Pathological Investigation of the Effect of Bovine Colostrum Against 5-FU-Induced Liver, Kidney, and Heart Toxicity in Rats

Abstract

This study aimed to investigate the possible histopathological and immunohistochemical effects of bovine colostrum (BC) against the toxic effects of 5-fluorouracil (5-FU) on the liver, kidney, and heart of Wistar Albino rats. Animals were divided into three groups: control, 5-FU, and 5-FU+BC. The control group received 2 mL/kg i.p. saline, the 5-FU group 100 mg/kg i.p. 5-FU, and the 5-FU+BC group received 100 mg/kg i.p. saline on the first day of the study. The 5-FU and 5-FU+BC groups received 100 mg/kg i.p. of 5-FU and 1000 mg/kg BC orally each day of the study. Liver, kidney, and heart tissues were examined histopathologically for lesions and the expression of TNF-α, HSP-27, CASP-3, and 8-OHdG. No pathologic lesions were observed in the control group, whereas severe pathologic lesions were observed in the 5-FU group. In the 5-FU+BC group, the lesions were less severe than in the 5-FU group. In immunohistochemical examination, biomarker expression was not observed in the control group, whereas it was severe in the 5-FU group and less severe in the 5-FU+BC group. At the end of the study, it was observed that 5-FU-induced pathological findings in liver, kidney, and heart tissues decreased with the use of bovine colostrum. The difference between the control group and the 5-FU and 5-FU+BC groups was significant (p < 0.01 and p < 0.05, respectively). Although the BC addition did not show any statistical significance in the pathological scores of 5-FU in liver, kidney, and heart tissues, it was observed that it improved the lesions of these tissues. Nevertheless, histopathological and immunohistochemical analyses showed visible improvements in the 5-FU+BC group. Although more studies are needed, it is hoped that BC will improve prognosis by both reducing the side effects of 5-FU, a good chemotherapeutic agent, and its antineoplastic properties.

Keywords: 5-FU; cancer; cardiotoxicity; hepatotoxicity; nephrotoxicity; pathology.

Figure 1

Histopathologic sections of liver (HE, X200): (A) control group with normal histological appearance, (B) 5-FU group with degenerative-necrotic lesions in hepatocytes (open arrowhead), inflammatory cell infiltrations (arrowhead), vascular changes (arrows), and (C) 5-FU+BC group with degenerative-necrotic lesions in hepatocytes (arrowheads), inflammatory cell infiltrations (arrowhead), and vascular changes (arrow).

Figure 2

Graphical representation of liver pathology scoring for (A) DNL, (B) ICI, (C) VC, and (D–F) their relative fold change. * p < 0.05, ** p < 0.01. DNL (degenerative-necrotic lesion), ICI (inflammatory cell infiltrate), VC (vascular change).

Figure 3

Histopathologic sections of kidney (HE, X200): (A) control group with normal histological appearance, (B) 5-FU group with degenerative-necrotic lesions in tubular epithelium (open arrowheads), inflammatory cell infiltrations (arrowheads), vascular changes (arrows), and (C) 5-FU+BC group with degenerative-necrotic lesions in tubular epithelium (open arrowheads), inflammatory cell infiltrates (arrowheads), and vascular changes (arrow).

Figure 4

Graphical representation of kidney pathology scoring for (A) DNL, (B) ICI, (C) VC, and (D–F) their relative fold change. * p < 0.05, ** p < 0.01.

Figure 5

Histopathologic sections of heart (HE, X200): (A) control group with normal histological appearance; (B) 5-FU group, with degenerative-necrotic lesions in cardiomyocytes (open arrowheads), inflammatory cell infiltrates (arrowheads), vascular changes (arrows), and (C) 5-FU+BC group with degenerative-necrotic lesions in cardiomyocytes (open arrowheads), inflammatory cell infiltrates (arrowheads), and vascular changes (arrow).

Figure 6

Graphical representation of heart pathology scoring for (A) DNL, (B) ICI, (C) VC, and (D–F) their relative fold change. * p < 0.05, ** p < 0.01.

Figure 7

Immunohistochemical sections of the liver (IHC, X200): (A) control group TNF-α immunonegative; (B) 5-FU group with severe TNF-α expression, (C) 5-FU+BC group with TNF-α expression, (D) control group HSP-27 immunonegative (E) 5-FU group with severe HSP-27 expression, (F) 5-FU+BC group with HSP27 expression, (G) control group CASP-3 immunonegative, (H) 5-FU group with severe CASP-3 expression, (I) 5-FU+BC group with CASP-3 expression, (J) control group 8-OHdG immunonegative, (K) 5-FU group with severe 8-OHdG expression, and (L) 5-FU+BC group with 8-OHdG expression.

Figure 8

Graphical representation of (A) TNF-α, (B) HSP-27, (C) CASP-3, (D) 8-OHdG liver pathology scoring, and (E–H) their relative fold change. * p < 0.05, ** p < 0.01.

Figure 9

Immunohistochemical sections of the kidney (IHC, X200): (A) control group TNF-α immunonegative; (B) 5-FU group with severe TNF-α expression, (C) 5-FU + BC group with TNF-α expression, (D) control group HSP-27 immunonegative, (E) 5-FU group with severe HSP-27 expression, (F) 5-FU+BC group with HSP27 expression, (G) control group CASP-3 immunonegative, (H) 5-FU group with severe CASP-3 expression, (I) 5-FU+BC group with CASP-3 expression, (J) control group 8-OHdG immunonegative, (K) 5-FU group with severe 8-OHdG expression, and (L) 5-FU+BC group with 8-OHdG expression.

Figure 10

Graphical representation of (A) TNF-α, (B) HSP-27, (C) CASP-3, (D) 8-OHdG kidney pathology scoring, and (E–H) their relative score. * p < 0.05, ** p < 0.01.

Figure 11

Immunohistochemical sections of the heart (IHC, X200): (A) control group TNF-α immunonegative; (B) 5-FU group with severe TNF-α expression, (C) 5-FU+BC group with TNF-α expression, (D) control group HSP-27 immunonegative, (E) 5-FU group with severe HSP-27 expression, (F) 5-FU+BC group with HSP27 expression, (G) control group CASP-3 immunonegative, (H) 5-FU group with severe CASP-3 expression, (I) 5-FU+BC group with CASP-3 expression, (J) control group 8-OHdG immunonegative, (K) 5-FU group with severe 8-OHdG expression, and (L) 5-FU+BC group with 8-OHdG expression.

Figure 12

Graphical representation of (A) TNF-α, (B) HSP-27, (C) CASP-3, (D) 8-OHdG heart pathology scoring, and (E–H) their relative fold change. * p < 0.05, ** p < 0.01.