D-Limonene Alleviates Acute Kidney Injury Following Gentamicin Administration in Rats: Role of NF-κB Pathway, Mitochondrial Apoptosis, Oxidative Stress, and PCNA

Figure 8

The effects of D-lim on immunoexpression of apoptotic biomarkers, TNF-α, and PCNA in the kidney of rats intoxicated with GM. IHC of Bax protein (a–d): a low population of only tubular cells expressing a proapoptotic marker was observed in the rat kidney related to the control (a) and D-lim (b) groups. Bax expression in both glomeruli and tubular cells enhanced in the GM group (b). Immunolabeling of an antibody in glomeruli but only in some tubules was observed in the GM+D-lim group (d). Anti-Casp-3 antibody expression (e–h): immunopositivity focused on some glomeruli and to a lesser extent tubules in the control (e) and D-lim (f) groups. A large population of epithelial cells immunoreacted to a Casp-3 antibody in the GM group (g). Decreased expression of Casp-3 in tubular cells was detected in the GM+D-lim group (h). Immunoexpression of Bcl2 (i–l): Bcl-2 has increased and has a strong cytoplasmic location in the control (i) and D-lim (j) groups but decreased in the GM group (k). Bcl2 immunoexpression considerably enhanced in the GM+D-lim group (l) compared with the GM group (k). Immunolabeling of TNF-α (m–p): low density of epithelial cells in the control (m) and D-lim (n) groups had cytoplasmic reaction. Diffuse and intensive immunostaining in the GM group (o) and focal positive reaction in the GM+D-lim group (p) were found in these IHC photomicrographs. Anti-PCNA protein (q–t): very few numbers of nuclei of epithelial cells have weak-to-moderate reaction to an antibody specified with arrows in the control (q) and D-lim (r) groups. The arrows indicate that larger numbers of nuclei are detected with a marker in the GM group (s) and the numbers of intermediate nuclei are stained in the GM+D-lim group (t).