Reversal of cadmium chloride-induced oxidative stress and genotoxicity by Adhatoda vasica extract in Swiss albino mice

Adhatoda vasica Nees (Acanthaceae) that is used by Ayurvedic physicians possesses some established medicinal properties. Environmental and occupational exposure with cadmium affects the renal system adversely. Cadmium is an established genotoxic agent. In the present study, we evaluated the antioxidant and anticlastogenic efficacy of A. vasica against cadmium chloride (CdCl₂)-induced renal oxidative stress and genotoxicity in Swiss albino mice. A single intraperitoneal dose of CdCl₂ (5 mg/kg BW) resulted in significant (p<0.001) increase in chromosomal aberration and micronuclei formation. Oral administration of A. vasica at two doses (50 and 100 mg/kg BW) for seven consecutive days showed significant (p<0.001) suppression of mutagenic effects of CdCl₂ in plant-pretreated groups. To study the mechanism by which A. vasica exerts its antimutagenic potential, enzymes involved in metabolism and detoxification were also estimated. Cadmium intoxication altered the antioxidant levels and enhanced MDA formation significantly (p<0.001). A. vasica showed significant (p<0.001) recovery in antioxidant status, viz., GSH content, its dependent enzymes, and catalase activity. Prophylactic pretreatment of A. vasica extract in cadmium-intoxicated mice showed marked (p<0.001) inhibition of lipid peroxidation (LPO) and xanthine oxidase (XO) activity. The present findings support that antimutagenic efficacy of A. vasica can be attributed to its restoring effects on antioxidant status and suppression of MDA level formation.     

Modulatory role of Emblica officinalis fruit extract against arsenic induced oxidative stress in Swiss albino mice

Arsenic, an important human toxin, is naturally occurring in groundwater and its accumulation in plants and animals have assumed a menacing proportion in a large part of the world, particularly Asia. Epidemiological studies have shown a strong association between chronic arsenic exposure and various adverse health effects, including cardiovascular diseases, neurological defects and cancer of lung, skin, bladder, liver and kidney. The protective role of the fruits of Emblica officinalis (500 mg/kg b.wt.) was studied in adult Swiss albino mice against arsenic induced hepatopathy. Arsenic treated group (NaAsO₂, 4 mg/kg b.wt.) had a significant increase in serum transaminases and lipid peroxidation (LPO) content in liver, whereas significant decrease was recorded in hepatic superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) and serum alkaline phosphatase activity. Combined treatment of Emblica and arsenic (pre and post) declined the serum transaminases and LPO content in liver whereas significant increase was noticed in SOD, CAT, GST and serum alkaline phosphatase activities. Liver histopathology showed that Emblica fruit extract had reduced karyolysis, karyorrhexis, necrosis and cytoplasmic vacuolization induced by NaAsO₂ intoxication. Thus it can be concluded that pre- and post-supplementation of E. officinalis fruit extract significantly reduced arsenic induced oxidative stress in liver.     

Inhibition of cutaneous oxidative stress and two-stage skin carcinogenesis by Hemidesmus indicus (L.) in Swiss albino mice

Chemopreventive potential of H. indicus on 7,12-dimethyl-benz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoyl 13-phorbol acetate (TPA) promoted murine skin carcinogenesis has been assessed. Topical application of H. indicus resulted in significant protection against cutaneous tumorigenesis. Topical application of plant extract prior to that of TPA resulted in significant inhibition against TPA-caused induction of epidermal ornithine decarboxylase (ODC) activity and DNA synthesis. Application of H. indicus at a dose level of 1.5 and 3.0 mg/kg body weight in acetone prior to that of TPA treatment resulted in significant inhibition of oxidative stress. The level of lipid peroxidation was significantly reduced. In addition, depleted levels of glutathione and reduced activities of antioxidant enzymes were restored respectively). The results indicate that H. indicus is a potent chemopreventive agent in skin carcinogenesis.     

