Figure 5.

Ghrelin treatment improved inflammation, oxidative stress but not apoptosis in rat lungs following CPB. (A) H&E stained lung section of a sham rat showing normal features of alveolar capillary membrane; a CPB saline group rat showing inflammatory cell infiltration (black arrows) and thickening of alveolar capillary membrane (green arrows); a ghrelin treated rat showing reduced inflammatory cell infiltration (black arrows) but thickening of the alveolar capillary membrane (green arrows). (B) Immunofluorescence staining was used to access the levels of CD-68 macrophages (red stain; white arrow) positively-stained in alveolar capillary membrane. (C) Quantitative measurement of CD-68 positive cell per 20 fields. (D) In situ TUNEL-assay was used to access the apoptotic cells per 20 fields (green stain; white arrow) in the lungs. Nuclei were labeled with 4', 6-diamidino-2-phenylindole (DAPI, blue). The merged image is presented in this figure. In all images scale bar is 50 μm. (E) Quantitative measurement of apoptotic cell number/field across 20 fields. (F) Protein expression levels of CD-68, NT and caspase-3 detected by Western blotting. β-actin was used as an internal control. (G–I) Bar graph showing the densitometric analysis of the CD-68, NT and caspase-3 Western blots. The data shown are the mean ± SEM; N = 3 rats per group. Data were analyzed by a one-way ANOVA followed by Tukey's post hoc test. *p < 0.05, **p < 0.01 vs. sham group; ^#p < 0.05 vs. CPB saline group.