Histomorphometric analysis
Heart tissue was fixed in 10% phosphate-buffered formalin for
1 day and processed in a routine manner to generate 5-μm-thick
paraffin sections. Sections were stained with Masson-Trichrome
staining and examined microscopically. Approximately 10 randomly
selected fields of each heart sections were observed with a light
microscope at 100 × magnification. The occurrence of myocardial
fibrosis was examined in each heart section and the percentage
estimated for every field. Myocardium thickness and lumen
diameter of the left ventricle (LV) were measured with a ruler,
and images were captured at 400 × magnification on a flat
computer monitor. Five randomly selected images of the LV from
each heart were used for measuring myocardial thickness and
the diameter of LV was measured at the largest and the smallest part.