Glycyrrhizin exhibits potential chemopreventive activity on 12-O-tetradecanoyl phorbol-13-acetate-induced cutaneous oxidative stress and tumor promotion in Swiss albino mice

Glycyrrhizin and its aglycone, glycyrrhetic acid has been found useful for various therapeutic purposes. Glycyrrhizin has been shown to possess many physiological functions like anti-inflammatory activity, detoxification and inhibition of carcinogenic promoters. 12-O-Tetradecanoyl phorbol-13-acetate (TPA), a well-known phorbal ester is known for its tumor promotion activity. The induction of inflammation in skin mediated by TPA is believed to be governed by cyclooxygenase (COX), lipoxygenase and ornithine decarboxylase (ODC). These markers of inflammatory responses are important for skin tumor promotion. In our present study, we studied the chemopreventive effect of glycyrrhizin on TPA (20 nmol/0.2 mL acetone/animal, topically)-induced oxidative stress and hyperproliferation markers in skin. TPA enhanced lipid peroxidation with reduction in the level of catalase, glutathione, glutathione peroxidase, glutathione reductase and glutathione-s-transferase. TPA treatment also enhanced ODC activity and [3H] thymidine incorporation into cutaneous DNA. Prophylactic treatment of mice with glycyrrhizin (2.0 & 4.0 mg/0.2 mL acetone/animal, topically) resulted in a significant decrease in cutaneous microsomal lipid peroxidation (P < 0.001) and recovery of cutaneous glutathione content (P < 0.001) and its dependent enzymes. A significant inhibition in ODC activity and DNA synthesis (P < 0.001) was also observed. Thus, the results demonstrate that pretreatment with glycyrrhizin is protective against TPA-induced oxidative stress and tumor promotion in Swiss albino mice.     

The in vivo genotoxicity of cisplatin,isoflurane and halothane evaluated by alkaline comet assay in Swiss albino mice

The aim of this study was to evaluate the genotoxicity of repeated exposure to isoflurane or halothane and compare it with the genotoxicity of repeated exposure to cisplatin. We also determined the genotoxicity of combined treatment with inhalation anaesthetics and cisplatin on peripheral blood leucocytes (PBL), brain, liver and kidney cells of mice. The mice were divided into six groups as follows: control, cisplatin, isoflurane, cisplatin–isoflurane, halothane and cisplatin–halothane, and were exposed respectively for three consecutive days. The mice were treated with cisplatin or exposed to inhalation anaesthetic; the combined groups were exposed to inhalation anaesthetic after treatment with cisplatin. The alkaline comet assay was performed. All drugs had a strong genotoxicity (P < 0.05 vs. control group) in all of the observed cells. Isoflurane caused stronger DNA damage on the PBL and kidney cells, in contrast to halothane, which had stronger genotoxicity on brain and liver cells. The combination of cisplatin and isoflurane induced lower genotoxicity on PBL than isoflurane alone (P < 0.05). Halothane had the strongest effect on brain cells, but in the combined treatment with cisplatin, the effect decreased to the level of cisplatin alone. Halothane also induced the strongest DNA damage of the liver cells, while the combination with cisplatin increased its genotoxicity even more. The genotoxicity of cisplatin and isoflurane on kidney cells were nearly at the same level, but halothane caused a significantly lower effect. The combinations of inhalation anaesthetics with cisplatin had stronger effects on kidney cells than inhalation anaesthetics alone. The observed drugs and their combinations induced strong genotoxicity on all of the mentioned cells.     

Alleviation of free radical mediated oxidative and genotoxic effects of cadmium by farnesol in Swiss albino mice

Abstract

Farnesol is an isoprenoid found in essential oils of ambrette seeds, citronella and in various aromatic plants. Exposure to cadmium from various sources affects the renal system adversely and Cd is an established genotoxic agent. In the present study, we evaluated the antigenotoxic and antioxidant efficacy of farnesol against cadmium chloride (CdCl₂)-induced renal oxidative stress and genotoxicity in Swiss albino mice. Single, intraperitoneal doses of CdCl₂(5 mg/kg body weight) for 24 h resulted in a significant (P < 0.001) increase in chromosomal aberration and micronuclei formation. The oral administration of farnesol at two doses (1% and 2% per kg body weight) for seven consecutive days showed significant (P < 0.05) suppression of the genotoxic effects of CdCl₂ in the modulator groups. To study the mechanism by which farnesol exerts its antigenotoxic potential, enzymes involved in metabolism and detoxification were estimated. CdCl₂ intoxication adversely affected the renal antioxidant armory and increased TBARS formation and xanthine oxidase levels significantly (P < 0.001). Farnesol showed a significant (P < 0.001) recovery in antioxidant status viz, GSH content (and its dependent enzymes) and catalase activity. Farnesol pretreatment in CdCl₂-intoxicated mice showed marked (P < 0.001) suppression of TBARS' formation and XO activity. Our results support the conclusion that the anticlastogenic effect of farnesol could be due to restoration of antioxidants and inhibition of oxidative damage.     

