Effect of diosmin on apoptotic signaling molecules in N-nitrosodiethylamine-induced hepatocellular carcinoma in experimental rats.

The aim of the present study was to evaluate the antioxidant and chemopreventive efficiency of diosmin against N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in adult male rats. Rats were classified into four groups as follows: Group I: Control, Group II: NDEA-induced hepatocellular carcinogenic rats, Group III: Cancer-bearing animals treated with diosmin (200 mg/kg/body weight/day) orally for 28 days, Group IV: Control animals treated with diosmin (200 mg/kg/body weight/day) alone for 28 days. The model of NDEA-induced HCC rats elicited significant increases in alpha-fetoprotein (AFP), lipid peroxidation (LPO) and increase in anti-apoptotic proteins (Bcl-2, Bcl-xL and Mcl-1) with a concomitant significant decline in liver antioxidant enzymes, pro-apoptotic (Bax and Bad) and caspase-3 &-9 proteins. The oral administration of diosmin as a protective agent normalized the altered levels of AFP, LPO, antioxidant enzymes, pro- and anti-apoptotic proteins as well as caspase-3 and -9 proteins. Transmission electron microscopical studies also revealed that treatment of diosmin has a perspective anti-cancer activity by rearranging hepatic cell structure and its integrity. Results of this study suggest that diosmin may be one of a pharmacological and therapeutic representative against hepatocellular carcinoma.

Modulating effects of hesperidin on key carbohydrate-metabolizing enzymes, lipid profile, and membrane-bound adenosine triphosphatases against 7,12-dimethylbenz(a)anthracene-induced breast carcinogenesis.

The aim of this study was to document the effect of hesperidin on the key enzyme activities of carbohydrate metabolism, lipid profile, and membrane-bound adenosine triphosphatases (ATPases) during 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast carcinogenesis. Hesperidin has been reported to have multiple biological properties. Breast cancer was induced by single dose of DMBA (20 mg/kg body weight (bw)). The results revealed that there was a significant increase in the activities of hexokinase, phosphoglucoisomerase, and aldolase and a concomitant decrease in the activities of glucose-6-phosphatase and fructose-1,6-diphosphatase in cancer-induced animals. The activities of ATPases were found to be decreased both in erythrocyte membrane and in the liver of mammary cancer-bearing animals. The lipid profiles such as total cholesterol, free cholesterol, phospholipids, triglycerides, and free fatty acids significantly increased and in contrast the ester cholesterol in plasma was found to be decreased, whereas it was found to be elevated in the liver of cancer-bearing groups. The altered levels of the above-mentioned biochemical parameters in cancer-bearing animals were significantly ameliorated by the administration of hesperidin at the dosage of 30 mg/kg bw for 45 days. The histopathological analysis of breast and liver tissues were well supported the modulatory property of hesperidin, and this might be associated with normalizing the gluconeogenesis process, stabilization of cell membranes, and modulation of lipid biosynthesis.

Influence of limonin on Wnt signalling molecule in HepG2 cell lines.

OBJECTIVE: The role of limonin as potent anti carcinogenic, apoptosis and chemotherapeutic agents has been supported by limited studies. MATERIALS AND METHODS: In this study, limonin is identified as a potent anti proliferative agent against human hepatoma HepG2 cells based on the cell viability study, LDH leakage assay. Induction of apoptosis in HepG2 cells by limonin was evidenced by western blot analysis of Bax, Cyclin D1, Caspase 3 and Caspase9. RESULTS: Since Wnt signalling is involved in the initiation and sustaining of hepatocellular carcinoma we studied differential expression of LRP5, LRP6 and DKK wnt players. CONCLUSION: Limonin found to down regulate these players which forms a rationale for further investigation on effect on limonin in cancer therapy.

Inhibitory effects of Indigofera aspalathoides on 20-methylcholanthrene-induced chemical carcinogenesis in rats.

