In vivo radioprotective potential of thymol, a monoterpene phenol derivative of cymene.

The radioprotective and anticlastogenic potential of a phenol derivative monoterpene thymol(TOH), against whole-body gamma radiation was studied in Swiss albino mice. Acute toxicity of TOH, with an LD(50(14)) of 1134.03mg/kgbwt., was observed when administered intra-peritoneally (i.p.). The radioprotective potential of TOH was evaluated using the optimal dose of 10mg/kgbwt. TOH, which increased the LD(50/30) by 2.17Gy and resulted in a dose reduction factor (DRF) of 1.25. A significant (p<0.01) reduction in micronucleated, polychromatic erythrocytes (PCE), normochromatic erythrocytes (NCE), and an increased PCE/NCE ratio was also observed after administration of 10mg/kg.b.wt. TOH prior to gamma radiation, indicating its antigenotoxic effect. TOH pre-treatment significantly (p<0.01) elevated reduced glutathione, glutathione-S-transferase, catalase, and superoxide dismutase levels and decreased lipid peroxidation levels in mouse liver homogenates at 24 and 48h after exposure to 4.5Gy of radiation. Further, TOH treatment before exposure to 7.5Gy of gamma radiation resulted in a significant (p<0.01) increase in hematological parameters at various post-treatment time points, with increased numbers of endogenous spleen colonies as well. The histological observations indicated a decline in villus heights and crypt numbers in mouse jejunum and were accompanied by a significant decrease in bone marrow nucleated cells in the irradiated group, which was almost normalized by pre-treatment with TOH. Our study clearly documents the antioxidant, anticlastogenic and radioprotective potentials of TOH, which may be attributed to several possible mechanisms, such as normalization of intracellular antioxidant levels and free radical scavenging activities by TOH.

Mangiferin attenuates methylmercury induced cytotoxicity against IMR-32, human neuroblastoma cells by the inhibition of oxidative stress and free radical scavenging potential.

Mangiferin (MGN), a C-glucosylxanthone was investigated for its ability to protect against methylmercury (MeHg) induced neurotoxicity by employing IMR-32 (human neuroblastoma) cell line. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and clonogenic cell survival assays confirmed the efficacy of MGN supplementation in attenuating MeHg-induced cytotoxicity. Pre-treatment with MGN significantly (p<0.01) inhibited MeHg-induced DNA damage (micronuclei, olive tail moment and % tail DNA) thereby demonstrating MGN's antigenotoxic potential. Also, pre-treatment with MGN significantly reduced MeHg-induced oxidative stress, intra-cellular Ca(2+) influx and inhibited depolarization of mitochondrial membrane. MGN pre-treated cells demonstrated a significant (p<0.05) increase in the GSH and GST levels followed by a significant (p<0.05) decrease in malondialdehyde (MDA) formation. In addition, inhibition of MeHg induced apoptotic cell death by MGN was demonstrated by microscopic, Annexin-V FITC and DNA fragmentation assays and further confirmed by western blot analysis. The present findings indicated the protective effect of MGN against MeHg induced toxicity, which may be attributed to its anti-genotoxic, anti-apoptotic and anti-lipid peroxidative potential plausibly because of its free radical scavenging ability, which reduced the oxidative stress and in turn facilitated the down-regulation of mitochondrial apoptotic signalling pathways.

Mangiferin: A xanthone attenuates mercury chloride induced cytotoxicity and genotoxicity in HepG2 cells.

Mangiferin (MGN), a dietary C-glucosylxanthone present in Mangifera indica, is known to possess a spectrum of beneficial pharmacological properties. This study demonstrates antigenotoxic potential of MGN against mercuric chloride (HgCl2)-induced genotoxicity in HepG2 cell line. Treatment of HepG2 cells with various concentrations of HgCl2 for 3 h caused a dose-dependent increase in micronuclei frequency and elevation in DNA strand breaks (olive tail moment and tail DNA). Pretreatment with MGN significantly (p < 0.01) inhibited HgCl2 -induced (20 µM for 30 h) DNA damage. An optimal antigenotoxic effect of MGN, both in micronuclei and comet assay, was observed at a concentration of 50 µM. Furthermore, HepG2 cells treated with various concentrations of HgCl2 resulted in a dose-dependent increase in the dichlorofluorescein fluorescence, indicating an increase in the generation of reactive oxygen species (ROS). However, MGN by itself failed to generate ROS at a concentration of 50 µM, whereas it could significantly decrease HgCl2 -induced ROS. Our study clearly demonstrates that MGN pretreatment reduced the HgCl2-induced DNA damage in HepG2 cells, thus demonstrating the genoprotective potential of MGN, which is mediated mainly by the inhibition of oxidative stress.

