Antioxidant and prooxidant effects of Piptadeniastrum africanum as the possible rationale behind its broad scale application in African ethnomedicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Piptadeniastrum africanum is widely used in treating oxidative stress related diseases. Oxidative stress, defined as the disturbance in the balance between the production of free radicals and antioxidant defenses, is the root cause of many pathophysiological conditions. Based on the dual properties of prooxidants as toxic and beneficial compounds, both prooxidants and antioxidants may be effective in the treatment of these conditions when the right dose is given to the right subject at the right time for the right duration. AIM OF THE STUDY: This study was aimed at investigating the in vitro and ex vivo anti- and pro-oxidative effects of P. africanum. MATERIALS AND METHODS: Total phenolic and flavonoid contents of methanol and aqueous extracts of P. africanum stem back were quantified spectrophotometrically. The methanol extract, ascorbate radicals and reactive oxygen species in brain and liver homogenates of mice treated with the methanol stem bark extract were analyzed by electron paramagnetic resonance (EPR) spectroscopy. Free radical scavenging of DPPH was determined by spectrophotometric and EPR assays. RESULTS: The methanol extract was richer in both phenolic and flavonoid contents compared to the aqueous extracts and also showed better DPPH radical scavenging capacity. The EPR spectroscopy in vitro analysis exhibited high DPPH scavenging capacity before and after UV irradiation (99.5% and 98.76%) at 40 µg/ml extract. The ex vivo EPR spectroscopy studies demonstrated increased levels of ascorbate radicals (•Asc) in liver and brain homogenates of healthy mice treated with P. africanum in comparison with those of the non treated controls (0.6141 ±â€¯0.026 vs 0.1800 ±â€¯0.0073 arb. units for liver homogenates and 0.9605 ±â€¯0.0492 vs 0.3375 ±â€¯0.0062 arb. units for brain homogenates, correspondingly). Considerably, higher levels of reactive oxygen species (ROS) were measured in mice liver and brain homogenates after treatment with P. africanum extract compared to the control group, as well (1.9402 ±â€¯0.1200 vs 0.6699 ±â€¯0.062 arb. units for liver homogenates and 1.7325 ±â€¯01503 vs 0.3167 ±â€¯0.0403 arb.units, respectively). CONCLUSION: Therefore, P. africanum exhibited antioxidant and pro-oxidant properties which may explain its broad spectrum use in a wide variety of ailments.

In vitro studies on radioprotective efficacy of silymarin against γ-irradiation.

UNLABELLED: Abstract Purpose: Silymarin has been widely exploited for its hepatoprotective activities. This study aimed to evaluate the protective efficacy of silymarin against γ-radiation. MATERIALS AND METHODS: The radioprotective properties of silymarin were studied using different assays. Cytotoxicity of silymarin on Human embryonic kidney (HEK) cells was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Protective efficacy against γ-radiation was assessed by studying reduction in micronuclei frequency and free radical generation using 2',7'-dichlorodihydroflurescin diacetate (H2DCFDA). Radiation-induced apoptosis was estimated by Annexin V-PI (propidium iodide) analysis and cell cycle analysis. γ-radiation induced changes in mitochondrial membrane potential (MMP) and DNA damage was estimated employing flow-cytometry and comet assay respectively. RESULTS: MTT assay and Annexin V-PI studies showed that pre-incubation of HEK cells with silymarin protected them from γ-irradiation. Significant reduction in apoptosis (76.36%) was observed. Silymarin also decreased the percentage of radiation-induced micronuclei (> 69%) (p < 0.05 ). Measurement of intracellular reactive oxygen species (ROS) by H2DCFDA revealed a reduction in ROS (21%) at 0.5 h. Cell cycle analysis revealed G1 block in the unirradiated control, which declined in the silymarin pretreated irradiated group (0.5 h). Silymarin treatment resulted in a significant increase in MMP (2 h) against the radiation control. Moreover, the presence of silymarin during irradiation significantly decreased the DNA damage (as measured by comet assay). CONCLUSIONS: Protection against radiation-induced cell-death and DNA damage by silymarin could be attributed to a reduction in ROS induced by γ-radiation. In vitro experiments on HEK cells explicitly prove that silymarin is a promising, effective and safe radiation countermeasure agent.

Antioxidative and radioprotective activities of semiquinone glucoside derivative (SQGD) isolated from Bacillus sp. INM-1.

A semiquinone glucoside derivative (SQGD) was isolated from a radioresistant bacterium Bacillus sp. INM-1 and its antioxidant and radioprotective activities were evaluated using in vitro assays. Natural stable free radical properties of SQGD in solid as well as in solution form were estimated using Electron Paramagnetic Resonance (EPR) spectrometry. Results of the study were demonstrated high reducing power (1.267 ± 0.03356 U(abs)) and nitric oxide radicals scavenging activity (34.684 ± 2.132%) of SQGD. Maximum lipid peroxidation inhibitory activity of SQGD was found to be 74.09 ± 0.08% at 500 µg/ml concentration. Similarly, significant (39.54%; P < 0.05) protection to the liposomal artificial membrane against gamma radiation was observed by SQGD in terms of neutralization of gamma radiation-induced TBARS radicals in vitro. OH(-) radicals scavenging efficacy of SQGD was estimated in terms of % inhibition in deoxy D: -ribose degradation by non-site-specific and site-specific assay. The maximum (54.01 ± 1.01%) inhibition of deoxy D: -ribose degradation was observed in non-site-specific manner, whereas, site-specific inhibition was observed to be 46.36 ± 0.5% at the same concentration (250 µg/ml) of SQGD. EPR spectroscopic analysis of the SQGD indicated ~80% reduction of DPPH radicals at 6.4% concentration. EPR spectral analysis of SQGD was revealed an appearance of very strong EPR signal of 2.00485 (crystalline form) and 2.00520 (solution form) g(y) tensor value, which were an established characteristic of o-semiquinone radicals. Therefore, it can be concluded that SQGD is a natural stable o-semiquinone-type radical, possessing strong antioxidant activities and can effectively neutralize radiation induced free radicals in biological system.