Limits...
Water Spinach, Ipomoea aquatic (Convolvulaceae), Ameliorates Lead Toxicity by Inhibiting Oxidative Stress and Apoptosis.

Dewanjee S, Dua TK, Khanra R, Das S, Barma S, Joardar S, Bhattacharjee N, Zia-Ul-Haq M, Jaafar HZ - PLoS ONE (2015)

Bottom Line: The effects on the expressions of apoptotic signal proteins were estimated by western blotting.The extract may offer the protective effect via counteracting with Pb mediated oxidative stress and/or promoting the elimination of Pb by chelating.The presence of substantial quantities of flavonoids, phenolics and saponins would be responsible for the overall protective effect.

View Article: PubMed Central - PubMed

Affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India.

ABSTRACT

Background: Ipomoea aquatica (Convolvulaceae), an aquatic edible plant, is traditionally used against heavy metal toxicity in India. The current study intended to explore the protective role of edible (aqueous) extract of I. aquatica (AEIA) against experimentally induced Pb-intoxication.

Methods: The cytoprotective role of AEIA was measured on mouse hepatocytes by cell viability assay followed by Hoechst staining and flow cytometric assay. The effect on ROS production, lipid peroxidation, protein carbonylation, intracellular redox status were measured after incubating the hepatocytes with Pb-acetate (6.8 μM) along with AEIA (400 μg/ml). The effects on the expressions of apoptotic signal proteins were estimated by western blotting. The protective role of AEIA was measured by in vivo assay in mice. Haematological, serum biochemical, tissue redox status, Pb bioaccumulation and histological parameters were evaluated to estimate the protective role of AEIA (100 mg/kg) against Pb-acetate (5 mg/kg) intoxication.

Results: Pb-acetate treated hepatocytes showed a gradual reduction of cell viability dose-dependently with an IC50 value of 6.8 μM. Pb-acetate treated hepatocytes exhibited significantly enhanced levels (p < 0.01) of ROS production, lipid peroxidation, protein carbonylation with concomitant depletion (p < 0.01) of antioxidant enzymes and GSH. However, AEIA treatment could significantly restore the aforementioned parameters in murine hepatocytes near to normalcy. Besides, AEIA significantly reversed (p < 0.05-0.01) the alterations of transcription levels of apoptotic proteins viz. Bcl 2, Bad, Cyt C, Apaf-1, cleaved caspases [caspase 3, caspase 8 and caspase 9], Fas and Bid. In in vivo bioassay, Pb-acetate treatment caused significantly high intracellular Pb burden and oxidative pressure in the kidney, liver, heart, brain and testes in mice. In addition, the haematological and serum biochemical factors were changed significantly in Pb-acetate-treated animals. AEIA treatment restored significantly the evaluated-parameters to the near-normal position.

Conclusion: The extract may offer the protective effect via counteracting with Pb mediated oxidative stress and/or promoting the elimination of Pb by chelating. The presence of substantial quantities of flavonoids, phenolics and saponins would be responsible for the overall protective effect.

No MeSH data available.


Related in: MedlinePlus

Cell viability studies in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in vitro.Panel A. Effect of Pb-acetate at different concentrations in cell viability in mouse hepatocytes. Panel B. Concentration and time-dependent effect on cell viability studies in the absence (Pb-acetate) and existence of AEIA in isolated murine hepatocytes. Values are denoted as mean ± SE (n = 3). Panel C. Hoechst staining of murine hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel D. Percentage distribution of apoptotic and necrotic cells in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) by FCA.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4608788&req=5

pone.0139831.g001: Cell viability studies in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in vitro.Panel A. Effect of Pb-acetate at different concentrations in cell viability in mouse hepatocytes. Panel B. Concentration and time-dependent effect on cell viability studies in the absence (Pb-acetate) and existence of AEIA in isolated murine hepatocytes. Values are denoted as mean ± SE (n = 3). Panel C. Hoechst staining of murine hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel D. Percentage distribution of apoptotic and necrotic cells in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) by FCA.

Mentions: Cell viability indicates the degree of cytotoxity caused by any toxicant. Fig 1A depicted the cytotoxic effect of Pb-acetate on murine hepatocytes. Incubation of hepatocytes with Pb-acetate for 2 h exhibited a concentration dependent cytotoxic effect. The IC50 value was found to be ~ 6.8 μM. Based on observed IC50 value, Pb-acetate (6.8 μM) was used in the subsequent in vitro assays. The incubation of hepatocytes with AEIA (50–600 μg/ml) could significantly counteract with Pb-acetate induced cytotoxicity (Fig 1B). The extract could improve cell viability in a concentration dependent manner between 50–400 μg/ml, however, the cytoprotective effect was reduced beyond 400 μg/ml as observed at the dose of 600 μg/ml of AEIA. The reduction of cytoprotective effect beyond the dose of 400 μg/ml may be due to the prooxidative effects of AEIA at the concentration higher then optimal concentration require to exert physiological benefit. ‘Double-edged sword’ effect of exogenous antioxidants has been demonstrated in the literature [32]. Based on the observed effect AEIA (400 μg/ml) has been chosen for subsequent in vitro assays.