Modulation of gamma ray-induced genotoxic effect by thymol, a monoterpene phenol derivative of cymene.

The radioprotective effect of thymol (TOH), a monoterpene phenol, on radiation-induced DNA damage was analyzed in vitro. Chinese hamster lung fibroblast cells (V79) were treated with different concentrations of TOH (0-100 µg/mL) for 1 hour before exposure to 3 Gy gamma irradiation, and then cytokinesis-blocked micronucleus and single-cell gel electrophoresis (comet assay) assays were used to evaluate the radiation-induced cytogenetic damage and genotoxic effects. Furthermore, the modulating effect of TOH on radiation-induced cell death was assessed by apoptotic and necrotic cell detection by staining with ethidium bromide/acridine orange using fluorescence microscopy. To understand the mechanism of TOH-imparted cytoprotection, mitochondrial membrane potential (MMP) was detected by flow cytometry after staining the cells with Rhodamine 123. Pretreatment of V79 cells with various concentrations of TOH (0-100 µg/mL) for 1 hour reduced the radiation-induced micronuclei as well as percent tail DNA and mean Olive tail moment with a maximum protective effect observed at TOH (25 µg/mL). Apoptosis by microscopic, MMP measurements indicated that the V79 cells exposed to gamma radiation alone showed a maximal increase in the number of early and late apoptotic and necrotic cell death associated with a significant loss of the MMP. Pretreatment with TOH (25 µg/mL) showed a significant (P < .01) decrease in the level of apoptotic fraction as well as necrotic cells and suppressed the radiation-induced collapse of MMP when compared with the radiation alone group. These results suggest that TOH suppresses radiation-induced genotoxicity, apoptosis, and necrosis primarily by the free radical scavenging and modulation of oxidative stress.

Thymol, a naturally occurring monocyclic dietary phenolic compound protects Chinese hamster lung fibroblasts from radiation-induced cytotoxicity.

The effect of thymol (TOH), a dietary compound was investigated for its ability to protect against radiation-induced cytotoxicity in Chinese hamster lung fibroblast (V79) cells growing in vitro. Treatment of V79 cells with 25 microg/ml of TOH prior to 10 Gy gamma radiation resulted increase in the cell viability than that of radiation alone as evaluated by MTT assay. Similarly, there was a significant increase in the surviving fraction observed with 25 microg/ml of TOH administered 1h prior to graded doses of gamma radiation. Further, 25 microg/ml TOH treatment before irradiation significantly decreased the percentage of radiation-induced apoptotic cells (sub-G(1) population) analyzed by flow cytometry as well as DNA ladder assay. TOH was found to inhibit various free radicals generated in vitro, viz., DPPH, O(2), ABTS(+) and OH in a concentration dependent manner. TOH also inhibited the radiation-induced decrease in intracellular glutathione, superoxide dismutase and catalase enzyme levels in V79 cells accompanied by the reduction in lipid peroxides. Our study demonstrated antagonistic potential of TOH against radiation-induced oxidative stress, lipid peroxidation resulting in increased cell viability.

Cytoprotective and antigenotoxic potential of Mangiferin, a glucosylxanthone against cadmium chloride induced toxicity in HepG2 cells.

Mangiferin (MGN), a glucosylxanthone present in large amounts in the leaves and edible mango fruits of Mangifera indica. Here, we report about MGN's potential for mitigating cadmium chloride (CdCl(2)) induced cytotoxic and genotoxic effects in HepG2 cells growing in vitro. The cytoprotective potential was assessed by MTT, clonogenic and apoptotic assays, while antigenotoxic effect by micronucleus and comet assay. The established cytotoxic and genotoxic effects were well indicated after CdCl(2) treatment and was mitigated by pretreatment with MGN. MGN prior to CdCl(2) treatment increased the cell survival (MTT), surviving fraction (clonogenic assay) and inhibited sub-G(1) population (flow cytometric analysis). Further, inhibition of CdCl(2) induced apoptotic cell death by MGN was confirmed by microscopic and DNA fragmentation assays. A significant (p<0.01) reduction in the micronuclei frequency and comet parameters after MGN pretreatment to CdCl(2) clearly indicated the antigenotoxic potential. Similarly, the reactive oxygen species generated by the CdCl(2) treatment were inhibited significantly (p<0.001) by MGN. Taken together, our study revealed that MGN has potent cytoprotective and antigenotoxic effect against CdCl(2) induced toxicity in HepG2 cell line and which may be attributed to decrease in CdCl(2) induced reactive oxygen species levels and resultant oxidative stress.