Alleviation of free radical mediated oxidative and genotoxic effects of cadmium by farnesol in Swiss albino mice

Farnesol is an isoprenoid found in essential oils of ambrette seeds, citronella and in various aromatic plants. Exposure to cadmium from various sources affects the renal system adversely and Cd is an established genotoxic agent. In the present study, we evaluated the antigenotoxic and antioxidant efficacy of farnesol against cadmium chloride (CdCl2)-induced renal oxidative stress and genotoxicity in Swiss albino mice. Single, intraperitoneal doses of CdCl2(5 mg/kg body weight) for 24 h resulted in a significant (P < 0.001) increase in chromosomal aberration and micronuclei formation. The oral administration of farnesol at two doses (1% and 2% per kg body weight) for seven consecutive days showed significant (P < 0.05) suppression of the genotoxic effects of CdCl2 in the modulator groups. To study the mechanism by which farnesol exerts its antigenotoxic potential, enzymes involved in metabolism and detoxification were estimated. CdCl2 intoxication adversely affected the renal antioxidant armory and increased TBARS formation and xanthine oxidase levels significantly (P < 0.001). Farnesol showed a significant (P < 0.001) recovery in antioxidant status viz, GSH content (and its dependent enzymes) and catalase activity. Farnesol pretreatment in CdCl2-intoxicated mice showed marked (P < 0.001) suppression of TBARS' formation and XO activity. Our results support the conclusion that the anticlastogenic effect of farnesol could be due to restoration of antioxidants and inhibition of oxidative damage.     

Marked variation in diazepam sensitivity in Swiss albino mice

Using the righting reflex as the critical level, sleep was measured in Swiss albino mice at a dose of 35 mg/kg diazepam, i.p. Sleep times varied markedly from zero to 120 min with a mean +/- s.d. of 44 +/- 37 (N = 202). The distribution is skewed to the left with a coefficient of skewness of 0.33 +/- 0.17. The sleep times of the two sexes, when analyzed separately, showed similar range, mean and s.d., except that the distribution tended to be more clearly bimodal in males than in females. These animals also exhibited marked variations in their response to either ethanol (4 g/kg) or pentobarbital (45 mg/kg). The diazepam sleep time failed to correlate with the ethanol sleep time. Significant correlation, however, was obtained between diazepam and pentobarbital sleep times. On further analysis with least-squares fit to a straight line, the data yielded a line with a slope of 0.16; thus despite the correlation reaching a significant level, there is no significant difference in the pentobarbital sleep times between mice that have the longest or the shortest diazepam sleep times. By monitoring the plasma and brain levels of diazepam and N-desmethyldiazepam in mice at awakening, it was found that the variations in sleep time cannot be explained by individual differences in drug disposition. The phenomenon is discussed in terms of individual variations in diazepam sensitivity and the possibility of development of tolerance to diazepam almost immediately after diazepam administration.     

Cyclophosphamide-induced nephrotoxicity, genotoxicity, and damage in kidney genomic DNA of Swiss albino mice: the protective effect of Ellagic acid

Cyclophosphamide (CPM), an alkylating agent is used as an immunosuppressant in rheumatoid arthritis and in the treatment of several cancers as well. In this study, Ellagic acid (EA), a naturally occurring plant polyphenol, was evaluated for its antigenotoxicity and antioxidant efficacy against the CPM-induced renal oxidative stress and genotoxicity in Swiss albino mice. The mice were given a prophylactic treatment of EA orally at a dose of 50 and 100 mg/kg body weight (b wt) for seven consecutive days before the administration of a single intraperitoneal (i.p.) injection of CPM at 50 mg/kg b wt. The modulatory effects of EA on CPM-induced nephrotoxicity and genotoxicity were investigated by assaying oxidative stress biomarkers, serum kidney toxicity markers, DNA fragmentation, alkaline unwinding assay, micronuclei (MN) assay, and by histopathological examination of kidney tissue. A single intraperitoneal administration of CPM in mice increased malondialdehyde level with depletion in glutathione content, antioxidant enzymes activities, viz. glutathione peroxidase, glutathione reductase, catalase, quinone reductase, induced DNA strand breaks, and MN induction. EA oral administration at both doses caused significant reduction in their levels, restoration in the activities of antioxidant enzymes, reduction in MN formation, and DNA fragmentation. Serum toxicity marker enzymes like BUN, creatinine, and LDH were also increased after CPM treatment which was significantly decreased in EA pretreated groups. Present findings suggest a prominent role of EA against CPM-induced renal injury, DNA damage, and genotoxicity.     

