LAUSR

Cumulative evidences have shown that IL-6 in atrium might play an important role in the pathogenesis of postoperative AF (POAF) via activation of atrial fibrosis in patients undergoing CABG. However, whether atria produces IL-6 after the stimulation of CABG and its causal relationship with spontaneous POAF (sPOAF) and its specific pathways is still unclear. To test the hypothesis that atrium will produce IL-6 after CABG and causes sustained sPOAF (ssPOAF) through activating pSTAT3-mediated fibroblast proliferation. To determine the causal relationship between IL-6 and sPOAF, IL-6−/− and wild type (WT) mice were both divided into three groups (10 mice/group): CABG group (NAI, mimic CABG), anti-inflammatory group (AI, mimic CABG with pericardial administration of methylprednisolone for 3 days via chest tube), and control group (anesthesia only). Mice were monitored for ssPOAF for 7 days using implanted telemetry device. Another two sets of mice, using the same models mentioned above, were euthanatized at 48th hours postoperatively. The atria of one set animals were excised and separated into pericardium (PC), pulmonary vein (PV), left atrium (LA), and right atrium (RA) and cultured for 4 hours. IL-6 levels in the supernatant were measured at 10 min and 4 hours of culture using ELISA test. The region producing the largest amount of IL-6 in the other set of animals was harvested for analyzing expressions of IL-6, pSTAT3/STAT3, connexin 43 and 40, fibroblast deposition, and collagen I and III. Path analysis was performed to determine the causal relationship of CABG induced IL-6 release, pSTAT3/fibroblast signaling, and the onset of ssPOAF. 40% NAI-WT mice developed ssPOAF (Figure 1A) which was completely protected in IL-6−/− and AI groups. IL-6 was produced by all 4 atrial regions at 4hrs after CABG stimulation with the LA producing the highest amount. Western blotting (Figure 1B), RT-CPR, Masson staining, and immunofluorescence all showed a significantly upregulation of IL-6, pSTAT3/STAT3, fibroblasts, collagen I and III, and downregulation of Cx40 an 43 in NAI-WT mice, but not in IL-6−/− and AI mice. IL-6 was colocalized with vimentin to a large extent in cytoplasm (Figure 1C). IL-6 had strong positive correlation with pSTAT3/STAT3, collagen I and III (all r>0.700, P<0.001), moderate and weak negative correlation with Cx40 and 43 (r=−0.505, P<0.001; r=−0.307, P=0.048, respectively). Path analysis (Figure 1D) revealed that every 1 unit increase in IL-6 upregulated a 0.589 unit increase in ssPOAF, which was mediated by pSTAT3/collagen indirectly and collagen I/ collagen III directly. Our study, for the first time, to the best of our knowledge, established a novel pathophysiological role of IL-6/pSTAT3/fibroblast signaling in the pathogenesis of ssPOAF and demonstrated that inhibition of atrial IL-6 might be a potential novel sPOAF prevention strategy. The National Natural Science Foundation (No.81170170)