Immunological characterization of stroke-heart syndrome and identification of inflammatory therapeutic targets

Figure 1. Stroke induced heart injury. (A) The cerebral infarct volume was determined by TTC staining. (B) mNSS scores of mice measured 24 h after MCAO. (C) Serum cTnI levels determined by ELISA. (D–E) Leukocyte infiltration into myocardium (D) and quantification (E) measured by IF with anti-CD45 antibody. *P<0.05.

Figure 2. Identification of DEGs in the myocardium of SHS- and sham-treated mice in the GEO GSE102558 dataset. (A) Volcano plot of gene expression levels in SHS- and sham-treated mice. (B) A total of 138 DEGs were identified, including 91 upregulated genes and 47 downregulated genes in SHS- versus sham-treated mice. (C) The box diagram of data in the SHS and sham datasets. (D) Representative heatmap of the top 20 genes most upregulated and most downregulated DEGs.

Figure 3. GO and KEGG analysis of DEGs in myocardium of MCAO- and sham-treated mice. (A, B) Significantly enriched pathways determined by GO and KEGG analysis using MetaScape. (C) The enriched ontology clusters colored by p-value. (D–G) Chordal graph of BPs (D), CCs (E), and MFs (F) from GO analysis. (G) Chordal graph of enriched pathways determined by KEGG analysis.

Figure 4. Analysis of immune cell infiltration in cardiac tissue of SHS- and sham-treated mice. (A) The fraction of 25 subsets of immune cells in SHS and sham samples. (B) The box graph shows the difference in populations of infiltrating immune cells between SHS and sham samples. (C) Correlation analysis of 25 types of immune cells. *P<0.05, **P<0.01.

Figure 5. DIREGs in MCAO versus sham mouse myocardium. (A) Venn diagram showing intersection between138 DEGs and the HALLMARK_INFLAMMATORY_RESPONSEgenes to identify DIREGs. (B) Chord diagram of correlation analysis of DIREGs.

Figure 6. Correlation between DEIRGs and immune cells infiltrating the myocardium of MCAO-treated mice. (A–E) Correlation between expression of Aplnr (A), Ccrl2 (B), Cdkn1a (C), Irak2 (D), and Serpine1 (E) and immune cell populations.

Figure 7. RT-qPCR of differentially expressing inflammation-related genes (DEIRGs) in myocardium of MCAO- and sham-treated mice. (A–E) Transcriptional expression of Aplnr (A), Ccrl2 (B), Cdkn1a (C), Irak2 (D) and Serpine1 (E) at 1, 3, 6, 12, and 24 h of perfusion or sham surgery. *P<0.05 compared with sham-treated mice.

Figure 8. Western blot analysis of DEIRG protein levels in myocardium of MCAO- and sham-treated mice. Western blot image of bands (three replicates each) and quantitation of signal intensities normalized with GAPDH in fold-change for APLNR (A), CDKN1A (B), IRAK2 (C), and SERPINE1 (D). *P<0.05.