BACKGROUND: The anticancer and antioxidant effects of the aqueous extract of Indigofera aspalathoides on 20-methylcholanthrene (20-MCA) induced fibrosarcoma were investigated in male albino rats. MATERIALS AND METHODS: The rats were divided into four different groups, each group consisting of six animals. Group I animals were served as normal control, Group II animals were fibrosarcoma-bearing animals after the incubation period, Group III animals were fibrosarcoma-bearing animals, treated with aqueous extract of I. aspalathoides intraperitoneally at a dose of 250 mg/kg b.w. for 30 days and Group IV animals were administered with the aqueous extract of I. aspalathoides alone, at a dose of 250 mg/kg b.w. for 30 days, served as drug control animals. After the experimental period, all the rats were weighed and killed by cervical decapitation. The serum was separated from the blood for analysis. The weights of the liver and the kidneys were noted. The fibrosarcoma was proved by pathological examinations. The liver and kidney tissues were excised and then homogenized in an ice-cold buffer. These tissues were used for biochemical analysis. RESULTS: The activities of antioxidant enzymes, e.g. catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD), in blood serum, liver, and kidney of control and experimental animals, respectively, have been reported. CONCLUSION: The present observations suggested that the aqueous extract of I. aspalathoides treatment enhanced the recovery from 20-MCA-induced fibrosarcoma due to its antioxidants and antineoplastic properties.

Indigofera aspalathoides protection against 20-methylcholanthrene-induced experimental fibrosarcoma growth after transplantation in rats - role of xenobiotic drug metabolizing enzymes.

A large number of active principles from traditional medicinal plants have been reported to have chemopreventive properties. In the present study, therapeutic efficacy of an aqueous extract of Indigofera aspalathoides against growth of transplanted experimental fibrosarcomas in Wistar strain male albino rats was tested. Tumors which appeared about six weeks after implantation were highly localized and were maintained by serial transplantation. Rats were divided into four groups. Group I served as normal control animals. Group II were fibrosarcoma bearing animals. Group III were animals with fibrosarcoma treated with Indigofera aspalathoides aqueous extracts at a dose of 250 mg/kg. b. w. per day for 30 days. Group IV animals were treated with aqueous extract of Indigofera aspalathoides alone. Reduction in tumor weight was noted in Group III as compared to II. The levels of cytochrome C in liver and kidney, the levels of cytochrome P450 and cytochrome b5 in liver microsomes, phase I biotransformation enzymes NADPH-cytochrome P450, NADPH-cytochrome b5, and aniline hydroxylase, and the phase II enzymes glutathione-S-transferase and UDP glucuronyl transferase indicated that their modulation played a role in the therapeutic efficacy of Indigofera aspalathoides against experimental fibrosarcoma.

DNA damage and cell necrosis induced by naphthalene due to the modulation of biotransformation enzymes in an estuarine crab Scylla serrata.

The sublethal effect of naphthalene (2.5, 5, and 10 mg L(-1)) was studied in an estuarine crab Scylla serrata with reference to macromolecular changes. Biotransformation enzymes such as cytochrome P450, cytochrome b(5), NADPH cytochrome P450 reductase, aryl hydrocarbon hydroxylase, glutathione-S-transferase, and UDP-glucuronyl transferase were elevated in the hepatopancreas of naphthalene-exposed crabs in comparison with control. Remarkable amount of DNA damage and cell necrosis was observed in hepatopancreas, hemolymph, and ovary of the crabs exposed to naphthalene, when compared with control. For all the parameters studied, a concentration-dependent gradient of the changes was observed. The expression of DNA damage and cell necrosis suggests an increased production of oxidants during naphthalene metabolism.

Reproductive dysfunction induced by naphthalene in an estuarine crab Scylla serrata with reference to vitellogenesis.

