European Journal of Biomedical and Pharmaceutical Sciences

ANALYSIS OF RELATIVE GENE EXPRESSION OF ANTIOXIDANT ENZYMES IN ADRENAL GLAND USING REAL TIME QUANTITATIVE PCR IN EXPERIMENTAL FLUOROSIS AND MODULATORY POTENTIAL OF CURCUMIN ADMINISTRATION

A. Shashi* and Manisha Tikka

ABSTRACT

The present study was conducted to investigate the efficatory effect of Curcumin on oxidative stress, antioxidant enzymes gene expression and pathological alterations in adrenal gland of rat in sodium fluoride (NaF) intoxication. A total of 36 young Wistar albino rats were randomly divided into six groups. Each group contained six rats. The group I was given deionized water orally for 40 days. The groups II and III were administered with 300 and 600 mg of NaF/kg b.w./day for same period. The group IV was given 200 mg/kg b.w./day of Curcumin only for 20 days. The groups V and VI exposed to NaF toxicity of 300 and 600 mg/kg b.w./day for 40 days respectively were post-treated with 200 mg/kg b.w./day of Curcumin for 20 days. At the end of experimental period, animals were sacrificed under anaesthesia and adrenal gland was removed and processed for biochemical, molecular and histopathological analysis. The levels of malondialdehyde (MDA), reduced glutathione (GSH) and the activities of antioxidant enzymes such as mitochondrial superoxide dismutase 2 (MnSOD or SOD 2), glutathione peroxide (GPx), and catalase (CAT) were determined. The gene expression of mitochondrial SOD 2 and GPx was evaluated by using real time RT-PCR. The histopathological alterations in adrenal gland tissue were examined using hematoxylin and eosin staining technique. Results revealed that NaF treatment induced significant (P<0.0001) elevation in lipid peroxidation and SOD 2 activity while significant (P<0.0001) reduction in the activities of GPx, CAT and GSH content was observed. The relative gene expression of SOD 2 was significantly (P<0.05-0.01) upregulated, where as GPx was significantly (P<0.05-0.01) downregulated on fluoride exposure. Histopathological evaluation also revealed that NaF intoxication resulted into loss of cellular architecture and distorted cell behaviours including ruptured cell membrane, pyknotic nuclei, and necrosis in the adrenal gland cells. Interestingly, Curcumin supplementation markedly minimized the biochemical and molecular changes and also normalized the cellular adversities caused by the NaF exposure in adrenal gland of rat and, therefore, showed ameliorative effect by eliminating the ill effects.

Keywords: Adrenal gland; Antioxidant enzymes; Curcumin; Gene expression; Oxidative stress, Fluorosis.