockdown of ANRIL aggravates H(2)O(2)-induced injury in PC-12 cells by targeting microRNA-125a.. Biomed Pharmacother 92:952-961 PMID: 28609843
- 4. She Y et al.. 2024. MiR-181a-5p knockdown ameliorates sevoflurane anesthesia-induced neuron injury via regulation of the DDX3X/Wnt/β-catenin signaling axis.. Exp Brain Res 242(3):571-583 PMID: 38218948
- 6. Xing D et al.. 2024. SIRT4 promotes neuronal apoptosis in models of Alzheimer's disease via the STAT2-SIRT4-mTOR pathway.. Am J Physiol Cell Physiol 326(6):C1697-C1709 PMID: 38586875
- 7. Sabirzhanov B et al.. 2020. Down-Regulation of miR-23a-3p Mediates Irradiation-Induced Neuronal Apoptosis.. Int J Mol Sci 21(10) PMID: 32456284
- 8. Wang B et al.. 2019. Up-regulation of microRNA-223 inhibits brain injury and hippocampal neuron apoptosis of rats after febrile seizure through the NLRP3-Caspase-1 signaling pathway.. Biomed Pharmacother 114:108683 PMID: 30947016
