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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

Histological sections 100 x (A) and 400 x (B) of livers of experimental mice in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA).Untreated mice were kept as normal control to compare the structural changes caused by Pb-acetate. Yellow and red arrows represent normal portal vein and hepatocytes, respectively; dotted arrows represent the Pb-acetate mediated structural changes of portal vein (yellow), hepatocytes (red) with infiltrating leukocytes (blue) and lipid deposition (green). C. Hepatic lipid droplets accumulation was presented as the percentage of the blank area relative to the whole area of the photomicrograph (100 x, randomly selected area devoid of portal vein were selected). D. The incidence of inflammation was presented as the percentage of the inflamed hepatocytes region relative to the whole area of the photomicrograph (100 x, randomly selected area in devoid of portal vein were selected). E. The structural change of portal vein was represented as percentage of the blank area relative to the whole area of the photomicrograph (400 x, randomly selected areas containing one portal vein were selected). Values are expressed as mean ± SE, (n = 60). $ Values significantly differed from normal control (P < 0.05). # Values differ significantly from normal control (p < 0.01). **Values significantly differed from Pb-acetate control (P < 0.01).
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pone.0139831.g007: Histological sections 100 x (A) and 400 x (B) of livers of experimental mice in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA).Untreated mice were kept as normal control to compare the structural changes caused by Pb-acetate. Yellow and red arrows represent normal portal vein and hepatocytes, respectively; dotted arrows represent the Pb-acetate mediated structural changes of portal vein (yellow), hepatocytes (red) with infiltrating leukocytes (blue) and lipid deposition (green). C. Hepatic lipid droplets accumulation was presented as the percentage of the blank area relative to the whole area of the photomicrograph (100 x, randomly selected area devoid of portal vein were selected). D. The incidence of inflammation was presented as the percentage of the inflamed hepatocytes region relative to the whole area of the photomicrograph (100 x, randomly selected area in devoid of portal vein were selected). E. The structural change of portal vein was represented as percentage of the blank area relative to the whole area of the photomicrograph (400 x, randomly selected areas containing one portal vein were selected). Values are expressed as mean ± SE, (n = 60). $ Values significantly differed from normal control (P < 0.05). # Values differ significantly from normal control (p < 0.01). **Values significantly differed from Pb-acetate control (P < 0.01).

Mentions: The histological sections of livers of mice have been depicted in Fig 7A (x 100) and B (x 400). The liver section of Pb-acetate-intoxicated mice exhibited diffuse portal veins, inflamed hepatocytes with infiltrating leukocytes and lipid deposition when compared with the liver sections of normal mice. Histo-quantification revealed a significant elevation of % of lipid deposition (p < 0.01), % of the area of inflamed hepatocytes (p < 0.01) and the % of area of portal veins (p < 0.05) in Pb-intoxicated liver sections (Fig 7C–7E). AEIA treatment could significantly (p < 0.01) revert the lipid deposition and hepatic inflammation to near normal status, however, extract treatment could insignificantly (p < 0.05) arrest the dilation of portal vein. The histological sections of kidneys of mice have been shown in Fig 8A (x 100) and B (x 400). The kidney sections of Pb-acetate control mice exhibited thickening of bowman’s capsules and cellular damage with cloudy appearance of tubules when compared with normal control animals. Histo-quantification revealed that Pb-intoxication caused significant thickening in capsular space (p < 0.01) and cloudy swelling of renal tubules (p < 0.01) (Fig 8C and 8D). Treatment with AEIA significantly (p < 0.05–0.01) reduced the Pb-induced pathological changes and restores the histology near to normalcy. Histological sections of hearts have been revealed in Fig 9A (x 100) and 9B (x 400). Pb-intoxication caused significant damage of the interstitial tissues (p < 0.01) and muscle replacement by adipose tissues (p < 0.01) (Fig 9C and 9D). AEIA treatment could significantly (p < 0.01) arrest the damage in cardiac section. The histological sections of brains have been represented in Fig 10A (x 100) and 10B (x 400). Pb-intoxicated mice exhibited a significant increase in % of the vacuolated area of degenerated tissues (p < 0.01) and diffused edema (p < 0.01) as compared to normal control mice (Fig 10C and 10D). However, treatment with AEIA could significantly (p < 0.01) reinstate Pb-mediated toxic manifestation in brain. The sections of testes have been shown in Fig 11A (x 100) and 11B (x 400). The testes segment of Pb-acetate treated mice exhibited substantial disintegration of seminiferous tubules with loss of spermatogenic cells. The histoquantification of testicular sections were scored with Jhonsen score (Fig 11C). A significant (p < 0.01) reduction of Jhonsen score was observed in Pb-intoxicated mice. However, AEIA treatment could significantly (p < 0.01) reinstate the Jhonsen score and restore the histology near to normalcy.


