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Ameliorative effect of water spinach, Ipomea aquatica (Convolvulaceae), against experimentally induced arsenic toxicity.

Dua TK, Dewanjee S, Gangopadhyay M, Khanra R, Zia-Ul-Haq M, De Feo V - J Transl Med (2015)

Bottom Line: In addition, the serum biochemical and haematological parameters were significantly (p < 0.05-0.01) altered in the NaAsO2-treated animals.However, concurrent administration of AEIA (100 mg/ml) could significantly reinstate the NaAsO2-induced pathogenesis.Presence of substantial quantities of dietary antioxidants within AEIA would be responsible for overall protective effect.

View Article: PubMed Central - PubMed

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

ABSTRACT

Background: Ipomea aquatica (Convolvulaceae) is traditionally used against Arsenic (As) poisoning in folk medicines in India. The present study was designed to explore the therapeutic role of aqueous extract of I. aquatica (AEIA) against As-intoxication.

Methods: AEIA was chemically standardized by spectroscopic and chromatographic analysis. The cytoprotective role of AEIA was measured on isolated murine hepatocytes. The effect on redox status were measured after incubating the hepatocytes with NaAsO2 (10 μM) + AEIA (400 μg/ml). The protective effect of AEIA (400 μg/ml) in expressions of apoptotic proteins were estimated in vitro. The protective role of AEIA was measured by in vivo assay in mice. Haematological, biochemical, As bioaccumulation and histological parameters were evaluated to ensure the protective role of AEIA (100 mg/kg) against NaAsO2 (10 mg/kg) intoxication.

Results: Phytochemical analysis revealed presence of substantial quantities of phenolics, flavonoids, saponins and ascorbic acid in AEIA. Incubation of murine hepatocytes with AEIA (0-400 μg/ml) + NaAsO2 (10 μM) exerted a concentration dependent cytoprotective effect. Incubation of murine hepatocytes with NaAsO2 (10 μM, ~ IC50) induced apoptosis via augmenting oxidative stress. NaAsO2 treated hepatocytes exhibited significantly (p < 0.01) enhanced levels of ROS production, lipid peroxidation and protein carbonylation with concomitant depletion of antioxidant enzymes (p < 0.05-0.01) and GSH (p < 0.01) levels. However, AEIA (400 μg/ml) + NaAsO2 (10 μM) could significantly (p < 0.05-0.01) reinstate the aforementioned parameters to near-normal status. Besides, AEIA (400 μg/ml) could significantly counteract (p <0.05-0.01) ROS mediated alteration in the expressions of apoptotic proteins viz. Bcl-2, BAD, Cyt C, Apaf 1, caspases, Fas and Bid. In in vivo bioassay, NaAsO2 (10 mg/kg) treatment in mice caused significantly (p < 0.05-0.01) elevated As bioaccumulation, ATP levels, DNA fragmentations and oxidative stress in the liver, kidney, heart, brain and testes along with alteration in cytoarchitecture of these organs. In addition, the serum biochemical and haematological parameters were significantly (p < 0.05-0.01) altered in the NaAsO2-treated animals. However, concurrent administration of AEIA (100 mg/ml) could significantly reinstate the NaAsO2-induced pathogenesis.

Conclusion: Presence of substantial quantities of dietary antioxidants within AEIA would be responsible for overall protective effect.

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Related in: MedlinePlus

The effect on cellular redox markers in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Panel A. Effect on intracellular ROS production was detected by fluorescence microscopy using DCF-DA in Cd-exposed hepatocytes in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, CAT, SOD, GR, GPx, GST and GSH in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Values were expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). $ Values differ significantly from normal control (p < 0.05). *Values differed significantly from NaAsO2 control (p < 0.05). **Values differed significantly from NaAsO2 control (p < 0.01). CAT unit, ‘U’ is defined as μmoles of H2O2 consumed per minute. SOD unit, ‘U’ is defined as the μmoles inhibition of NBT reduction per minute.
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Fig3: The effect on cellular redox markers in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Panel A. Effect on intracellular ROS production was detected by fluorescence microscopy using DCF-DA in Cd-exposed hepatocytes in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, CAT, SOD, GR, GPx, GST and GSH in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Values were expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). $ Values differ significantly from normal control (p < 0.05). *Values differed significantly from NaAsO2 control (p < 0.05). **Values differed significantly from NaAsO2 control (p < 0.01). CAT unit, ‘U’ is defined as μmoles of H2O2 consumed per minute. SOD unit, ‘U’ is defined as the μmoles inhibition of NBT reduction per minute.

