The potential role of Azadirachta indica treatment on cisplatin-induced hepatotoxicity and oxidative stress in female rats

Abstract

Azadirachta indica A. Juss. (neem, family: Meliaceae) is perhaps the most commonly used traditional medicinal plant of India. In this study we investigated the protective effect of methanolic neem leaves extract (MNLE; 500 mg/Kg bwt) on rats treated with cisplatin (CDDP)-induced hepatotoxicity. Adult rats were randomly divided into four groups. CDDP was given to rats by intraperitoneal injection, while MNLE was given by oral gavage for 5 days after the CDDP injection. The injury and oxidative stress caused by CDDP on the liver and the effect of MNLE were evaluated by measuring (a) histological changes, (b) tissue biochemical oxidant and antioxidant parameters, and (c) investigating apoptosis markers immunohistochemically and by real time PCR. After treatment with MNLE, the histological damage and apoptosis induction caused by cisplatin were improved. Malondialdehyde and nitric oxide were significantly decreased; the antioxidant system, namely, glutathione content, glutathione-S-transferase, glutathione peroxidase, catalase, and superoxide dismutase activities were significantly elevated. In conclusion, MNLE may have a potential role when combined with cisplatin in chemotherapy to alleviate cisplatin-induced damage and oxidative stress in liver.

Figure 1

Histological changes in the liver of rats. (a) A control liver with normal architecture. (b) Rats treated with cisplatin with prominent inflammation and hepatocytic vacuolation. (c) Rats treated with the neem leaves extract for 5 days. (d) Rats treated with the cisplatin and neem leaves extract. Sections were stained with hematoxylin and eosin (400x).

Figure 2

Protective effects of neem leaves extract on cisplatin-induced elevation in lipid peroxidation, nitric oxide levels, and reduction in glutathione level in liver of rats. Values are means ± SEM (n = 6). ^a P < 0.05, significant change with respect to Control; ^b P < 0.05, significant change with respect to CDDP group.

Figure 3

Protective effects of neem leaves extract on cisplatin-induced reduction in superoxide dismutase and catalase activities in liver of rats. Values are means ± SEM (n = 6). ^a P < 0.05, significant change with respect to Control; ^b P < 0.05, significant change with respect to CDDP group.

Figure 4

NF-κB expression changes in the liver of rats. (a) Normal liver showing very weak expression for NF-κB. (b) Treated liver with cisplatin showing positive expression for NF-κB. (c) Neem treated liver showing negative expression for NF-κB. (d) Treated liver with cisplatin and MNLE showing medium expression for NF-κB (400x).

Figure 5

Relative quantification using RT-qPCR of mRNA expression of caspase-3, 9, and Bax genes in liver of rats treated with cisplatin and neem leaves extract. ^a P < 0.05, significant change with respect to Control; ^b P < 0.05, significant change with respect to CDDP group.