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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 Bad, Bcl-2, cyt-C, caspase 9, caspase 3 and Apaf 1 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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Fig4: Respective western blot analysis of Bad, Bcl-2, cyt-C, caspase 9, caspase 3 and Apaf 1 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: Involvement of apoptosis indicates cellular damage under redox challenged cellular atmosphere. Apoptosis is synchronized by complex interactions between anti- and pro- apoptotic proteins followed by activation of caspases. In this study, the involvements of intrinsic and extrinsic pathways have been evaluated by western blot analysis. The immunoblot analysis of intrinsic transcription proteins have been depicted in Figure 4. The experimental observations suggested that NaAsO2 (10 μM) significantly up-regulated the pro-apoptotic (Bad) and down-regulated the anti-apoptotic (Bcl-2) proteins, which caused a significant (p < 0.01) increase in Bad/Bcl-2 ratio in isolated murine hepatocytes. Significantly increased (p < 0.01) expression of cytosolic Cyt C in association with activation (p < 0.01) of caspase 9 and 3 demonstrated the involvement of intrinsic apoptotic pathway in pathogenesis in hepatocytes incubated with NaAsO2 (10 μM). In present study, immunoblot analysis of Apaf 1 showed that NaAsO2 exposure caused a significant (p < 0.01) increase in the expression of Apaf 1. AEIA (400 μg/ml) co-treatment, however, could significantly inhibit NaAsO2-induced up-regulation of Cyt C (p < 0.01), caspase 9 (p < 0.05), caspase 3 (p < 0.05) and Apaf 1 (p < 0.05) along with reciprocate the regulation (p < 0.05) of Bad and Bcl-2.Figure 4


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 Bad, Bcl-2, cyt-C, caspase 9, caspase 3 and Apaf 1 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

Fig4: Respective western blot analysis of Bad, Bcl-2, cyt-C, caspase 9, caspase 3 and Apaf 1 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: Involvement of apoptosis indicates cellular damage under redox challenged cellular atmosphere. Apoptosis is synchronized by complex interactions between anti- and pro- apoptotic proteins followed by activation of caspases. In this study, the involvements of intrinsic and extrinsic pathways have been evaluated by western blot analysis. The immunoblot analysis of intrinsic transcription proteins have been depicted in Figure 4. The experimental observations suggested that NaAsO2 (10 μM) significantly up-regulated the pro-apoptotic (Bad) and down-regulated the anti-apoptotic (Bcl-2) proteins, which caused a significant (p < 0.01) increase in Bad/Bcl-2 ratio in isolated murine hepatocytes. Significantly increased (p < 0.01) expression of cytosolic Cyt C in association with activation (p < 0.01) of caspase 9 and 3 demonstrated the involvement of intrinsic apoptotic pathway in pathogenesis in hepatocytes incubated with NaAsO2 (10 μM). In present study, immunoblot analysis of Apaf 1 showed that NaAsO2 exposure caused a significant (p < 0.01) increase in the expression of Apaf 1. AEIA (400 μg/ml) co-treatment, however, could significantly inhibit NaAsO2-induced up-regulation of Cyt C (p < 0.01), caspase 9 (p < 0.05), caspase 3 (p < 0.05) and Apaf 1 (p < 0.05) along with reciprocate the regulation (p < 0.05) of Bad and Bcl-2.Figure 4

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