CYTOREMEDIATION OF SODIUM ARSENTIE-INDUCED TOXICITIES BY EXTRACT OF adansonia digitata LINN. IN CELL LINES AND WISTAR RATS

Abstract

Arsenicals are proven human and animal environmental clastogens and carcinogens. Research has focused on ameliorating their toxicities using medicinal plants such as Adansonia digitata (AD). Although, there are available literatures on the medicinal uses of AD, there is a dearth of information on its use in ameliorating arsenicals-induced toxicities. This study was therefore designed to investigate the cytoremediatory effects of the extracts of AD in cell lines and Wistar rats. Fruit pulp and stem bark of AD were collected from Ajibode, University of Ibadan (UI), identified at Botany Department and authenticated at Forestry Research Institute, Ibadan (FHI NO.109859). Aqueous and methanol extracts of the plant were obtained by cold maceration. The aqueous extract of the fruit pulp of AD (AEFAD) was partitioned with n-hexane, chloroform, ethyl acetate and n-butanol. Phytochemical screening of the methanol extract of the stem bark of AD (MESBAD) was determined and AEFAD was subjected to GC-MS analysis. The nitrite, 1-1-diphenyl-1-picryl-hydroxyl (DPPH), 2-2-azinibis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) scavenging and reducing power of fractions from AEFAD were assessed spectrophotometrically. Cytotoxicities and antiproliferative potential of fractions of AEFAD and MESBAD were determined in human cancer cell lines: Lung carcinoma (A-549), Breast (MCF-7), Oral (KB), Bladder (T-24), Renal (A-498) using spectrophotometry and crystal violet staining, respectively. Protein expressions of apoptosis (p21 and p53) were determined by western blotting. For the first experiment in the in vivo study, rats (100-150g) (n=45, 9 groups of equal rats) were treated thus: distilled water, Sodium arsenite (SA), Cyclophosphamide, AEFAD (200mg/kg), AEFAD 400mg/kg, SA+AEFAD (200mg/kg), SA+AEFAD (400mg/kg), Cyclophosphamide+AEFAD (200mg/kg), Cyclophosphamide+AEFAD (400mg/kg). For second experiment, rats (n=30) were treated thus: distilled water, MESBAD (400mg/kg), SA+MESBAD (400mg/kg), SA+MESBAD (300mg/kg), MESBAD (300mg/kg) and SA only). In both experiments, rats were treated orally for 14-days and sacrificed; samples of blood, bone marrow and liver collected. Alanine and aspartate aminotransferases, γ-glutamyltransferase and lipid peroxidation (LPO) were determined spectrophotometrically. Frequency of micronucleated polychromatic erythrocytes (mPCEs) and liver histology were determined microscopically. Data were analysed using descriptive statistics and ANOVA at α 0.05. Alkaloid was abundant in MESBAD (330.0±0.0mg/100g), saponins (153.3±2.9mg/100g), flavonoids (121.7±2.9mg/100g) and total polyphenols (121.7±2.9GAE/100g) were significantly high. Seven compounds were identified in AEFAD, namely: Pentadecanoic-acid, Hexadecanoic-Acid, 11-Octadecenoic-acid, Octadecanoic-acid, Oleic-acid, Nonadecanoic-acid and 3,11-Tetradecadiene-1-ol. N-butanol fraction had the highest reducing power (1.6±0.3) and ABTS scavenging capacities (78.0±2.4%). There was no significant difference between the cytotoxic effects of the fractions of AEFAD and the negative controls on the cell lines. Aqueous, ethyl acetate and n-butanol fractions conferred apoptotic morphological effects on KB. The MESBAD showed a concentration-dependent cytotoxic and antiproliferative effects on MCF-7 and, also up-regulated apoptotic markers. The AEFAD and MESBAD had hepatoprotective activities by gradual restoration of liver lesions to normal hepatocytes. Cyclophosphamide and SA increased LPO (47.8 and 36.3%, respectively), while co-treatment [(Cychlophosphamide+AEFAD) and (SA+AEFAD)] resulted in 3.1 and 1.7 folds reduction, respectively. Cyclophosphamide and SA increased mPCEs, co-treatment [(Cychlophosphamide+AEFAD), (SA+AEFAD) and (SA+MESBAD)] resulted in 2.5, 5.1 and 2.6-folds decrease, respectively.

Aqueous and methanol extracts of Adansonia digitata elicit cytoremediation in sodium arsenite-induced toxicities, while methanol extract possesses anticancer activities.