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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

Respective western blot analysis of Fas, Bid and caspase 8 in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.The relative band intensities were measured and the normal control band was given an arbitrary value of 1. β-actin was used as a loading protein. Values are expressed as mean ± SE (n = 3). # Values differed (p < 0.01) significantly from normal control. *Values differed (p < 0.05) significantly from Pb-acetate control. **Values differed (p < 0.05) significantly from Pb-acetate control.
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pone.0139831.g004: Respective western blot analysis of Fas, Bid and caspase 8 in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.The relative band intensities were measured and the normal control band was given an arbitrary value of 1. β-actin was used as a loading protein. Values are expressed as mean ± SE (n = 3). # Values differed (p < 0.01) significantly from normal control. *Values differed (p < 0.05) significantly from Pb-acetate control. **Values differed (p < 0.05) significantly from Pb-acetate control.

Mentions: To study the effect of Pb-acetate in extrinsic factors’ mediated apoptosis, immunoblot analysis of FAS, Bid and cleaved caspase-8 were performed (Fig 4). Pb-acetate significantly (p < 0.01) up-regulated the FAS, Bid and caspase 8 in isolated murine hepatocytes, which indicated the involvement of extrinsic pathway of apoptosis, simultaneously.


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)

Respective western blot analysis of Fas, Bid and caspase 8 in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.The relative band intensities were measured and the normal control band was given an arbitrary value of 1. β-actin was used as a loading protein. Values are expressed as mean ± SE (n = 3). # Values differed (p < 0.01) significantly from normal control. *Values differed (p < 0.05) significantly from Pb-acetate control. **Values differed (p < 0.05) significantly from Pb-acetate control.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139831.g004: Respective western blot analysis of Fas, Bid and caspase 8 in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA) in murine hepatocytes.The relative band intensities were measured and the normal control band was given an arbitrary value of 1. β-actin was used as a loading protein. Values are expressed as mean ± SE (n = 3). # Values differed (p < 0.01) significantly from normal control. *Values differed (p < 0.05) significantly from Pb-acetate control. **Values differed (p < 0.05) significantly from Pb-acetate control.
Mentions: To study the effect of Pb-acetate in extrinsic factors’ mediated apoptosis, immunoblot analysis of FAS, Bid and cleaved caspase-8 were performed (Fig 4). Pb-acetate significantly (p < 0.01) up-regulated the FAS, Bid and caspase 8 in isolated murine hepatocytes, which indicated the involvement of extrinsic pathway of apoptosis, simultaneously.

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