Protective effect of diosmin against doxorubicin-induced nephrotoxicity

Abstract

Doxorubicin (Dox) is an anthracycline antibiotic that is primarily used for treating various solid tumors including that of pulmonary, ovary, breast, uterine, cervix, and several blood cancers. However, nephrotoxicity associated with Dox treatment limits its clinical use. Administration of Dox in combination with compounds exhibiting antioxidant properties are being used to minimize the side effects of Dox. Diosmin is a flavonoid glycoside with numerous beneficial properties that is found in the pericarp of many citrus fruits. Diosmin has demonstrated antioxidant, anti-inflammatory, and anti-apoptotic effects in response to various insults, although the exact mechanism remains unknown. Therefore, this study was designed to evaluate the effect of diosmin in preventing kidney damage in response to Dox treatment. Male Wistar rats were randomly divided into four groups: control group, Dox group (20 mg/kg, i.p.), Dox plus low-dose diosmin group (100 mg/kg orally), and Dox plus high-dose diosmin group (200 mg/kg orally). A single intraperitoneal injection of Dox resulted in kidney damage as evidenced by significant alterations in kidney markers, histological abnormalities, and the attenuation of antioxidant defense mechanisms (GSH, SOD, and CAT). Moreover, Dox treatment significantly altered the expression of oxidative stress, inflammatory, and anti-apoptotic protein markers. Diosmin pretreatment alleviated Dox-induced nephrotoxicity by ameliorating the antioxidant mechanism, decreasing inflammation and apoptosis, and restoring kidney architecture. In conclusion, our results indicate that diosmin is a promising therapeutic agent for the prevention of nephrotoxicity associated with DOX.

Keywords: Apoptosis; Diosmin; Doxorubicin; Inflammation; Nephrotoxicity; Oxidative stress.

Fig. 1

Structure of diosmin.

Fig. 2

Illustration of the experimental design.

Fig. 3

Pretreatment with diosmin mitigated the altered serum levels of BUN (A), creatinine (B), and albumin (C) resulting from Dox treatment. Data are presented as the mean ± SD (n = 5) where ^***p < 0.001, ^**p < 0.0, *p < 0.05, and ^NSp> 0.05. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 4

Pretreatment of diosmin diminishes the oxidative stress induced by Dox. Biochemical analysis of MDA (A), GSH (B), and CAT (C). Data are presented as the mean ± SD (n = 5) where ^**p < 0.0, *p < 0.05, and ^NSp > 0.05. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 5

Diosmin reduces oxidative stress and inflammatory protein expression and induces antioxidant proteins. (A & B) Immunoblot representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. (C–I) Graphical representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. Results are shown as the mean ± SD (n = 5) where ^***P < 0.001, ^**P < 0.01, *P < 0.05, and ^NSP > 0.05. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 5

Diosmin reduces oxidative stress and inflammatory protein expression and induces antioxidant proteins. (A & B) Immunoblot representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. (C–I) Graphical representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. Results are shown as the mean ± SD (n = 5) where ^***P < 0.001, ^**P < 0.01, *P < 0.05, and ^NSP > 0.05. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 5

Diosmin reduces oxidative stress and inflammatory protein expression and induces antioxidant proteins. (A & B) Immunoblot representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. (C–I) Graphical representation of SOD, IL-10, IL-6, NfkB-p65, TNF-α, Nox-4, and iNOS. Results are shown as the mean ± SD (n = 5) where ^***P < 0.001, ^**P < 0.01, *P < 0.05, and ^NSP > 0.05. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 6

Effect of diosmin on pro-apoptotic and anti-apoptotic protein expression. Immunoblot representation of cleaved caspase-3, Bax, and Bcl-2 (A–D). Results are shown as the mean ± SD (n = 5) where ^***P < 0.001 and ^**P < 0.01. [CON, control; DOX, doxorubicin; DOX + DS100, doxorubicin plus diosmin 100 mg/kg; and DOX + DS200, doxorubicin plus diosmin 200 mg/kg, n = number of animals].

Fig. 7

Light micrographs of kidney tissues using H&E staining. (A) The standard architecture of the kidney. (B) Disruption of the regular kidney architecture caused by Dox administration was observed as indicated by arrows. (C & D) diosmin treatment exhibited beneficial improvement in the glomeruli and tubules and tubular epithelial cell morphology at both doses.

Fig. 8

Schematic representation of doxorubicin-induced renal toxicity and underlying nephroprotective mechanism of diosmin.