Study of the protective effects of cyanocobalamin on methotrexate induced nephrotoxicity in rats

Abstract

Background: Methotrexate (MTX) is a chemotherapeutic drug, used mainly in many cancerous stages, inflammatory and auto-immune diseases, but its use has been limited by its nephrotoxicity. Cyanocobalamin is a water-soluble vitamin possessing nephro-protective properties. The aim of this study was to investigate the effect of cyanocobalamin on the nephrotoxicity of methotrexate. Methods: In the study 42 albino adult female rats were used, divided into six groups each containing seven rats (n=7). 1st group: Control group (Negative control), 7 rats were injected intraperitoneally with 0.5ml/kg/day NS. Second group: 7 rats were injected intraperitoneally with a single dose of methotrexate (20 mg/kg) for 4 days. Third Group: 7 rats were given intraperitoneally cyanocobalamin at a dose (1.5 mg/kg/day) for two weeks, fourth, fifth, sixth group: 7 rats from each group were injected intraperitoneally with different concentrations of cyanocobalamin (0.5, 1, 1.5 mg/kg /day), respectively, for two weeks and MTX (20 mg/kg), which was injected only on day 11. On day 15, rats from all groups were euthanized, and blood samples were taken for biochemical tests, including evaluating serum urea and creatinine. The kidneys were extracted for histological investigation and evaluation of antioxidant (GSH) and oxidative stress (MDA) by using kidney tissue homogenates. Results: This study revealed that kidney damage, produced by the MTX (group II), is manifested by significantly elevated (P<0.05) urea and creatinine. On the contrary, the cyanocobalamin groups (IV, V, VI) significantly (P<0.05) reduced urea and creatinine. Renal antioxidant defense systems, such as reduced glutathione depleted by MTX therapy, were restored to normal levels by cyanocobalamin. Furthermore, cyanocobalamin reduced oxidative stress (MDA) and histologically reduced renal tissue injury induced by MTX. Conclusions: In conclusion, the study revealed that cyanocobalamin has a nephroprotective action upon MTX-induced renal damage in rats; cyanocobalamin may offer a protective effect, such as antioxidant action.

Study of the protective effects of cyanocobalamin on methotrexate induced nephrotoxicity in rats

“…Effect of cyanocobalamin on GSH contents within kidney tissue homogenate of rats A non-significant difference was observed between the cyanocobalamin (group III) and control groups (group I). 37 While the level of GSH in the renal tissue homogenate significantly increased (P<0.05) in both groups IV and V, it was highly significantly increased in group VI compared to the group treated with MTX treated group (P<0.01) as shown in Figure 1.…”

“…Group IV showed a moderate increase in urinary space, moderate congestion of the glomeruli and interstitial as shown in Figure 8A, a moderate amount of hyaline cast formation and degeneration as shown in Figure 8B, moderate inflammation of the pelvicalyceal system Figure 8c, moderate infiltrate of inflammatory cells of the interstitial Figure 8d as shown in the Extended data. 39 Group V showed a moderate increase in urinary space, moderate congestion of glomeruli and interstitial as shown in Figure 9A, moderate amount of hyaline cast in the tubular lumen and degeneration as shown in Figure 9B, moderate inflammation of the pelvicalyceal system 9C, moderate inflammatory cell infiltrate of interstitial as shown in Figure 9D as shown in the Extended data. 39 Furthermore, in group VI there was a significant decrease in damage in all histopathological parameters compared to other groups (II, IV, and V), There is mild glomerular congestion and increased cellularity, the interstitial space also shows mild congestion and degeneration, mild increased urinary space as shown in Figure 10A, Few casts are seen within tubular lumens as shown in Figure 10B, mild inflammatory infiltrate in the pelvicalyceal system as shown in Figure 10c, mild inflammatory cell infiltrate of the interstitial as shown in Figure 10d in the Extended data.…”

“…18 At the end of this experiment and on day 15 the rats were euthanized by the use of xylazine (10 mg/kg) and ketamine (75 mg/kg) 19 both are commonly used for this purpose (this is followed by the ethical standards and ARRIVE guidelines, and this type of euthanization is normally performed in most acute toxicity tests). Kidney tissue samples were taken for biochemical 37 and histopathological studies. 38 Sample collection and tissue preparation After euthanasian of the animals, blood was collected by intracardiac puncture.…”

“…Kidney tissue samples were taken for biochemical 37 and histopathological studies. 38 Sample collection and tissue preparation After euthanasian of the animals, blood was collected by intracardiac puncture. The clot was isolated with a glass rod and then centrifuged (CGOLDENWALL 80-2 Electric Lab Centrifuge) at 3000 rpm for 15 minutes, the supernatant was used to evaluate urea and creatinine as parameters of kidney function.…”

“…https://doi.org/10.5281/zenodo.6982116. 40 Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).…”