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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

Respective western blot analysis of Fas, Bid and caspase 8 in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated 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 were expressed as mean ± SE (n = 3). #Values differed significantly (p < 0.01) from normal control. *Values differed significantly (p < 0.05) from NaAsO2 control. **Values differed significantly (p < 0.01) from NaAsO2 control.
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Fig5: Respective western blot analysis of Fas, Bid and caspase 8 in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated 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 were expressed as mean ± SE (n = 3). #Values differed significantly (p < 0.01) from normal control. *Values differed significantly (p < 0.05) from NaAsO2 control. **Values differed significantly (p < 0.01) from NaAsO2 control.

Mentions: To study the effect of mitochondria independent apoptosis (extrinsic pathway), immunoblot analysis of Bid, caspase-8 and FAS were performed (Figure 5). NaAsO2 (10 μM) caused significant (p < 0.01) up-regulation of Bid, FAS and caspase 8 in isolated murine hepatocytes, which indicated the involvement of extrinsic pathway of apoptosis, simultaneously. However, concurrent treatment of the cells with AEIA (400 μg/ml) could significantly (p < 0.05-0.01) prevent the As-mediated up-regulation of transcription levels of extrinsic apoptotic proteins.Figure 5


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)

Respective western blot analysis of Fas, Bid and caspase 8 in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated 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 were expressed as mean ± SE (n = 3). #Values differed significantly (p < 0.01) from normal control. *Values differed significantly (p < 0.05) from NaAsO2 control. **Values differed significantly (p < 0.01) from NaAsO2 control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig5: Respective western blot analysis of Fas, Bid and caspase 8 in absence (NaAsO2) and presence of AEIA (NaAsO2 + AEIA) in isolated 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 were expressed as mean ± SE (n = 3). #Values differed significantly (p < 0.01) from normal control. *Values differed significantly (p < 0.05) from NaAsO2 control. **Values differed significantly (p < 0.01) from NaAsO2 control.
Mentions: To study the effect of mitochondria independent apoptosis (extrinsic pathway), immunoblot analysis of Bid, caspase-8 and FAS were performed (Figure 5). NaAsO2 (10 μM) caused significant (p < 0.01) up-regulation of Bid, FAS and caspase 8 in isolated murine hepatocytes, which indicated the involvement of extrinsic pathway of apoptosis, simultaneously. However, concurrent treatment of the cells with AEIA (400 μg/ml) could significantly (p < 0.05-0.01) prevent the As-mediated up-regulation of transcription levels of extrinsic apoptotic proteins.Figure 5

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