Effect of sesamol on radiation-induced cytotoxicity in Swiss albino mice

The radio-protective ability of sesamol (SM) at various doses viz., 0, 10, 25, 40, 50, 70 and 100 mg/kg bw, administered intraperitoneally 30 min prior to 9.5 Gy whole-body γ-irradiation was studied in Swiss albino mice. Radiation toxicity and mortality were observed during a period of 30 days and the percentage mortality was calculated. SM pretreatment with 50 mg/kg bw was found to be the most effective dose in maintaining body weight and in reducing the percentage mortality, while 100 mg/kg bw was found to be more effective in maintaining the spleen index and in stimulation of endogenous spleen colony-forming units. Pretreatment with SM (50 mg/kg bw) in mice irradiated with 15 Gy significantly reduced dead, inflammatory, mitotic and goblet cells in irradiated jejunum. SM at 50 mg/kg bw also increased crypt cells, maintained villus height, and prevented mucosal erosion. Nuclear enlargement in epithelial cells was found less in SM-treated mice compared with the irradiated control. The radiation-induced decrease in endogenous antioxidant enzymes (GSH, GST, catalase) and the increase in lipid peroxidation were also reduced by pretreatment with SM [50 and 100 mg/kg bw] at all monitored post-irradiation intervals. There was no protection at a dose less than 25 mg/kg bw.     

Effect of sesamol on radiation-induced cytotoxicity in Swiss albino mice

The radio-protective ability of sesamol (SM) at various doses viz., 0, 10, 25, 40, 50, 70 and 100mg/kg bw, administered intraperitoneally 30min prior to 9.5Gy whole-body gamma-irradiation was studied in Swiss albino mice. Radiation toxicity and mortality were observed during a period of 30 days and the percentage mortality was calculated. SM pretreatment with 50mg/kg bw was found to be the most effective dose in maintaining body weight and in reducing the percentage mortality, while 100mg/kg bw was found to be more effective in maintaining the spleen index and in stimulation of endogenous spleen colony-forming units. Pretreatment with SM (50mg/kg bw) in mice irradiated with 15Gy significantly reduced dead, inflammatory, mitotic and goblet cells in irradiated jejunum. SM at 50mg/kg bw also increased crypt cells, maintained villus height, and prevented mucosal erosion. Nuclear enlargement in epithelial cells was found less in SM-treated mice compared with the irradiated control. The radiation-induced decrease in endogenous antioxidant enzymes (GSH, GST, catalase) and the increase in lipid peroxidation were also reduced by pretreatment with SM [50 and 100mg/kg bw] at all monitored post-irradiation intervals. There was no protection at a dose less than 25mg/kg bw.     

Manganese pre-treatment attenuates cadmium induced hepatotoxicity in Swiss albino mice

Cadmium (Cd) is a soft, malleable bluish-white metal with low melting point, a ubiquitous heavy metal and an environmental pollutant, found in soil, water and air. The presence of Cd in the components of the environment such as air, soil and groundwater is to a large part due to human activity, and the general population is exposed mainly by contaminated drinking water or food. Manganese (Mn) is a component in many enzymes, which play an important role in counteracting oxidative stress. In vitro experiments have revealed the ability of Mn to scavenge oxygen free radicals generated in differently mediated lipid peroxidation (LPO) conditions. The aim of the present study was to investigate the in vivo preventive effect of Mn²⁺ pre-treatment on acute Cd-intoxication with regard to oxidative stress biomarker and antioxidant defense system in liver of Swiss albino mice. On exposure to Cd a significant increase in LPO levels, decrease in thiol content and induction in glutathione metabolizing enzyme were observed. Mn pre-treatment attenuated the modulation caused in the above-mentioned parameters due to acute Cd exposure in mice. In conclusion, the results from this study demonstrate that the protective effect of Mn in Cd-induced systemic toxicity in mice. Further investigations are required on the relation between Mn accumulation and resistance to oxidative stress and on the factors influencing Mn/Cd transport in rodents are needed to elucidate the molecular basis of this protective effect.     

