Abstract
The anti-inflammatory and neuroprotective effects of short chain fatty acid (SCFA) butyrate have been explored in a wide array of neurological pathologies. It is a 4-carbon SCFA produced from the fermentation of dietary fibers by the gut-microbiota. As evident from previous literature, butyrate plays a wide array of functions in CNS and interestingly enhances the differentiation potential of Neural stem/Progenitor Cells (NSPCs). Japanese encephalitis virus (JEV) is a well-known member of the Flaviviridae family and has been shown to alter neural stem cell pool of the brain, causing devastating consequences. In this study, we administered sodium butyrate (NaB) post JEV infection in BALB/c mouse model to examine any possible amelioration of the viral infection in NSPCs. In addition, ex vivo neurospheres and in vitro model of NSPCs were also used to study the effect of sodium butyrate in JEV infection. As an unprecedented finding, butyrate treated infected animals presented early onset of symptoms, as compared to their respective JEV infected groups. Alongside, we observed an increased viral load in NSPCs isolated from these animals as well as in cell culture models upon sodium butyrate treatment. Cytometric bead array analysis also revealed an increase in inflammatory cytokines, particularly, MCP-1 and IL-6. Further, increased expression of the key members of the canonical NF-κB pathway, viz-a-viz p-NF-κB, p-Iκ-Bα and p-IKK was observed. Overall, the increased inflammation and cell death caused early symptom progression in NaB-treated JEV infected animal model, which is contradictory to the well documented protective nature of NaB and therefore a better understanding of SCFA-based modulation of the gut-brain axis in viral infections is required.
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Abbreviations
- CNS:
-
Central nervous system
- BBB:
-
Blood brain barrier
- SCFA:
-
short chain fatty acid
- HDAC:
-
histone deacetylase
- NSC:
-
neural stem cells
- NSPC:
-
neural stem/ progenitor cell
- SGZ:
-
sub granular zone
- SVZ:
-
sub-ventricular zone
- JEV:
-
Japanese encephalitis virus
- NaB:
-
sodium butyrate
- MEM:
-
minimum essential media
- MOI:
-
multiplicity of infection
- CBA:
-
cytometric bead array
- HPI:
-
hours post infection
- TNF-α:
-
Tumor necrosis factor-alpha
- IL-6:
-
Interleukin-6
- MCP-1:
-
Monocyte chemoattractant protein-1
- NS3:
-
Nonstructural protein 3
- NF-κB:
-
Nuclear factor- kappa B
- IKβ:
-
inhibitor of NF-κB
- IKK:
-
IKβ kinase
- PBS:
-
Phosphate buffered saline
- PFU/ml:
-
plaque forming unit per milliiter
- DMSO:
-
Dimethyl sulfoxide
- RIG-1:
-
Retinoic acid inducible gene -1
- GAPDH:
-
Glyceraldehyde phosphate-3 dehydrogenese
- FBS:
-
Fetal bovine serum
- DMEM:
-
Dulbecco’s Modified Eagle Medium
- DMEM/F-12:
-
Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12
- BCA:
-
Bicinchonic acid
- SDS- PAGE:
-
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- HRP:
-
Horse radish peroxidase
- IL-12:
-
interleukin 12
- IL-10:
-
interleukin 10
- IFN-γ:
-
Interferon gamma
- MTS:
-
3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
- DG:
-
Dentate gyrus
- ZIKV:
-
Zika virus
- HIV:
-
Human immunodeficiency virus
- VSV:
-
Vesicular Stomatitis Virus
- SARS-CoV-2:
-
Severe acute respiratory syndrome coronavirus 2
- EBV:
-
Epstein Barr Virus
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Acknowledgements
We would like to sincerely acknowledge Dr. Atreye Majumdar for reviewing and providing her critical suggestions in developing the manuscript; Stuti Mohapatra and Indira Priya Siva Venkatesh for their contributions in critically reviewing the manuscript. Additionally, we acknowledge Mr. Manish Kumar Dogra and Dr. Kanhaiya Lal Kumawat for their technical assistance.
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This work was supported by core grant to AB from National Brain Research Centre. AB is also a recipient of J C Bose Fellowship (JCB/2020/000037) from the Science and Engineering Research Board (SERB), Ministry of Science and Technology, Government of India.
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Anirban Basu and Shivangi Sharma contributed to the study conception and design. Material preparation, data collection and analysis were performed by Anirudh Satheesan and Shivangi Sharma. The manuscript was written by Anirudh Satheesan and Shivangi Sharma. Anirban Basu critically reviewed the manuscript.
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Satheesan, A., Sharma, S. & Basu, A. Sodium Butyrate Induced Neural Stem/Progenitor Cell Death in an Experimental Model of Japanese Encephalitis. Metab Brain Dis 38, 2831–2847 (2023). https://doi.org/10.1007/s11011-023-01279-3
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DOI: https://doi.org/10.1007/s11011-023-01279-3