Dabigatran mitigates cisplatin-mediated nephrotoxicity through down regulation of thrombin pathway

Abstract

Introduction: Cisplatin (CDDP) nephrotoxicity is one of the most significant complications limiting its use in cancer therapy.

Objectives: This study investigated the pivotal role played by thrombin in CDDP-mediated nephrotoxicity. This work also aimed to clarify the possible preventive effect of Dabigatran (Dab), a direct thrombin inhibitor, on CDDP nephrotoxicity.

Methods: Animals were grouped as follow; normal control group, CDDP nephrotoxicity group, CDDP + Dab 15, and CDDP + Dab 25 groups. Four days following CDDP administration, blood and urine samples were collected to evaluate renal function. Moreover, tissue samples were collected from the kidney to determine apoptosis markers, oxidative stress and histopathological evaluation. An immunofluorescence analysis of tissue factor (TF), thrombin, protease-activated receptor-2 (PAR2), fibrin, pERK1/2 and P53 proteins expression was also performed.

Results: Thrombin, pERK, cleaved caspase-3, and oxidative stress markers were significantly elevated in CDDP-treated group. However, pretreatment of animals with either low or high doses of Dab significantly improved kidney function and decreased oxidative stress and apoptotic markers.

Conclusion: We conclude that thrombin is an important factor in the pathogenesis of CDDP kidney toxicity via activation of ERK1/2, P53 and caspase-3 pathway, which can be effectively blocked by Dab.

Keywords: BUN, Blood urea nitrogen; CDDP, Cisplatin; Cisplatin; Cr, creatinine; Crcl, Creatinine clerance; Dab, Dabigatran; Dabigatran; FXa, activated form of Factor X; GSH, Reduced Glutathion; H&E, Hematoxylin–Eosin; INR, International normalized ratio; KIM-1, kidney injury molecule-1; PAR, protease-activated receptor; PAR2; Pt, Prothrombin time; Ptt, Partial thromboplastin time; ROS, Reactive oxygen species; SOD, Superoxide dismutase; TF, Tissue factor; Thrombin; pERK1/2.

Graphical abstract

Fig. 1

Effect of oral pre-treatment of animals with Dab 15 or 25 mg/kg on (A) serum Cr, (B) serum Cyst-C, (C) BUN, (D) Cr cl, (E) urinary KIM-1 and (F) Lipocaline-2 in the urine as compared to CP nephrotoxic group where (a) Differ significantly when compared to normal animals, (b) Differ significantly when compared to normal group nephrotoxic animals and (c) Differ significantly when compared to normal group CDDP + Dab 15.

Fig. 2

Effect of oral pre-treatment of animals with Dab 15 or 25 mg/kg on the expression of coagulation proteins including protease activated receptor (PAR-2) and tissue factor (TF) in renal tissue as compared to CP nephrotoxic group where (a) Differ significantly when compared to normal group, (b) Differ significantly when compared to CP + Dab 15.

Fig. 3

Effect of oral pre-treatment of animals with Dab 15 or 25 mg/kg on the expression of coagulation proteins including (A) fibrin and (B) Thrombin in renal tissue as compared to CP nephrotoxic group where (a) Differ significantly when compared to normal rats and (b) Differ significantly when compared to nephrotoxic rats.

Fig. 4

Changes of (A) pERK1/2, (B) P35 and (C) cleaved Caspase-3 proteins expression in renal tissue after oral pre-treatment of animals with Dab 15 or 25 mg/kg as compared to CP nephrotoxic group. (a) Differ significantly when compared to normal animals and (b) Differ significantly when compared to nephrotoxic group.

Fig. 5

Rat kidney tissues of (A) control group; showed normal glomerular and tubular histological features (arrow in black color). (B) CDDP nephrotoxicity group displayed severe changes in the normal architectures and epithelium cells desquamation (arrow in black color) as well as deposits of granules in the apical parts (arrow head) and C,D) CP + Dab 15 and CP + Dab 25 displayed little changes in the structural architectures (arrow head) and mild deposits of the granules in the apical parts and glomerulonephrosis (arrow).