Inhibition of adriamycin-induced micronuclei by desferrioxamine in Swiss albino mice

Swiss albino male mice, 6–8 weeks old, were treated i.p. with different doses of desferrioxamine dissolved in water for 7 days. Some of the mice in each group were injected i.p. with adriamycin (15 mg/kg) and killed after 30 or 24, 48 and 72 h. The femoral cells of mice in different groups were collected and studied. Desferrioxamine treatment failed to affect the incidence of micronuclei at doses of 125–250 mg/kg/day. Pretreatment with desferrioxamine was found to provide significant protection against adriamycin-induced micronuclei without interfering with its cytotoxic potential.     

Chronic unpredictable stress exacerbates 7,12-dimethylbenz (a) anthracene induced hepatotoxicity and nephrotoxicity in Swiss albino mice

Oxidative stress, a pervasive condition induced by stress has been implicated and recognized to be a prominent feature of various pathological states including cancer and their progression. The present study sought to validate the effectiveness of chronic unpredictable stress (CUS) on hepatic and renal toxicity in terms of alterations of various in vivo biochemical parameters, oxidative stress markers and the extent of DNA damage in Swiss albino mice. Animals were randomized into different groups based on their exposure to CUS alone, 7,12-dimethylbenz (a) anthracene (DMBA) alone (topical), DMBA-12-O-tetradecanoylphorbol-13-acetate (TPA) (topical), and exposure to CUS prior to DMBA or DMBA-TPA treatment, and sacrificed after 16 weeks of treatment. Prior exposure to CUS increased the pro-oxidant effect of carcinogen as depicted by significantly compromised levels of antioxidants; superoxide dismutase, catalase, glutathione-S-transferase, glutathione reductase, reduced glutathione in hepatic and renal tissues accompanied by a significant elevation of thiobarbituric acid reactive species (TBARS) as compared to DMBA alone or DMBA-TPA treatments. Loss of structural integrity at the cellular level due to stress-induced oxidative damage was demonstrated by significant increases in the hepatic levels of intracellular marker enzymes such as glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and alkaline phosphatase, and significantly reduced levels of uric acid in kidney tissues. The results of DNA damage studies further positively correlated with all the above biochemical measurements. Thus, exposure to physical or psychological stress may significantly enhance the hepatotoxic and nephrotoxic potential of carcinogens through enhanced oxidative stress even if the treatment is topical.     

Stimulatory effects of Cuminum cyminum and flavonoid glycoside on Cyclosporine-A and restraint stress induced immune-suppression in Swiss albino mice

Many herbs and spices are known to modulate the immune system and have been shown to restore the immunity in immuno-compromised individuals. Spices generally used to increase the taste and flavor of food also has the history of usage as an ayurvedic medicine. Therefore to explore the health modulating effects of Cuminum cyminum and to identify the active compound, immunomodulatory properties were evaluated using flowcytometry and ELISA in normal and immune-suppressed animals. C. cyminum and compound 1 stimulated the T cells and Th1 cytokines expression in normal animals. Swiss albino mice subjected to Cyclosporine-A induced immune-suppression were dosed orally with C. cyminum (25, 50, 100 and 200 mg/kg) on consecutive days. The results showed that administration significantly increased T cells (CD4 and CD8) count and Th1 predominant immune response in a dose dependent manner thereby suggesting immunomodulatory activity through modulation of T lymphocytes expression. In restraint stress induced immune-suppressed animals, compound 1 countered the depleted T lymphocytes, decreased the elevated corticosterone levels and size of adrenal glands and increased the weight of thymus and spleen. Based on the data we may conclude that C. cyminum is a potent immunomodulator and may develop as a lead to recover the immunity of immuno-compromised individuals.     

Occurrence of priapism after transient right MCAO in Swiss albino mice

Introduction: There are few reports about sexual problems in animal models after stroke. The aim of this paper is to report the occurrence of priapism after right MCAO in Swiss albino mice. In addition, we compared neurological score and apoptosis between the priapism-affected and unaffected mice.

Methods: Swiss albino mice were subjected to 45?min’ MCAO and 7?days’ reperfusion. Mice were observed before MCAO, then daily for 7?days to assess priapism. Neurological status and apoptosis (TUNEL assay) were assessed and compared in priapism and non-priapism mice.

Results: The results showed that the incidence of priapism after MCAO in Swiss albino mice were 65%. Priapism was detectable often at day 2 after stroke. Priapism-affected group had more severe behavioural deficits after stroke compared to non-priapism stroke mice.

Conclusion: Priapism after right MCAO is not rare in albino mice and could be considered as a marker of stroke severity. Further studies are needed to assess the incidence of priapism after stroke in other animal species used for stroke studies such as rat.