Centella asiatica or known as ‘pegaga’ in Malaysia, is a popular medicinal herb, which is being used as main ingredient or incorporated into various herbal products. Apart from efficacy, the chemical profile and potential toxic effect of the plant are two important aspects of concern towards ensuring product satisfaction and safety of consumers. This paper reports the qualitative and quantitative chemical analysis of the leaf ethanolic extract of C. asiatica using LCMS/MS. The acute toxiciy effect of the extract and selected marker chemical constituents were further analysed using a zebrafish model. Twenty constituents, were identified and the main chemical marker constituents of the plant viz asiaticoside, asiatic acid, and madecassic acid were further quantified. Asiaticoside was found to be present in higher concentration than the other marker constituents. Meanwhile in the acute toxicity test, the LD50 of the extract on the zebrafish model was determined to be 1250 mg/L while 100% mortality was observed at the highest test concentration of 2500 mg/L. However, acute toxicity evaluation on four marker triterpenoids of the herb, i.e asiatic acid, madecassic acid, asiaticoside and madecassoside, indicated them to be quite safe on the zebrafish model, with no mortality shown for test concentrations between 10 to 500 mg/kg BW.
An ideal model organism for neurotoxicology research should meet several characteristics, such as low cost and amenable for high throughput testing. Javanese medaka (JM) has been widely used in the ecotoxicological studies related to the marine and freshwater environment, but rarely utilized for biomedical research. Therefore, in this study, the applicability of using JM in the neurotoxicology research was assessed using biochemical comparison with an established model organism, the zebrafish. Identification of biochemical changes due to the neurotoxic effects of ethanol and endosulfan was assessed using Fourier Transform Infrared (FTIR) analysis. Treatment with ethanol affected the level of lipids, proteins, glycogens and nucleic acids in the brain of JM. Meanwhile, treatment with endosulfan showed alteration in the level of lipids and nucleic acids. For the zebrafish, exposure to ethanol affected the level of protein, fatty acid and amino acid, and exposure to endosulfan induced alteration in the fatty acids, amino acids, nucleic acids and protein in the brain of zebrafish. The sensitive response of the JM toward chemicals exposure proved that it was a valuable model for neurotoxicology research. More studies need to be conducted to further develop JM as an ideal model organism for neurotoxicology research.
Depression is a common mental disorder that can adversely affect psychosocial function and quality of life. However, the exact aetiology and pathogenesis of depression are still unclear. Stress plays a major role in the pathogenesis of depression. The use of currently prescribed antidepressants has many side effects. Centella asiatica (C. asiatica) has shown promising antidepressant activity in rodent models. Here, we developed a reserpine-induced zebrafish stress-like model and performed behavioural analysis, cortisol measurement and 1H-Nuclear Magnetic Resonance (1H NMR) spectroscopy-based metabolomics analysis to test the anti-stress activity of ethanolic extract of C. asiatica (RECA). A significant increase in total distance travelled (F(8,8) = 8.905, p = 0.0054) and a reduction in freezing duration (F(9, 9) = 10.38, p = 0.0018) were found in the open field test (OFT). Asiaticoside, one of tested C.asiatica's triterpenoid gives a significant increase in contact duration (F(5,5) = 142.3, (p = 0.0330) at 2.5 mg/kg). Eight biomarkers were found, i.e. ß-hydroxyisovaleric acid, leucine, threonine, scylloinositol, lactate, betaine, valine, choline and l-fucose, to be responsible for the class separation between stress and RECA-treated groups. Metabolic pathway alteration in zebrafish brain upon treatment with RECA was identified as valine, leucine and isoleucine biosynthesis, while alanine, aspartate, glutamate and glycerophospholipid metabolism was involved after fluoxetine treatment.