Biomarkers are useful tools for understanding complex interactions that elicit organisms response to environmental pollutants and their sublethal effects on organisms health. Effect of naphthalene on vitellogenin (VTG) and vitellin (VTN) were assessed in hepatopancreas, haemolymph and ovary of an estuarine crab Scylla serrata with reference to vitellogenic phases. In addition, the gonadosomatic index (GSI) was also assessed. Significant reductions in VTG and VTN contents were observed in hepatopancreas, haemolymph (VTG) and ovary (VTN). The GSI exhibited a decreasing trend in crabs exposed to naphthalene irrespective of the vitellogenic phases. We attempted to use the alterations in vitellogenic proteins and GSI as biomarkers of reproductive disturbances occurred in the crab due to naphthalene stress.

Sublethal effect of silver and chromium in the green mussel Perna viridis with reference to alterations in oxygen uptake, filtration rate and membrane bound ATPase system as biomarkers.

Perna viridis is an ideal animal for studying the impairment caused by the effects of heavy metals that are often detected in coastal areas. Preliminary bioassay tests revealed that the lethal (LC(100)), median lethal (LC(50)) and sublethal (LC(0)) concentration of silver and chromium to P. viridis were 6.5, 4.0, 2.0 mg l(-1) and 4.5, 2.5, 1.0 mg l(-1), respectively. Toxic effect of silver and chromium was evaluated in the green mussel P. viridis, with reference to oxygen consumption, filtration rate and ATPase system in laboratory experiments. These parameters were selected as the end point of sublethal stress. Oxygen consumption and filtration rates were calculated as a measure of decline in the dissolved oxygen level and algal concentration (feed) in the aquaria water, respectively. Silver and chromium affects both oxygen consumption and filtration rate significantly (P<0.01) at 96 h when compared to control. The activity of ATPases system in the gills, hepatopancreas, ovary and muscle of mussels were inhibited by silver and chromium indicating that metals exerted significant toxic effect. The inhibition of Na(+)K(+) ATPase, Ca(2+) ATPase and Mg(2+) ATPase in the mussels were significant (P<0.05) for silver and highly significant (P<0.01) for chromium, which indicates that chromium was more toxic to mussels when compared to silver. The assessment of oxygen consumption, filtration and ATPases system can thus be used as a valid biomarker in aquatic ecotoxicology studies.

Protective effect of Operculina turpethum against 7,12-dimethyl benz(a)anthracene induced oxidative stress with reference to breast cancer in experimental rats.

Reactive oxygen species (ROS) directly or indirectly involves in multistage process of carcinogenesis. Antioxidant activity of methanolic extract of Operculina turpethum stems (MEOT) on 7,12 dimethylbenz(a)anthracene (DMBA) induced breast cancer was investigated in female Sprague-Dawley rats. Changes in the levels of lipid peroxidation and antioxidants system was evaluated in addition to tumour development. Twenty four female rats were divided into four groups: control, DMBA, DMBA+MEOT and MEOT. In the DMBA group, rats were intragastrically administered with 20 mg of DMBA using corn oil as vehicle. Animals of DMBA+MEOT group received a single dose of 20 mg of DMBA dissolved in corn oil intragastrically followed by O. turpethum extract (100 mg/kg body weight), while MEOT group received O. turpethum extract (100 mg/kg body weight) intragastrically daily for a period of 45 days. After the experimental period of 45 days, oxidative stress parameters were assessed in serum, liver and breast of both control and experimental groups. In addition to this, tumour weight of breast was also assessed. A significant increase in lipid peroxidation levels were observed in the tested samples of cancer induced rats while the activities of enzymic antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and non-enzymic antioxidants like glutathione (GSH), ascorbic acid (Vitamin C) and alpha-tocopherol (Vitamin E) were decreased in cancer-bearing animals when compared to control animals. A significant (P<0.05) increase in the tumour weight was observed in the breast of DMBA group and the breast tumour weight decreased significantly (P<0.05) in the DMBA+MEOT groups. Oral administration of MEOT remarkably reduced the lipid peroxidation activity and increased the antioxidants level in drug treated animals and decreased the tumour weight significantly (P<0.05). This result suggests that MEOT shows antioxidant activity and play a protective role against DMBA induced breast cancer.

