DNA stability and lipid peroxidation in vitamin E-deficient rats in vivo and colon cells in vitro

Modulation by the dietary anthocyanin, cyanidin-3-glycoside

Susan J Duthie, Peter T Gardner, Philip C Morrice, Sharon Wood, Lynn P Pirie, Charles Bestwick, Lesley Milne, Garry G Duthie

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background Fruit and vegetable consumption protects against cancer. This is attributed in part to antioxidants such as vitamin E combating oxidative DNA damage. Anthocyanins are found in significant concentrations in the human diet. However, it remains to be established whether they are bioactive in vivo. Aim To investigate the consequence both of vitamin E deficiency on oxidative damage to DNA and lipids and the cytoprotective effect of nutritionally relevant levels of cyanidin-3-glycoside both in vivo in rats and in vitro in human colonocytes. Methods Male Rowett Hooded Lister rats were fed a diet containing less than 0.5 mg/kg vitamin E or a vitamin E supplemented control diet containing 100 mg dα-tocopherol acetate/kg. Half of the controls and vitamin E-deficient rats received cyanidin-3-glycoside ( 100 mg/kg). After 12 weeks endogenous DNA stability in rat lymphocytes ( strand breaks and oxidised bases) and response to oxidative stress ex vivo ( H2O2; 200 μ M) was measured by single cell gel electrophoresis (SCGE). Tissue levels of 8-oxo-7,8-dihydro-2'- deoxyguanosine (8-Oxo-dG) were measured by HPLC with EC detection. Dα-tocopherol and lipid peroxidation products ( thiobarbituric acid reactive substances; TBARS) were measured by HPLC. Rat plasma pyruvate kinase and the production of reactive oxygen by phagocytes were detected spectrophotometrically and by flow cytometry respectively. Immortalised human colon epithelial cells ( HCEC) were preincubated in vitro with the anthocyanins cyanidin and cyanidin-3-glycoside and the flavonol quercetin ( all 50 μ M) before exposure to H2O2 ( 200 μ M). DNA damage was measured by SCGE as above. Results Plasma and liver dα-tocopherol declined progressively over 12 weeks in rats made vitamin E deficient. Lipid peroxidation was increased significantly in plasma, liver and red cells. Reactive oxygen levels in phagocytes and plasma pyruvate kinase were increased. Vitamin E deficiency did not affect DNA stability in rat lymphocytes, liver or colon. Cyanidin-3-glycoside did not alter lipid peroxidation or DNA damage in rats. However, it was chemoprotective against DNA damage in human colonocytes. DNA strand breakage was decreased 38.8 +/- 2.2 % after pretreatment with anthocyanin. Conclusion while it is accepted that vitamin E alters lipid oxidation in vivo, its role in maintaining DNA stability remains unclear. Moreover, whereas cyanidin-3- glycoside protects against oxidative DNA damage in vitro, at nutritionally relevant concentrations it is ineffective against oxidative stress in vivo.

Original languageEnglish
Pages (from-to)195-203
Number of pages9
JournalEuropean Journal of Nutrition
Volume44
Issue number4
DOIs
Publication statusPublished - Jun 2005

Keywords

  • cyanidin-3-glycoside
  • vitamin E deficiency
  • DNA stability
  • lipid peroxides
  • rat
  • tert-butyl-hydroperoxide
  • oxidative damage
  • uracil misincorporation
  • cyanidin 3-o-beta-d-glucoside
  • model systems
  • antioxidants
  • tocopherol
  • flavonoids
  • extract
  • plasma
  • cyanidin–3–glycoside
  • vitamin E deficiency
  • DNA stability
  • lipid peroxides

