Single-Cell RNA and Transcriptome Sequencing to Analyze the Role of Lactate Metabolism in Traumatic Brain Injury Astrocytes

利用单细胞RNA和转录组测序分析乳酸代谢在创伤性脑损伤星形胶质细胞中的作用

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Abstract

PURPOSE: After traumatic brain injury (TBI), ischemia and hypoxia of brain tissue, glucose undergoes anaerobic fermentation, leading to a large accumulation of lactic acid. Our aim was to explore the role of lactate metabolism in brain cells after TBI. METHOD: In scRNA-seq dataset, 10-week-old male C57BL/6 J mice were randomized to undergo mild fluid percussion injury or sham surgery, and we analyzed frontal cortex tissue during the acute (24 h) and subacute (7 days) phases of TBI at single-cell resolution. Cell cycle phases were evaluated, and principal component analysis was performed. Cell populations were identified and visualized using the UMAP downscaling technique. Differentially expressed genes (DEGs) were analyzed using the "FindAllMarkers" algorithm. In addition, the set of genes related to lactate metabolism was evaluated using the AUCell score. GO and KEGG enrichment analyses were performed to investigate the functional pathways of DEGs in astrocytes in the acute and subacute phases of TBI. RESULTS: A total of 13 cell populations were distinguished, including neurons, astrocytes, and oligodendrocyte progenitors. The number of neurons, astrocytes, and endothelial cells was reduced in the TBI group compared with the sham group. During the acute phase of TBI, enhanced interactions between brain-associated cells, especially astrocytes and oligodendrocyte precursor cells, were observed. Several signaling pathways, including EGF, CSF, MIF inflammatory factors as well as PSAP and PTN neurotrophic factor signaling were significantly enhanced after TBI. Lactate metabolism scores were elevated in the TBI group, especially in astrocytes. During the subacute phase, the frequency of intercellular communication increased but its intensity decreased. Astrocytes and oligodendrocyte precursor cells remained at high levels during both phases. PSAP signaling was closely associated with the subacute phase of TBI. Subsequently, NADH:ubiquinone oxidoreductase subunit B9 (Ndufb9) and cytochrome c oxidase subunit 8A (Cox8a) were identified as key players in lactate metabolism associated with TBI. Ndufb9 and Cox8a showed a consistent upward trend in brain tissue following TBI with transcriptomic data. CONCLUSION: Lactate metabolism genes play an important role in TBI. These findings provide new insights into the cellular and molecular mechanisms following TBI.

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