Antioxidant effect of the marine algae Chlorella vulgaris against naphthalene-induced oxidative stress in the albino rats.

Alcoholic extract of the marine algae Chlorella vulgaris was examined for its free radical scavenging effect with reference to naphthalene-induced lipid peroxidation in serum, liver, and kidney of rats. Initially, upon naphthalene intoxication (435 mg/kg body weight, intraperitoneally), the lipid peroxidation activity increased significantly (P < 0.001), and in contrast, the enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase) and non-enzymic antioxidants (glutathione, ascorbic acid, and alpha-tocopherol) levels decreased remarkably. When the naphthalene stressed rats were treated with Chlorella vulgaris extract (70 mg/kg body weight, orally), the lipid peroxidation activity reduced significantly (P < 0.001) and the activities of both the enzymic and non-enzymic antioxidants increased reaching near control values. The minimum concentration (70 mg/l) of the extract that exhibited maximum (85%) free radical scavenging activity was chosen for the experimental study. The present results suggest that Chlorella vulgaris extract exerts its chemo-preventive effect by modulating the antioxidants status and lipid peroxidation during naphthalene intoxication.

Interaction of potash and decis in the ecophysiology of a freshwater fish Oreochromis mossambicus.

Interaction of potash and decis in the ecophysiology of a freshwater fish, Oreochromis mossambicus, was studied. It was found that 300, 550 and 700 mgL(-1) of potash were sublethal (LC(0)), median lethal (LC(50)), and toxic (LC(100)) to O. mossambicus for 96h exposure, respectively. For decis, 96 h LC(100,) LC(50), and LC(0) was 0.4, 0.25, and 0.1 mgL(-1), respectively. Sublethal concentrations of potash and decis were exposed to fishes individually and in combination for 28 days. The results revealed that the combined effect of these chemicals was more highly toxic to food intake, growth, and conversion efficiencies than the individual chemicals. The marker enzymes (acid phosphatase, alkaline phosphatase, aspartate transaminase, alanine transaminase) were also analyzed in blood, liver and muscle. The enzyme activities were decreased in liver and muscle. On the other hand, serum exhibited increased activities of marker enzymes. The results were tested statistically and interpreted accordingly.

Dietary ascorbic acid and alpha-tocopherol mitigates oxidative stress induced by copper in the thornfish Terapon jarbua.

The protective effects of vitamins C and E against oxidative stress were evaluated in various tissues of thornfish Terapon jarbua exposed to copper. Preliminary bioassay tests performed with copper and T. jarbua revealed that 4.0, 2.5 and 1.0 mg L(-1) of copper were lethal (LC(100)), medial lethal (LC(50)) and sublethal (LC(0)) respectively. Oxidative stress parameters viz. lipid peroxidation, superoxide dismutase, catalase, glutathione peroxidase, and glutathione were evaluated in control and experimental fishes. Lipid peroxidation activity increased in tissues of copper exposed fishes, while the antioxidant system exhibited a reduction in their activity. On the contrary copper stressed fishes fed with vitamins C and E enriched pellet feed showed significant reduction in lipid peroxidation activity and the antioxidant levels increased reaching near normal levels comparable to control values. Bioaccumulation of copper was studied in addition to oxidative stress. Substantial copper residue was detected in the tissues of T. jarbua exposed to copper and the level of copper in tissues reduced when the fishes were treated with vitamins ensuring copper depuration and thereby protecting them against stress. We concluded that vitamin supplementation offered significant reduction of the oxidative stress mediated by copper and we discuss the possible application of vitamins in costal aquaculture process.

Free radical scavenging activity of the marine mangrove Rhizophora apiculata bark extract with reference to naphthalene induced mitochondrial dysfunction.

