Putative Prevention of XML Injection Against Myocardial Ischemia Is Mediated by PKC and PLA2 Proteins

FIGURE 1. Protective effect of XML on isoproterenol-induced myocardial ischemia in a rat model. (A) Timeline of drug administration. CON rats (n = 5) were treated with normal saline, while test groups received an XML injection (low dose—L (n = 5), medium dose—M (n = 6), high dose—H (n = 5)), or propranolol (10 mg/kg) (n = 5) for a total of 4 days 30 min after the final administration, rats were subcutaneously injected with isoproterenol (5 mg/kg) to induce the myocardial ischemia model. Heart contractile function was assessed using the noninvasive cardiac imaging technology of echocardiography (ECG) and by measuring the levels of the cardiac troponin T (cTnT) biomarker. (B) Electrocardiogram recordings of SD rats. (C) T interval of rats in each group. (D) cTnT levels for each group. Asterisk (*) refers to statistical significance in comparisons with the CON group (*p < 0.05, **p < 0.01), while # refers to comparisons with the ISO group (# p < 0.05, ## p < 0.01,### p < 0.005).

FIGURE 2. Protective effect of XML on myocardial ischemia induced by ligation of the left anterior descending coronary artery in the rat. (A) Timeline of drug administration. Rats received a pretreatment with normal saline (n = 4) or XML injection (low dose—L (n = 5), medium dose—M (n = 6), high dose—H (n = 5)), or propranolol (10 mg/kg) (n = 6) for a total of 4 days. The left anterior descending coronary artery of rats was ligated 30 min after the final administration to induce the myocardial ischemia model. Heart contractile function was assessed by the noninvasive cardiac imaging technology of echocardiography (ECG) and measurement of serum cardiac troponin T (cTnT) biomarker levels. (B) Electrocardiogram recordings of SD rats. (C) T interval of the rats in each group. (D) cTnT levels for each group. Asterisk (*) refers to statistical significance in comparisons with the CON group (*p < 0.05, **p < 0.01, ***p < 0.005), while # refers to comparisons with the MI group (# p < 0.05, ## p < 0.01,### p < 0.005).

FIGURE 3. Overlap of genes associated with XML treatment and myocardial ischemia and their functions. (A) An intersection generated between genes related to XML-active targets and myocardial ischemia. (B) Protein–protein interaction network of the overlapped genes for XML and myocardial ischemia. (C) The top 2 MCODEs from the PPI network. (D) Most enriched KEGG pathways for XML and myocardial ischemia overlapped genes.

FIGURE 4. XML ingredient-target-pathway network. Blue circles represent active components, yellow rectangles represent target genes, and green diamonds represent pathways. The size of a node represents its degree. The larger the area, the more important the node.

FIGURE 5. Mechanisms of the protective effects of XML injection on myocardial ischemia. (A) Relative PRKCA and PRKCE transcript levels in the Sham (n = 5), MI (n = 4), and MX-M (n = 6) groups. (B,C) Relative MAPK3 (B) and PLA2G4A (C) transcript levels in the Sham (n = 5), MI (n = 4), and MX-M (n = 6) groups. (D) CYBA and CYBB gene transcript abundance in the Sham (n = 5), MI (n = 4), and MX-M (n = 6) groups. (E) Abundance of Bax and Bcl-2 proteins in the Sham, MI, and MX-M groups (n = 3). (F) Schematic model for the molecular mechanisms associated with the protective effect of XML injection against myocardial ischemia. Asterisk (*) refers to statistical significance in comparisons with the CON group (*p < 0.05, **p < 0.01, ***p < 0.005), while # refers to comparisons with the EPI group (# p < 0.05, ## p < 0.01,### p < 0.005).