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The Genetic Evidence of Burn-Induced Cardiac Mitochondrial Metabolism Dysfunction.

ABSTRACT: Burn-induced cardiac dysfunction is thought to involve mitochondrial dysfunction, although the mechanisms responsible are unclear. In this study, we used our established model of in vivo burn injury to understand the genetic evidence of burn-induced mitochondrial confusion dysfunction by describing cardiac mitochondrial metabolism-related gene expression after burn. Cardiac tissue was collected at 24 hours after burn injury. An O2K respirometer system was utilized to measure the cardiac mitochondrial function. Oxidative phosphorylation complex activities were determined using enzyme activity assays. RT Profiler PCR array was used to identify the differential regulation of genes involved in mitochondrial biogenesis and metabolism. The quantitative qPCR and Western blotting were applied to validate the differentially expressed genes. Burn-induced cardiac mitochondrial dysfunction was supported by the finding of decreased state 3 respiration, decreased mitochondrial electron transport chain activity in complex I, III, IV, and V, and decreased mitochondrial DNA-encoded gene expression as well as decreased levels of the corresponding proteins after burn injury. Eighty-four mitochondrial metabolism-related gene profiles were measured. The mitochondrial gene profile showed that 29 genes related to mitochondrial energy and metabolism was differentially expressed. Of these 29 genes, 16 were more than 2-fold upregulated and 13 were more than 2-fold downregulated. All genes were validated using qPCR and partial genes were correlated with their protein levels. This study provides preliminary evidence that a large percentage of mitochondrial metabolism-related genes in cardiomyocytes were significantly affected by burn injury.

SUBMITTER: Wen JJ

PROVIDER: S-EPMC7761708 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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The Genetic Evidence of Burn-Induced Cardiac Mitochondrial Metabolism Dysfunction.

Wen Jake J JJ Cummins Claire B CB Williams Taylor P TP Radhakrishnan Ravi S RS

Biomedicines 20201203 12

Burn-induced cardiac dysfunction is thought to involve mitochondrial dysfunction, although the mechanisms responsible are unclear. In this study, we used our established model of in vivo burn injury to understand the genetic evidence of burn-induced mitochondrial confusion dysfunction by describing cardiac mitochondrial metabolism-related gene expression after burn. Cardiac tissue was collected at 24 hours after burn injury. An O2K respirometer system was utilized to measure the cardiac mitochon ...[more]

PMID: 33287280

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Project description:Abstract Introduction The cytokine cascade is a prime factor in inflammation following burn injury, leading to cardiac dysfunction. Studies on immune-related key cardiac genes after burn injury remain limited. The TLRs act individually or as heterodimers, interacting with adaptor proteins to initiate MyD88 or TICAM1 (TRIF) dependent responses. This results in signaling cascades through NFkB, which activates downstream JNK/p38 signaling or cytokine secretion. Dysregulation of TLR and related signaling pathways has severe consequences and is implicated in autoimmune diseases and chronic inflammation. The objective of this study is to identify the immune-related key genes and explore the pathways by microarray specific to members of the TLR signaling family as well as adaptor and effector proteins, and downstream members of TLR activation including the NFkB, JNK/p38, IRF and JAK/STAT signaling pathways. Methods The microarray was applied to profile expression of 84 genes inTLR-mediated signal transduction and innate immunity. The Qiagen web-based tools in GeneGlobe Data Analysis Center was used to analyze burn-induced up-regulation of these genes. String-DB version 11.5 was employed to analyze differentially expressed gene function and interaction. Real-time qPCR was utilized to validate the data. GO enrichment analysis (FDR< 0.05) using "clusterProfiler" predicted the roles of immune-key genes in the 3 aspects of biological processes (BP), cellular components (CC), and molecular functions (MF). The biological pathway analysis was performed to obtain the pathways. The protein-protein interaction (PPI) network was built using "Multiple Protein" of the string database (http://stringdb.org/, version 15.0). The DEGs were visualized by using the qPCR. Results 32 up-regulated genes were identified. Functional enrichment analysis showed that these genes were enriched in cytokine receptor activity, immune receptor activity, and integrin family cell surface interactions.14 of 32 genes belong to the TLR signaling pathway; 5 of 32 genes to cytokines and inflammation; 5 of 32 genes belong to apoptosis; 6 in the MARP signaling pathway; 4 genes belong to the IL-1 signaling pathway; 4 genes in the IL-6 signaling pathway); 3 genes in toIL-2 signaling pathway and 3 genes belong to oxidative stress responses. Process study demonstrated the genes are associated with regulation of cytokine production (GO:0001817), immune response (GO:0006955), and defense response (GO:0006952). Conclusions This study provides evidence that microarray plays a role in identification of burn-induced cardiac dysfunction DEGs. Deep analysis of up-regulated genes after burn injury supported the idea that the proinflammatory cascade results in cardiac inflammation.

Project description:An elevated level of homocysteine, a thiol amino acid, is associated with various complex disorders. The cellular effects of homocysteine and its precursors S-adenosylhomocysteine (AdoHcy) and S-adenosylmethionine (AdoMet) are, however, poorly understood. We used Saccharomyces cerevisiae as a model to understand the basis of pathogenicity induced by homocysteine and its precursors. Both homocysteine and AdoHcy but not AdoMet inhibited the growth of the str4? strain (which lacks the enzyme that converts homocysteine to cystathionine-mimicking vascular cells). Addition of AdoMet abrogated the inhibitory effect of AdoHcy but not that of homocysteine indicating that an increase in the AdoMet/AdoHcy ratio is sufficient to overcome the AdoHcy-mediated growth defect but not that of homocysteine. Also, the transcriptomic profile of AdoHcy and homocysteine showed gross dissimilarity based on gene enrichment analysis. Furthermore, compared with homocysteine, AdoHcy treatment caused a higher level of oxidative stress in the cells. However, unlike a previously reported response in wild type (Kumar, A., John, L., Alam, M. M., Gupta, A., Sharma, G., Pillai, B., and Sengupta, S. (2006) Biochem. J. 396, 61-69), the str4? strain did not exhibit an endoplasmic reticulum stress response. This suggests that homocysteine induces varied response depending on the flux of homocysteine metabolism. We also observed altered expression of mitochondrial genes, defective membrane potential, and fragmentation of the mitochondrial network together with the increased expression of fission genes indicating that the imbalance in homocysteine metabolism has a major effect on mitochondrial functions. Furthermore, treatment of cells with homocysteine or AdoHcy resulted in apoptosis as revealed by annexin V staining and TUNEL assay. Cumulatively, our results suggest that elevated levels of homocysteine lead to mitochondrial dysfunction, which could potentially initiate pro-apoptotic pathways, and this could be one of the mechanisms underlying homocysteine-induced pathogenicity.

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The Genetic Evidence of Burn-Induced Cardiac Mitochondrial Metabolism Dysfunction.

Publications

The Genetic Evidence of Burn-Induced Cardiac Mitochondrial Metabolism Dysfunction.

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