Rhizophora apiculata bark extract was tested for its free radical scavenging activity and protective role against mitochondrial dysfunction in naphthalene stressed rats. Lipid peroxidation activity was increased and activity of mitochondrial enzymes (cytochrome-c-oxidase, NADH-dehydrogenase, alpha-ketoglutarate dehydrogenase and succinate dehydrogenase) and glutathione was decreased in the liver and kidney of rats intoxicated with naphthalene when compared to control rats. Intraperitoneal administration of plant extract significantly reduced the lipid peroxidation, increased the activity of mitochondrial enzymes and increased glutathione to near control levels. These results suggest that the sulfated polysaccharides in R. apiculata play a protective role through their free radical scavenging properties.

Assessment of biochemical components and enzyme activities in the estuarine crab Scylla tranquebarica from naphthalene contaminated habitants.

To document the impact of naphthalene, a comparative toxicological research was performed in the estuarine crab Scylla tranquebarica habitant of Ennore creek (polluted site) and Kovalam creek (pristine site) for a period of 6 months. The biochemical constituents, such as protein, carbohydrate, lipid and enzyme activities like acid and alkaline phosphatase, aspartate and alanine transaminase were analysed in hepatopancreas, heamolymph and ovary of the female crabs collected from the two creeks. The results revealed that there was a significant measurable difference in these parameters in the tissues of crabs sampled from the polyaromatic hydrocarbon (PAH) polluted site (Ennore creek) when compared with the reference site (Kovalam creek). In addition naphthalene was detected in gill, hepatopancrease, heamolymph and ovary of the crabs sampled from the polluted creek. The results indicate that the indicators selected for toxic impact in S. tranquebarica may be related to the uptake of naphthalene in the tissues examined and support the feasibility of employing these types of analyses as biochemical biomarkers for PAH contaminant-exposed organisms.

Fluctuations of biochemical constituents and marker enzymes as a consequence of naphthalene toxicity in the edible estuarine crab Scylla serrata.

The sublethal effects of naphthalene on protein, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), carbohydrates, lipids, and certain marker enzymes such as phosphatases, transaminases, and lactate dehydrogenase were studied in hepatopancreas, hemolymph, and ovary in the edible crab Scylla serrata. The results revealed that there was overall decrease in total protein, total DNA, total RNA, free sugar, glycogen, protein-bound sugars, neutral lipid, glycolipid, and phospholipid in the test samples compared to control. Similarly all the marker enzymes (acid phosphatase, alkaline phosphatase, aspartate transaminase, alanine transaminase, and lactate dehydrogenase) were decreased in hepatopancreas and ovary. On the other hand, in hemolymph, the activities of marker enzymes were increased. The results were tested statistically and interpreted accordingly.

Changes in oxygen consumption and respiratory enzymes as stress indicators in an estuarine edible crab Scylla serrata exposed to naphthalene.

The sublethal effect of naphthalene was studied on the physiology of a mud crab Scylla serrata. The 96 h acute toxicity of naphthalene was determined and found to be 28 mg 1(-1) (LC100), 18 mg 1(-1) (LC50), 10 mg 1(-1) (LC0) respectively. The 30 days sublethal effect (LC0) 9 mg 1(-1), 8 mg 1(-1), 10 mg 1(-1), of naphthalene was investigated in the crab S. serrata with reference to oxygen consumption and changes in the activity of respiratory enzymes. The results indicated that naphthalene caused disturbance in the normal physiology of the crab. The bioaccumulation of naphthalene was also investigated in gills, hepatopancreas, haemolymph and ovary. The consumption of oxygen increased in the naphthalene medium when compared with that of the crabs exposed to naphthalene free medium. A decreased trend in the activity of respiratory enzymes such as lactate dehydrogenase (LDH), isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), alpha-ketoglutarate dehydrogenase (alpha-KDH) and glutathione (GSH) were recorded in the hepatopancreas, ovary and gills of S. serrata for all the tested concentrations of naphthalene and the results were analyzed for their significance.

Biochemical changes in serum, lung, heart and spleen tissues of mice exposed to sub-acute toxic inhalation of mosquito repellent mat vapour.

