Plant Natural Products Calycosin and Gallic Acid Synergistically Attenuate Neutrophil Infiltration and Subsequent Injury in Isoproterenol-Induced Myocardial Infarction: A Possible Role for Leukotriene B4 12-Hydroxydehydrogenase?

Figure 5

In vivo cardioprotective effects of calycosin and gallic acid in combination. (a) H&E histopathological staining of mouse heart tissues. Five groups () of C57BL/6 mice were treated with vehicle and isoproterenol (ISO); ISO + GA; ISO + CA and ISO + GA + CA. The cardiac tissues were stained with H&E. Representative images were selected from five groups. (b) Immunohistofluorescence staining of neutrophil biomarker Ly6G in mouse heart tissues. Following the treatment as described in (a), the heart tissues were probed with Ly6G antibody and subsequently detected with Alexa Fluor 488-labeled secondary antibody. The cell nuclei were stained with DAPI. The images were captured under a Zeiss fluorescence microscope. Representative images were selected from five groups. (c) Quantitation of neutrophil infiltration. Ly6G-positive cells were counted under a Zeiss fluorescence microscope. The mean numbers of neutrophils were shown and analyzed by one-way ANOVA followed by Tukey’s test. (Drugs + ISO versus ISO group). (d) MPO activity in the cardiac tissues. Following the treatment as described in (a), the heart tissues were homogenized for the assay of MPO activity. The values represent the mean ± SD. (Drugs + ISO versus ISO group). (e) MDA levels in the cardiac tissues. Following the treatment as described in (a), the heart tissues were homogenized for the determination of MDA levels. The values represent the mean ± SD. ; (Drugs + ISO versus ISO group).

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