Cite this

@article{df0585b9aae349d0a9d05e8a9dbc6b88,
title = "DNA stability and lipid peroxidation in vitamin E-deficient rats in vivo and colon cells in vitro: Modulation by the dietary anthocyanin, cyanidin-3-glycoside",
abstract = "Background Fruit and vegetable consumption protects against cancer. This is attributed in part to antioxidants such as vitamin E combating oxidative DNA damage. Anthocyanins are found in significant concentrations in the human diet. However, it remains to be established whether they are bioactive in vivo. Aim To investigate the consequence both of vitamin E deficiency on oxidative damage to DNA and lipids and the cytoprotective effect of nutritionally relevant levels of cyanidin-3-glycoside both in vivo in rats and in vitro in human colonocytes. Methods Male Rowett Hooded Lister rats were fed a diet containing less than 0.5 mg/kg vitamin E or a vitamin E supplemented control diet containing 100 mg dα-tocopherol acetate/kg. Half of the controls and vitamin E-deficient rats received cyanidin-3-glycoside ( 100 mg/kg). After 12 weeks endogenous DNA stability in rat lymphocytes ( strand breaks and oxidised bases) and response to oxidative stress ex vivo ( H2O2; 200 μ M) was measured by single cell gel electrophoresis (SCGE). Tissue levels of 8-oxo-7,8-dihydro-2'- deoxyguanosine (8-Oxo-dG) were measured by HPLC with EC detection. Dα-tocopherol and lipid peroxidation products ( thiobarbituric acid reactive substances; TBARS) were measured by HPLC. Rat plasma pyruvate kinase and the production of reactive oxygen by phagocytes were detected spectrophotometrically and by flow cytometry respectively. Immortalised human colon epithelial cells ( HCEC) were preincubated in vitro with the anthocyanins cyanidin and cyanidin-3-glycoside and the flavonol quercetin ( all 50 μ M) before exposure to H2O2 ( 200 μ M). DNA damage was measured by SCGE as above. Results Plasma and liver dα-tocopherol declined progressively over 12 weeks in rats made vitamin E deficient. Lipid peroxidation was increased significantly in plasma, liver and red cells. Reactive oxygen levels in phagocytes and plasma pyruvate kinase were increased. Vitamin E deficiency did not affect DNA stability in rat lymphocytes, liver or colon. Cyanidin-3-glycoside did not alter lipid peroxidation or DNA damage in rats. However, it was chemoprotective against DNA damage in human colonocytes. DNA strand breakage was decreased 38.8 +/- 2.2 {\%} after pretreatment with anthocyanin. Conclusion while it is accepted that vitamin E alters lipid oxidation in vivo, its role in maintaining DNA stability remains unclear. Moreover, whereas cyanidin-3- glycoside protects against oxidative DNA damage in vitro, at nutritionally relevant concentrations it is ineffective against oxidative stress in vivo.",
keywords = "cyanidin-3-glycoside, vitamin E deficiency, DNA stability, lipid peroxides, rat, tert-butyl-hydroperoxide, oxidative damage, uracil misincorporation, cyanidin 3-o-beta-d-glucoside, model systems, antioxidants, tocopherol, flavonoids, extract, plasma, cyanidin–3–glycoside , vitamin E deficiency , DNA stability , lipid peroxides",
author = "Duthie, {Susan J} and Gardner, {Peter T} and Morrice, {Philip C} and Sharon Wood and Pirie, {Lynn P} and Charles Bestwick and Lesley Milne and Duthie, {Garry G}",
year = "2005",
month = "6",
doi = "10.1007/s00394-004-0511-1",
language = "English",
volume = "44",
pages = "195--203",
journal = "European Journal of Nutrition",
issn = "1436-6207",
publisher = "D. Steinkopff-Verlag",
number = "4",

}

TY - JOUR

T1 - DNA stability and lipid peroxidation in vitamin E-deficient rats in vivo and colon cells in vitro