Water Spinach, Ipomoea aquatic (Convolvulaceae), Ameliorates Lead Toxicity by Inhibiting Oxidative Stress and Apoptosis.

Dewanjee S, Dua TK, Khanra R, Das S, Barma S, Joardar S, Bhattacharjee N, Zia-Ul-Haq M, Jaafar HZ - PLoS ONE (2015)

Cell viability studies in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in vitro.Panel A. Effect of Pb-acetate at different concentrations in cell viability in mouse hepatocytes. Panel B. Concentration and time-dependent effect on cell viability studies in the absence (Pb-acetate) and existence of AEIA in isolated murine hepatocytes. Values are denoted as mean ± SE (n = 3). Panel C. Hoechst staining of murine hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel D. Percentage distribution of apoptotic and necrotic cells in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) by FCA.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608788&req=5

pone.0139831.g001: Cell viability studies in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in vitro.Panel A. Effect of Pb-acetate at different concentrations in cell viability in mouse hepatocytes. Panel B. Concentration and time-dependent effect on cell viability studies in the absence (Pb-acetate) and existence of AEIA in isolated murine hepatocytes. Values are denoted as mean ± SE (n = 3). Panel C. Hoechst staining of murine hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel D. Percentage distribution of apoptotic and necrotic cells in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) by FCA.
Mentions: Cell viability indicates the degree of cytotoxity caused by any toxicant. Fig 1A depicted the cytotoxic effect of Pb-acetate on murine hepatocytes. Incubation of hepatocytes with Pb-acetate for 2 h exhibited a concentration dependent cytotoxic effect. The IC50 value was found to be ~ 6.8 μM. Based on observed IC50 value, Pb-acetate (6.8 μM) was used in the subsequent in vitro assays. The incubation of hepatocytes with AEIA (50–600 μg/ml) could significantly counteract with Pb-acetate induced cytotoxicity (Fig 1B). The extract could improve cell viability in a concentration dependent manner between 50–400 μg/ml, however, the cytoprotective effect was reduced beyond 400 μg/ml as observed at the dose of 600 μg/ml of AEIA. The reduction of cytoprotective effect beyond the dose of 400 μg/ml may be due to the prooxidative effects of AEIA at the concentration higher then optimal concentration require to exert physiological benefit. ‘Double-edged sword’ effect of exogenous antioxidants has been demonstrated in the literature [32]. Based on the observed effect AEIA (400 μg/ml) has been chosen for subsequent in vitro assays.

Bottom Line: The effects on the expressions of apoptotic signal proteins were estimated by western blotting.The extract may offer the protective effect via counteracting with Pb mediated oxidative stress and/or promoting the elimination of Pb by chelating.The presence of substantial quantities of flavonoids, phenolics and saponins would be responsible for the overall protective effect.

View Article: PubMed Central - PubMed

Affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India.

ABSTRACT

Background: Ipomoea aquatica (Convolvulaceae), an aquatic edible plant, is traditionally used against heavy metal toxicity in India. The current study intended to explore the protective role of edible (aqueous) extract of I. aquatica (AEIA) against experimentally induced Pb-intoxication.

Methods: The cytoprotective role of AEIA was measured on mouse hepatocytes by cell viability assay followed by Hoechst staining and flow cytometric assay. The effect on ROS production, lipid peroxidation, protein carbonylation, intracellular redox status were measured after incubating the hepatocytes with Pb-acetate (6.8 μM) along with AEIA (400 μg/ml). The effects on the expressions of apoptotic signal proteins were estimated by western blotting. The protective role of AEIA was measured by in vivo assay in mice. Haematological, serum biochemical, tissue redox status, Pb bioaccumulation and histological parameters were evaluated to estimate the protective role of AEIA (100 mg/kg) against Pb-acetate (5 mg/kg) intoxication.

Results: Pb-acetate treated hepatocytes showed a gradual reduction of cell viability dose-dependently with an IC50 value of 6.8 μM. Pb-acetate treated hepatocytes exhibited significantly enhanced levels (p < 0.01) of ROS production, lipid peroxidation, protein carbonylation with concomitant depletion (p < 0.01) of antioxidant enzymes and GSH. However, AEIA treatment could significantly restore the aforementioned parameters in murine hepatocytes near to normalcy. Besides, AEIA significantly reversed (p < 0.05-0.01) the alterations of transcription levels of apoptotic proteins viz. Bcl 2, Bad, Cyt C, Apaf-1, cleaved caspases [caspase 3, caspase 8 and caspase 9], Fas and Bid. In in vivo bioassay, Pb-acetate treatment caused significantly high intracellular Pb burden and oxidative pressure in the kidney, liver, heart, brain and testes in mice. In addition, the haematological and serum biochemical factors were changed significantly in Pb-acetate-treated animals. AEIA treatment restored significantly the evaluated-parameters to the near-normal position.

Conclusion: The extract may offer the protective effect via counteracting with Pb mediated oxidative stress and/or promoting the elimination of Pb by chelating. The presence of substantial quantities of flavonoids, phenolics and saponins would be responsible for the overall protective effect.

No MeSH data available.


Related in: MedlinePlus