Objective: The study is to evaluate the possible genotoxic and antigenotoxic potential of Lagenandra toxicaria rhizome methanol extract using Allium cepa root tip assay. Methods: The rhizome methanol extract was prepared using Soxhlet apparatus. The A. cepa roots were treated with various concentrations of the extract at different time points and stained with aceto-orcein. The mitotic index (MI) was calculated. Results: A significant decrease in MI and increase in the percentage of clastogenicity was observed in a time- and dose-dependent manner in the roots treated with the extract at 0.2 mg/ml, 0.5 mg/ml, 1 mg/ml, 5 mg/ml, and 10 mg/ml concentration for 1, 2, and 4 h. The field emission scanning electron microscopy and Fourier-transform infrared spectroscopy revealed evident morphological and biochemical changes at 10 mg/ml treatment when compared to control for 4 h. The agarose gel electrophoresis showed loss of DNA integrity at 10 mg/ml extract for 4 h. In situ histochemical staining by Schiff’s reagent and nitroblue tetrazolium confirmed the increased lipid peroxidation and free radical generation at 4 h treatment. Subsequently, the possible antigenotoxic potential of the plant extract was explored using H2O2 standard assays. The increased percentage of H2O2 induced nuclear lesions was reduced significantly after the modulatory treatment with extract. Conclusion: The L. toxicaria rhizome methanol extract acts as an antigenotoxic agent at lower doses and at higher doses the extract induces clastogenic effects. Further studies are needed to unravel the active component in the extract that mediates the observed phenomenon in the current study.
Lagenandra toxicaria (LT) and Ariopsis peltata (AP) belongs to the family Araceae. LT is being traditionally used to cure bilious symptoms and wound healing. AP leaves are edible and the rhizome has medicinal value. With such a wide range of medicinal applications, it's essential to scientifically authenticate traditional usage and find the plant's bioactive components. Here, we have examined the anti-inflammatory and hepatoprotective potency of these plant extracts. The in-vivo liver fibrosis was induced in Wistar male rats using CCl4 and was treated with various concentrations of the methanol and water extracts of the plant rhizomes. In the in-vitro anti-inflammatory assay, LT methanol extract showed 42 % protection at 600 mg/ml which is higher than the other extracts. Among the AP and LT extracts, the 600 mg/kg methanol extract of LT treated rats showed a decreased (p<0.05) serum alanine/aspartate aminotransferase and total bilirubin levels. The plant extract brought about an amelioration of CCl4 induced pathological effects and a significant reduction in the severity of inflammation, fatty metamorphosis, necrosis and liver fibrosis. Overall, the results suggest that LT rhizome could be effective in the treatment of liver fibrosis. It is possible that this is related to the presence of anti-inflammatory and antioxidant chemicals such as 9-octadecenamide, flavonoids and phenols in the extract.
This study is the first ever approach to evaluate the possible genotoxic effect of the Lagenandra toxicaria rhizome methanol extract and its antigenotoxic potency against 3% H2O2 induced genetic damage on Allium cepa root tip model. The assay revealed a significant decrease in mitotic index (MI) and an increase in the percentage of clastogenicity in a time and dose-dependent manner in the roots exposed to Lagenandra toxicaria extract at 0.2 mg/ml, 0.5 mg/ml, 1 mg/ml, 5 mg/ml and 10 mg/ml concentration for 1, 2 and 4hour. The ultra structures of cell surface and biochemical changes of the cells were assessed in four hour treated roots using Field emission scanning electron microscopy (FESEM) and Fourier-transform infrared spectroscopy (FTIR). The higher dose of 10 mg/ml treated roots showed an evident morphological as well as biochemical changes compared to the control. The agarose gel electrophoresis showed the loss of DNA integrity in the roots that were treated with 10 mg/ml extract for four hours, where as the control showed comparatively intact DNA bands. The in situ histochemical staining by Schiff’s reagent and nitroblueterasolium (NBT) confirmed the increased lipid peroxidation and free radical generation in four hour treated samples. Subsequently, the possible antigenotoxic potential of the plant extract was explored at its lower doses using H2O2 standard assays. The H2O2 treatment induced nuclear lesions in 93.45 ± 2.33% cells and it was seen to be reduced significantly (50.99 ±7.59 % and 37.13 ± 2.66 %) after the treatment with lower concentration of 0.01 mg/ml and 0.02 mg/ml extract respectively. This suggest that the Lagenandra toxicaria rhizome methanol extract acts as antigenotoxic agent at lower doses but at higher doses the extract induces clastogenic effects and thus acts like a janus-faced compound.
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