The toxicity of sub-acute inhalation of mosquito repellent mat vapour exposure 3 hr daily, either for a duration of 15 days (Group II) or for 30 days (Group III) on biochemical parameters in serum and vital organs was evaluated in mice. The control animals were exposed to ambient air 3 hr daily, either for 15 days [Group I (a)] or 30 days [Group I (b)]. In both the Group II and Group III exposure studies, elevation in the activity of aspartate transaminase (AST) and alanine transaminase (ALT) was observed in the serum and heart tissue and a decrease in their activity in lung and spleen tissue. This biochemical lesion indicates that exposure to mat vapour is capable of interfering with the metabolism of non-essential amino acids and induce cellular damage in the tissues. The elevation in the activity of AST along with that of lactate dehydrogenase (LDH) observed in serum and heart tissue in Group II and III studies indicate that exposure to mat vapour is capable of inducing changes in the myocardial function. The elevation in the activity of alkaline phosphatase (ALP) in serum and a decrease in its activity in lung and heart tissue of mice exposed to mat vapour for 30 days (Group III) observed in this study denotes that mat vapour could interfere with various metabolic functions to induce derangements at cellular level. Inhalation of hitherto unidentified volatile chemicals emanated from mosquito repellent mat, may contribute as stress factors may contribute as in an indoor environment to produce above mentioned biochemical lesions in vital organs.

Influence of insecticidal derivative (cartap hydrochloride) from the marine polycheate on certain enzyme systems of the fresh water fish Oreochromis mossambicus.

The activities of phosphatases and transaminases were studied in muscle and liver of the fresh water fish, Oreochromis mossambicus on exposure to different sublethal concentrations (0.25, 0.5, 0.75 and 1 mgl(-1)) of cartap hydrochloride (insecticidal derivative from marine polycheate) for 96 h. There was an overall decrease in phosphatases and transaminases activity in muscle and liver of the fish subjected to cartap hydrochloride.

Impact of distillery effluent on carbohydrate metabolism of freshwater fish, Cyprinus carpio.

The impact of distillery effluent on carbohydrate metabolism of Cyprinus carpio was studied at different days during exposure (7, 14, and 21 days) in the ambient temperature of 28 +/- 1 degrees C. Oxygen consumption in fish decreased with increasing effluent concentrations as well as duration of exposure. Effluent concentrations and exposure durations (days) had a significant effect on oxygen consumption of tested fish (p < 0.0005). Total carbohydrate, glycogen content and SDH enzyme activity in muscle, liver and brain tissues of C. carpio exposed to different sublethal concentrations decreased gradually and significantly. This was also the case with exposure duration. Reduction in glycogen content was greater in liver tissue i.e., 54.1% in 0.2% effluent concentration on the 21st day of exposure. However, serum glucose and lactic acid content showed an increasing trend with increase in effluent concentration and time of exposure. Unlike SDH, LDH enzyme activity of muscle, liver and brain tissues showed an increasing trend and the enhancement of enzyme activity was more in liver tissue (71.3%). From these results, it could be inferred that respiratory processes in C. carpio under distillery effluent stress was affected resulting in a shift towards anaerobiosis at organ level during sublethal intoxication.

Naphthalene-induced hematological disturbances and oxidative stress in an estuarine edible crab, Scylla serrata.

The 30-day sublethal effect of naphthalene on Scylla serrata was studied using hematological parameters, lipid peroxidation, and antioxidant status. The results revealed a decreasing trend in the hemolymph-related parameters of the crabs exposed to naphthalene. There was an increase in lipid peroxidation activity in the gonads of the crabs exposed to naphthalene. On the other hand, the enzymatic (superoxide dismutase, catalase, glutathione peroxidase) and nonenzymatic (vitamin E, vitamin C and glutathione) antioxidants showed decreased activity for the respective gonads. The results were statistically significant in comparison with the control. These parameters are considered the first detectable/quantifiable response to chemical stress and can serve as biomarkers.