Mentions: Figure 3 represented the effect of AEIA against NaAsO2 induced alteration of antioxidant markers in isolated in murine hepatocytes. Intracellular ROS production plays a principle role in NaAsO2 induced oxidative stress. Production of intracellular ROS was measured by fluorescence microscopy using fluorescent dye DCF (Figure 3A). It has been observed that NaAsO2 (10 μM) exposure led to an increased production of intracellular ROS, which could be prevented by simultaneous incubation of NaAsO2 (10 μM) along with AEIA (400 μg/ml). The extents of lipid peroxidation (TBARS level) and protein carbonylation were significantly (p < 0.01) increased in the hepatocytes incubated with NaAsO2 (10 μM) when compared with control hepatocytes (Figure 3B). Simultaneous incubation of AEIA (400 μg/ml) and NaAsO2 (10 μM) could significantly inhibit lipid peroxidation (p < 0.05) and protein carbonylation (p < 0.01) when compared to NaAsO2 (10 μM) treated group. Cellular antioxidant enzymes protect biological macromolecules from oxidative damage. In this study, the levels of CAT, SOD, GPx, GR and GST were significantly (p < 0.05-0.01) reduced in NaAsO2 treated hepatocytes (Figure 3B). However, concurrent incubation of hepatocytes with AEIA (400 μg/ml) and NaAsO2 (10 μM) could significantly (p < 0.05-0.01) revert the levels of antioxidant enzymes near to normalcy.Figure 3


Ameliorative effect of water spinach, Ipomea aquatica (Convolvulaceae), against experimentally induced arsenic toxicity.

Dua TK, Dewanjee S, Gangopadhyay M, Khanra R, Zia-Ul-Haq M, De Feo V - J Transl Med (2015)

The effect on cellular redox markers in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Panel A. Effect on intracellular ROS production was detected by fluorescence microscopy using DCF-DA in Cd-exposed hepatocytes in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, CAT, SOD, GR, GPx, GST and GSH in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Values were expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). $ Values differ significantly from normal control (p < 0.05). *Values differed significantly from NaAsO2 control (p < 0.05). **Values differed significantly from NaAsO2 control (p < 0.01). CAT unit, ‘U’ is defined as μmoles of H2O2 consumed per minute. SOD unit, ‘U’ is defined as the μmoles inhibition of NBT reduction per minute.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359489&req=5

Fig3: The effect on cellular redox markers in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Panel A. Effect on intracellular ROS production was detected by fluorescence microscopy using DCF-DA in Cd-exposed hepatocytes in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA). Panel B. Effect on antioxidant parameters viz. lipid peroxidation, protein carbonylation, CAT, SOD, GR, GPx, GST and GSH in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated murine hepatocytes. Values were expressed as mean ± SE (n = 3). #Values differed significantly from normal control (p < 0.01). $ Values differ significantly from normal control (p < 0.05). *Values differed significantly from NaAsO2 control (p < 0.05). **Values differed significantly from NaAsO2 control (p < 0.01). CAT unit, ‘U’ is defined as μmoles of H2O2 consumed per minute. SOD unit, ‘U’ is defined as the μmoles inhibition of NBT reduction per minute.
Mentions: Figure 3 represented the effect of AEIA against NaAsO2 induced alteration of antioxidant markers in isolated in murine hepatocytes. Intracellular ROS production plays a principle role in NaAsO2 induced oxidative stress. Production of intracellular ROS was measured by fluorescence microscopy using fluorescent dye DCF (Figure 3A). It has been observed that NaAsO2 (10 μM) exposure led to an increased production of intracellular ROS, which could be prevented by simultaneous incubation of NaAsO2 (10 μM) along with AEIA (400 μg/ml). The extents of lipid peroxidation (TBARS level) and protein carbonylation were significantly (p < 0.01) increased in the hepatocytes incubated with NaAsO2 (10 μM) when compared with control hepatocytes (Figure 3B). Simultaneous incubation of AEIA (400 μg/ml) and NaAsO2 (10 μM) could significantly inhibit lipid peroxidation (p < 0.05) and protein carbonylation (p < 0.01) when compared to NaAsO2 (10 μM) treated group. Cellular antioxidant enzymes protect biological macromolecules from oxidative damage. In this study, the levels of CAT, SOD, GPx, GR and GST were significantly (p < 0.05-0.01) reduced in NaAsO2 treated hepatocytes (Figure 3B). However, concurrent incubation of hepatocytes with AEIA (400 μg/ml) and NaAsO2 (10 μM) could significantly (p < 0.05-0.01) revert the levels of antioxidant enzymes near to normalcy.Figure 3

