At the end of measurements, the ventricular tissues were quickly removed and fixed in 4% paraformaldehyde, embedded in paraffin and sectioned at 5 microns. To identify the pro-inflammatory M1 macrophages, the ventricular samples were incubated with primary antibodies against CD68 (1:100, Abcam, Inc., UK) and inducible nitric oxide synthase (iNOS; 1:50, Abcam, Inc., UK), and cells were stained with DAPI for nuclei. The results were analyzed by Image-Pro Plus 6.0 software. More than 3 sections of ventricle from different samples were randomly selected to quantify the number of macrophages.

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