Anat Cell Biol.  2020 Jun;53(2):169-182. 10.5115/acb.19.231.

Effect of meloxicam (cyclooygenase-2 inhibitor) versus vitamin D3 (cholecalciferol) as ameliorating agents of progressive doxorubicin-induced nephrotoxicity in rats

• Elsherbini DMA
• Ebrahim HA

• KMID: 2503447
• DOI: http://doi.org/10.5115/acb.19.231

Abstract

Doxorubicin (DOX)-induced nephropathy hampered its antineoplastic efficiency. The objective of the current work is to assess the prospective ameliorating effects of meloxicam versus vitamin D3 (Vit D3, cholecalciferol) against progressive DOX-induced nephropathy in rats trying to ascertain the possible mechanism underlying such amelioration. Ninety Male Wistar rats were randomly distributed to five experimental groups for 3 weeks, with saline, meloxicam (daily), DOX (single dose), Vit D3+DOX, or both meloxicam and DOX. We measured levels of urinary protein, serum creatinine, malondialdehyde (MDA) and renal reduced glutathione (GSH). In addition, tumor necrosis factor-alpha (TNF-α) expression and renal histopathology were assessed. Meloxicam alone treated group revealed no significant difference in urinary protein and serum creatinine. It also presented non-significant reduction in the MDA content while an increase in the reduced GSH content in contrast to the control group, which is more evident after the first week. Renal sections of rats received meloxicam only showed no significant histological changes and negative immunoreactivity compared to the control group. DOX induced a significant increase in urinary protein, serum creatinine, decrease reduced GSH, increased renal MDA and disrupted renal morphometric parameters and histology with increased TNF-α expression. Combination groups of Vit D3+DOX and meloxicam+DOX showed improvement of all DOX disturbed parameters. Meloxicam showed better results most likely due to anti-inflammatory and antioxidant activities superimposing the immune-modulatory effect of Vit D3. So, it is recommended to use meloxicam in patients receiving DOX as a renoprotective agent in addition to its analgesic effects required by cancer patients.

Keyword

Antioxidants; Meloxicam; Nephropathy; Doxorubicin

Figure

• 

  Fig. 1 Total urinary protein (A), serum creatinine (B), MDA level in renal tissue (C), and GSH in renal tissue (D) for groups with saline treated (●), meloxicam (■), DOX (▲), DOX+Vit D3 (▽) and DOX+meloxicam (□) treated groups graphed versus time. Values are presented as means±SD. DOX, doxorubicin; GSH, glutathione; MDA, malondialdehyde; Vit D3, vitamin D3. *P, significant difference between combined DOX+Vit D3 vs. combined DOX+meloxicam treated group; **P, significant in contrast to other groups.

• 

  Fig. 2 Representative of cortical kidney sections stained by H&E from rats representing groups of control, meloxicam, DOX, DOX+Vit D3, and DOX+meloxicam after 1st week. Control sections revealed the normal histological architecture, renal T and renal C (A, ×100), F and PCT (B, ×400). Meloxicam revealed no changes (C, ×100). The renal glomeruli and tubules appear to great extent normal (D, ×400). DOX is showing decrease number of renal C in contrast to other groups (E, ×100), glomerular E, synechia of the glomerular tuft with Bowman’s capsule (➨), mesangial matrix increment (➤), and dilation of the F (🟌). Some cells of PCT loss its brush borders and some nuclei appear pyknotic (→) (F, ×400). Conversely, combination group revealed rare disruption of some renal C (G, I, ×100). Less dilation of F (🟌). Most of PCT appears normal (T) lower number of cells of PCT showed loss of brush border and pyknosis (→) compared to the DOX treated group (H, J, ×400). C, corpuscles; DOX, doxorubicin; E, edema; F, filtration space; PCT, proximal convoluted tubules; T, tubules; Vit D3, vitamin D3.

• 

  Fig. 3 After 2nd week: Sections of control group demonstrating the normal histological architecture, renal T, renal C (A, ×100), and F (B, ×400). No difference is shown in meloxicam (C, ×100). The renal glomeruli and tubules appear normal to great extent (D, ×400). DOX is showing shrinkage of renal C (⮏) compared to other groups (E, ×100), some showing glomerular E and most cells of PCT loss its brush borders and some nuclei appear psychotic (→) (F, ×400). However, combination group exhibiting less disruption of some renal C (G, I, ×100). Most of PCT appears normal (T) lower number of cells of PCT showed loss of brush border and pyknosis (→) compared to the DOX treated group (H, J, ×400), with better picture in DOX+meloxicam compared with DOX+Vit D3 group. C, corpuscles; DOX, doxorubicin; E, edema; F, filtration space; T, tubules; Vit D3, vitamin D3.

• 

  Fig. 4 After 3rd week Sections of control group exhibiting the normal histological architecture, renal C, renal T (A, ×100), and F (B, ×400). Meloxicam is showing no difference from the normal (C, ×100). The renal glomeruli and tubules appear to great extent normal (D, ×400). DOX is showing prominent shrinkage of renal C (⮏) compared to other groups and vascular congestion (↯) (E×100), and dilation of the F (🟌). Most cells of PCT loss its brush borders, dilated and some nuclei appear psychotic (→) (F×400). However, combination group showing less distortion of renal C (G, I, ×100). Most prominent finding is dilation of T and loss of brush borders (→) compared to the control group (H, J, ×400), with better picture in DOX+meloxicam compared with DOX+Vit D3 group. C, corpuscle; DOX, doxorubicin; F, filtration space; PCT, proximal convoluted tubules; T, tubules.

• 

  Fig. 5 Representatives of renal cortical tissue stained by TNF-α of: (A–C), control and meloxicam groups (D–F) after 1st, 2nd, and 3rd weeks, respectively displaying negative expression. DOX treated animals displayed intensive expression (G–I) after (1st, 2nd, and 3rd weeks respectively) in the renal glomeruli (→) and renal tubules (➤). Combined DOX-Vit D3 treated rats displayed moderate expression (J–L) after (1st, 2nd, and 3rd weeks respectively) within the glomeruli (→) while mild expression in the renal tubules (➤) .DOX/meloxicam treated groups demonstrated considerable improvement with mild expression (M–O) after (1st, 2nd, and 3rd weeks respectively) in the renal glomeruli (→) and renal tubules (➤). The expression is predominantly cytoplasmic, but with few nuclear expression. Immunohistochemistry counter stained with H&E, ×400. DOX, doxorubicin; TNF-α, tumor necrosis factor-alpha; Vit D3, vitamin D3.

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