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)

Histological sections 100 x (A) and 400 x (B) of livers of experimental mice in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA).Untreated mice were kept as normal control to compare the structural changes caused by Pb-acetate. Yellow and red arrows represent normal portal vein and hepatocytes, respectively; dotted arrows represent the Pb-acetate mediated structural changes of portal vein (yellow), hepatocytes (red) with infiltrating leukocytes (blue) and lipid deposition (green). C. Hepatic lipid droplets accumulation was presented as the percentage of the blank area relative to the whole area of the photomicrograph (100 x, randomly selected area devoid of portal vein were selected). D. The incidence of inflammation was presented as the percentage of the inflamed hepatocytes region relative to the whole area of the photomicrograph (100 x, randomly selected area in devoid of portal vein were selected). E. The structural change of portal vein was represented as percentage of the blank area relative to the whole area of the photomicrograph (400 x, randomly selected areas containing one portal vein were selected). Values are expressed as mean ± SE, (n = 60). $ Values significantly differed from normal control (P < 0.05). # Values differ significantly from normal control (p < 0.01). **Values significantly differed from Pb-acetate control (P < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139831.g007: Histological sections 100 x (A) and 400 x (B) of livers of experimental mice in the absence (Pb-acetate) and existence of AEIA (Pb-acetate + AEIA).Untreated mice were kept as normal control to compare the structural changes caused by Pb-acetate. Yellow and red arrows represent normal portal vein and hepatocytes, respectively; dotted arrows represent the Pb-acetate mediated structural changes of portal vein (yellow), hepatocytes (red) with infiltrating leukocytes (blue) and lipid deposition (green). C. Hepatic lipid droplets accumulation was presented as the percentage of the blank area relative to the whole area of the photomicrograph (100 x, randomly selected area devoid of portal vein were selected). D. The incidence of inflammation was presented as the percentage of the inflamed hepatocytes region relative to the whole area of the photomicrograph (100 x, randomly selected area in devoid of portal vein were selected). E. The structural change of portal vein was represented as percentage of the blank area relative to the whole area of the photomicrograph (400 x, randomly selected areas containing one portal vein were selected). Values are expressed as mean ± SE, (n = 60). $ Values significantly differed from normal control (P < 0.05). # Values differ significantly from normal control (p < 0.01). **Values significantly differed from Pb-acetate control (P < 0.01).
Mentions: The histological sections of livers of mice have been depicted in Fig 7A (x 100) and B (x 400). The liver section of Pb-acetate-intoxicated mice exhibited diffuse portal veins, inflamed hepatocytes with infiltrating leukocytes and lipid deposition when compared with the liver sections of normal mice. Histo-quantification revealed a significant elevation of % of lipid deposition (p < 0.01), % of the area of inflamed hepatocytes (p < 0.01) and the % of area of portal veins (p < 0.05) in Pb-intoxicated liver sections (Fig 7C–7E). AEIA treatment could significantly (p < 0.01) revert the lipid deposition and hepatic inflammation to near normal status, however, extract treatment could insignificantly (p < 0.05) arrest the dilation of portal vein. The histological sections of kidneys of mice have been shown in Fig 8A (x 100) and B (x 400). The kidney sections of Pb-acetate control mice exhibited thickening of bowman’s capsules and cellular damage with cloudy appearance of tubules when compared with normal control animals. Histo-quantification revealed that Pb-intoxication caused significant thickening in capsular space (p < 0.01) and cloudy swelling of renal tubules (p < 0.01) (Fig 8C and 8D). Treatment with AEIA significantly (p < 0.05–0.01) reduced the Pb-induced pathological changes and restores the histology near to normalcy. Histological sections of hearts have been revealed in Fig 9A (x 100) and 9B (x 400). Pb-intoxication caused significant damage of the interstitial tissues (p < 0.01) and muscle replacement by adipose tissues (p < 0.01) (Fig 9C and 9D). AEIA treatment could significantly (p < 0.01) arrest the damage in cardiac section. The histological sections of brains have been represented in Fig 10A (x 100) and 10B (x 400). Pb-intoxicated mice exhibited a significant increase in % of the vacuolated area of degenerated tissues (p < 0.01) and diffused edema (p < 0.01) as compared to normal control mice (Fig 10C and 10D). However, treatment with AEIA could significantly (p < 0.01) reinstate Pb-mediated toxic manifestation in brain. The sections of testes have been shown in Fig 11A (x 100) and 11B (x 400). The testes segment of Pb-acetate treated mice exhibited substantial disintegration of seminiferous tubules with loss of spermatogenic cells. The histoquantification of testicular sections were scored with Jhonsen score (Fig 11C). A significant (p < 0.01) reduction of Jhonsen score was observed in Pb-intoxicated mice. However, AEIA treatment could significantly (p < 0.01) reinstate the Jhonsen score and restore the histology near to normalcy.

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