Abstract
Hepatic encephalopathy (HE) is a debilitating neurological disorder associated with liver failure and characterized by impaired brain function. Decade-long studies have led to significant advances in our understanding of HE; however, effective therapeutic management of HE is lacking, and HE continues to be a significant cause of morbidity and mortality in patients, underscoring the need for continued research into its pathophysiology and treatment. Accordingly, the present study provides a comprehensive overview aimed at elucidating the molecular underpinnings of HE and identifying potential therapeutic targets. A moderate-grade HE model was induced in rats using thioacetamide, which simulates the liver damage observed in patients, and its impact on cognitive function, neuronal arborization, and cellular morphology was also evaluated. We employed label-free LC–MS/MS proteomics to quantitatively profile hippocampal proteins to explore the molecular mechanism of HE pathogenesis; 2175 proteins were identified, 47 of which exhibited significant alterations in moderate-grade HE. The expression of several significantly upregulated proteins, such as FAK1, CD9 and Tspan2, was further validated at the transcript and protein levels, confirming the mass spectrometry results. These proteins have not been previously reported in HE. Utilizing Metascape, a tool for gene annotation and analysis, we further studied the biological pathways integral to brain function, including gliogenesis, the role of erythrocytes in maintaining blood–brain barrier integrity, the modulation of chemical synaptic transmission, astrocyte differentiation, the regulation of organ growth, the response to cAMP, myelination, and synaptic function, which were disrupted during HE. The STRING database further elucidated the protein‒protein interaction patterns among the differentially expressed proteins. This study provides novel insights into the molecular mechanisms driving HE and paves the way for identifying novel therapeutic targets for improved disease management.
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Data availability
The raw mass spectrometry data are available from the ProteomeXchange consortium and can be accessed via the dataset identifier PXD036723 (Username: reviewer_pxd036723@ebi.ac.uk Password: Jcvbq2v0). Supplementary figures and tables can be found in the shared supplementary files. Other data related to this work can be shared upon reasonable request to the corresponding author.
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Acknowledgements
The authors acknowledge Prof. SK Trigun Laboratory facilities, Department of Zoology, BHU, Prof Sanjeeva Srivastava, BSBE, and IIT Bombay for the MASSFIITB facility (BT/PR13114/INF/22/206/2015), the Central Discovery Centre, BHU for Trinocular Research Microscope and the Interdisciplinary School of Life Sciences, BHU for the qPCR facility. SKP thanks the Science and Engineering Research Board, Government of India, for the project assistance fellowship and the Indian Council of Medical Research for a senior research fellowship. We also thank the University Grants Commission, India, for providing a Junior Research Fellowship to VVS.
Funding
This work was financially supported by an ECRA grant from the Science and Engineering Research Board (ECR/2017/001018), the Government of India, a Start Up Grant (Dev. Scheme No. 5007), and a Research Grant for Faculty (IoE Dev. Scheme-6031) from Banaras Hindu University to PA.
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Shambhu Kumar Prasad: investigation, formal analysis, validation, visualization, writing-original draft. Vishal Vikram Singh: investigation, writing-review. Arup Acharjee: conceptualization, formal analysis, writing-review, and editing. Papia Acharjee: conceptualization, funding acquisition, formal analysis, supervision, writing-review, and editing.
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Prasad, S.K., Singh, V.V., Acharjee, A. et al. Elucidating hippocampal proteome dynamics in moderate hepatic encephalopathy rats: insights from high-resolution mass spectrometry. Exp Brain Res (2024). https://doi.org/10.1007/s00221-024-06853-4
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DOI: https://doi.org/10.1007/s00221-024-06853-4