Figure 4. Effect of pirfenidone on myocardial inflammation (day 4) after I/R injury.

Wild-type mice were subjected to 90 minutes closed-chest ischemia followed by reperfusion (I/R injury). Mice were fed either chow enriched with pirfenidone (PFD) or regular chow. Mice were sacrificed at day 4 after I/R injury and the heart was collected for analysis via flow cytometry. n = 8 I/R-control, n = 4 I/R-PFD. (A) Total number of CD45⁺ cells/mg heart tissue. (B) Leukocyte subsets in the myocardium (percentage of total: CD19⁺, Ly6g⁺, Ly6C⁺CD64^lo/–, CD64⁺Ly6C^lo; n = 4/group). (C) Representative FACS analysis of MHC-II and CCR-2 macrophages and monocytes. (D) Macrophage/monocyte subsets in the myocardium as defined by expression of CCR 2, low (l) or high (h) and MHC-II expression, low (l) or high (h). Percentage of total, n = 4/group.***P < 0.001. Bars represent the mean, and error bars represent standard deviation. P values were calculated with Student’s t test.