Bottom Line: In addition, the serum biochemical and haematological parameters were significantly (p < 0.05-0.01) altered in the NaAsO2-treated animals.However, concurrent administration of AEIA (100 mg/ml) could significantly reinstate the NaAsO2-induced pathogenesis.Presence of substantial quantities of dietary antioxidants within AEIA would be responsible for overall protective effect.

View Article: PubMed Central - PubMed

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

ABSTRACT

Background: Ipomea aquatica (Convolvulaceae) is traditionally used against Arsenic (As) poisoning in folk medicines in India. The present study was designed to explore the therapeutic role of aqueous extract of I. aquatica (AEIA) against As-intoxication.

Methods: AEIA was chemically standardized by spectroscopic and chromatographic analysis. The cytoprotective role of AEIA was measured on isolated murine hepatocytes. The effect on redox status were measured after incubating the hepatocytes with NaAsO2 (10 μM) + AEIA (400 μg/ml). The protective effect of AEIA (400 μg/ml) in expressions of apoptotic proteins were estimated in vitro. The protective role of AEIA was measured by in vivo assay in mice. Haematological, biochemical, As bioaccumulation and histological parameters were evaluated to ensure the protective role of AEIA (100 mg/kg) against NaAsO2 (10 mg/kg) intoxication.

Results: Phytochemical analysis revealed presence of substantial quantities of phenolics, flavonoids, saponins and ascorbic acid in AEIA. Incubation of murine hepatocytes with AEIA (0-400 μg/ml) + NaAsO2 (10 μM) exerted a concentration dependent cytoprotective effect. Incubation of murine hepatocytes with NaAsO2 (10 μM, ~ IC50) induced apoptosis via augmenting oxidative stress. NaAsO2 treated hepatocytes exhibited significantly (p < 0.01) enhanced levels of ROS production, lipid peroxidation and protein carbonylation with concomitant depletion of antioxidant enzymes (p < 0.05-0.01) and GSH (p < 0.01) levels. However, AEIA (400 μg/ml) + NaAsO2 (10 μM) could significantly (p < 0.05-0.01) reinstate the aforementioned parameters to near-normal status. Besides, AEIA (400 μg/ml) could significantly counteract (p <0.05-0.01) ROS mediated alteration in the expressions of apoptotic proteins viz. Bcl-2, BAD, Cyt C, Apaf 1, caspases, Fas and Bid. In in vivo bioassay, NaAsO2 (10 mg/kg) treatment in mice caused significantly (p < 0.05-0.01) elevated As bioaccumulation, ATP levels, DNA fragmentations and oxidative stress in the liver, kidney, heart, brain and testes along with alteration in cytoarchitecture of these organs. In addition, the serum biochemical and haematological parameters were significantly (p < 0.05-0.01) altered in the NaAsO2-treated animals. However, concurrent administration of AEIA (100 mg/ml) could significantly reinstate the NaAsO2-induced pathogenesis.

Conclusion: Presence of substantial quantities of dietary antioxidants within AEIA would be responsible for overall protective effect.

Show MeSH
Related in: MedlinePlus