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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

The effect on cellular redox markers in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.Panel A. Effect on ROS-generation (intrecellular) was measured by fluorescence microscopy (DCF-DA) in Pb-exposed hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, SOD, CAT, GST, GPx, GSH and GR in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Values are expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). *Values differed significantly from pb-acetate control (p < 0.05). **Values differed significantly from pb-acetate control (p < 0.01). SOD unit, “U” is defined as μ-moles inhibition of NBT-reduction/min while CAT unit “U” is defined as H2O2 consumed/minute.
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pone.0139831.g002: The effect on cellular redox markers in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.Panel A. Effect on ROS-generation (intrecellular) was measured by fluorescence microscopy (DCF-DA) in Pb-exposed hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, SOD, CAT, GST, GPx, GSH and GR in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Values are expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). *Values differed significantly from pb-acetate control (p < 0.05). **Values differed significantly from pb-acetate control (p < 0.01). SOD unit, “U” is defined as μ-moles inhibition of NBT-reduction/min while CAT unit “U” is defined as H2O2 consumed/minute.

Mentions: Fig 2 represented the effect of AEIA against Pb-acetate induced alteration of ROS production, peroxidation of lipids, carbonylation of proteins and intracellular redox status in murine hepatocytes. The enhance production of intracellular ROS is a marker of augmented oxidative stress (Fig 2A). Generation of ROS (intracellular) was measured by microscopy (fluorescence) using fluorescent dye DCF. In this study, Pb-acetate exposure significantly increased intracellular ROS production in murine hepatocytes. The AEIA treatment could significantly attenuate the ROS production in hepatocytes. Lipid peroxidation is an indication of oxidative tissue damage caused by ROS. TBARS is a yard stick of lipid-peroxidation. In this study, Pb-acetate treatment significantly increased (p < 0.01) TBARS levels as compared to control hepatocytes (Fig 2B). However, AEIA treatment could significantly alleviate (p < 0.01) the degree of lipid-peroxidation in vitro evidenced by reduction of TBARS level. Pb-acetate intoxication caused a significant (p < 0.01) increase in the carbonylation of proteins in hepatocytes (Fig 2B). AEIA treatment could significantly reinstate (p < 0.05) the Pb-acetate mediated protein carbonylation in murine hepatocytes. Cellular antioxidant enzymes and GSH served in cellular defense mechanism during redox stress. In this study, the levels of CAT, SOD, GR, GST, GPx and GSH were reduced (p < 0.01) significantly in Pb-acetate treated hepatocytes (Fig 2B). However, AEIA treatment could significantly (p < 0.05–0.01) revert the levels of aforementioned redox markers to near normal status. The data set is available in S1 Table.


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)

The effect on cellular redox markers in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.Panel A. Effect on ROS-generation (intrecellular) was measured by fluorescence microscopy (DCF-DA) in Pb-exposed hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, SOD, CAT, GST, GPx, GSH and GR in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Values are expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). *Values differed significantly from pb-acetate control (p < 0.05). **Values differed significantly from pb-acetate control (p < 0.01). SOD unit, “U” is defined as μ-moles inhibition of NBT-reduction/min while CAT unit “U” is defined as H2O2 consumed/minute.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139831.g002: The effect on cellular redox markers in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.Panel A. Effect on ROS-generation (intrecellular) was measured by fluorescence microscopy (DCF-DA) in Pb-exposed hepatocytes in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, SOD, CAT, GST, GPx, GSH and GR in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA). Values are expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). *Values differed significantly from pb-acetate control (p < 0.05). **Values differed significantly from pb-acetate control (p < 0.01). SOD unit, “U” is defined as μ-moles inhibition of NBT-reduction/min while CAT unit “U” is defined as H2O2 consumed/minute.
Mentions: Fig 2 represented the effect of AEIA against Pb-acetate induced alteration of ROS production, peroxidation of lipids, carbonylation of proteins and intracellular redox status in murine hepatocytes. The enhance production of intracellular ROS is a marker of augmented oxidative stress (Fig 2A). Generation of ROS (intracellular) was measured by microscopy (fluorescence) using fluorescent dye DCF. In this study, Pb-acetate exposure significantly increased intracellular ROS production in murine hepatocytes. The AEIA treatment could significantly attenuate the ROS production in hepatocytes. Lipid peroxidation is an indication of oxidative tissue damage caused by ROS. TBARS is a yard stick of lipid-peroxidation. In this study, Pb-acetate treatment significantly increased (p < 0.01) TBARS levels as compared to control hepatocytes (Fig 2B). However, AEIA treatment could significantly alleviate (p < 0.01) the degree of lipid-peroxidation in vitro evidenced by reduction of TBARS level. Pb-acetate intoxication caused a significant (p < 0.01) increase in the carbonylation of proteins in hepatocytes (Fig 2B). AEIA treatment could significantly reinstate (p < 0.05) the Pb-acetate mediated protein carbonylation in murine hepatocytes. Cellular antioxidant enzymes and GSH served in cellular defense mechanism during redox stress. In this study, the levels of CAT, SOD, GR, GST, GPx and GSH were reduced (p < 0.01) significantly in Pb-acetate treated hepatocytes (Fig 2B). However, AEIA treatment could significantly (p < 0.05–0.01) revert the levels of aforementioned redox markers to near normal status. The data set is available in S1 Table.

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