T2 - Modulation by the dietary anthocyanin, cyanidin-3-glycoside

AU - Duthie, Susan J

AU - Gardner, Peter T

AU - Morrice, Philip C

AU - Wood, Sharon

AU - Pirie, Lynn P

AU - Bestwick, Charles

AU - Milne, Lesley

AU - Duthie, Garry G

PY - 2005/6

Y1 - 2005/6

N2 - Background Fruit and vegetable consumption protects against cancer. This is attributed in part to antioxidants such as vitamin E combating oxidative DNA damage. Anthocyanins are found in significant concentrations in the human diet. However, it remains to be established whether they are bioactive in vivo. Aim To investigate the consequence both of vitamin E deficiency on oxidative damage to DNA and lipids and the cytoprotective effect of nutritionally relevant levels of cyanidin-3-glycoside both in vivo in rats and in vitro in human colonocytes. Methods Male Rowett Hooded Lister rats were fed a diet containing less than 0.5 mg/kg vitamin E or a vitamin E supplemented control diet containing 100 mg dα-tocopherol acetate/kg. Half of the controls and vitamin E-deficient rats received cyanidin-3-glycoside ( 100 mg/kg). After 12 weeks endogenous DNA stability in rat lymphocytes ( strand breaks and oxidised bases) and response to oxidative stress ex vivo ( H2O2; 200 μ M) was measured by single cell gel electrophoresis (SCGE). Tissue levels of 8-oxo-7,8-dihydro-2'- deoxyguanosine (8-Oxo-dG) were measured by HPLC with EC detection. Dα-tocopherol and lipid peroxidation products ( thiobarbituric acid reactive substances; TBARS) were measured by HPLC. Rat plasma pyruvate kinase and the production of reactive oxygen by phagocytes were detected spectrophotometrically and by flow cytometry respectively. Immortalised human colon epithelial cells ( HCEC) were preincubated in vitro with the anthocyanins cyanidin and cyanidin-3-glycoside and the flavonol quercetin ( all 50 μ M) before exposure to H2O2 ( 200 μ M). DNA damage was measured by SCGE as above. Results Plasma and liver dα-tocopherol declined progressively over 12 weeks in rats made vitamin E deficient. Lipid peroxidation was increased significantly in plasma, liver and red cells. Reactive oxygen levels in phagocytes and plasma pyruvate kinase were increased. Vitamin E deficiency did not affect DNA stability in rat lymphocytes, liver or colon. Cyanidin-3-glycoside did not alter lipid peroxidation or DNA damage in rats. However, it was chemoprotective against DNA damage in human colonocytes. DNA strand breakage was decreased 38.8 +/- 2.2 % after pretreatment with anthocyanin. Conclusion while it is accepted that vitamin E alters lipid oxidation in vivo, its role in maintaining DNA stability remains unclear. Moreover, whereas cyanidin-3- glycoside protects against oxidative DNA damage in vitro, at nutritionally relevant concentrations it is ineffective against oxidative stress in vivo.

AB - Background Fruit and vegetable consumption protects against cancer. This is attributed in part to antioxidants such as vitamin E combating oxidative DNA damage. Anthocyanins are found in significant concentrations in the human diet. However, it remains to be established whether they are bioactive in vivo. Aim To investigate the consequence both of vitamin E deficiency on oxidative damage to DNA and lipids and the cytoprotective effect of nutritionally relevant levels of cyanidin-3-glycoside both in vivo in rats and in vitro in human colonocytes. Methods Male Rowett Hooded Lister rats were fed a diet containing less than 0.5 mg/kg vitamin E or a vitamin E supplemented control diet containing 100 mg dα-tocopherol acetate/kg. Half of the controls and vitamin E-deficient rats received cyanidin-3-glycoside ( 100 mg/kg). After 12 weeks endogenous DNA stability in rat lymphocytes ( strand breaks and oxidised bases) and response to oxidative stress ex vivo ( H2O2; 200 μ M) was measured by single cell gel electrophoresis (SCGE). Tissue levels of 8-oxo-7,8-dihydro-2'- deoxyguanosine (8-Oxo-dG) were measured by HPLC with EC detection. Dα-tocopherol and lipid peroxidation products ( thiobarbituric acid reactive substances; TBARS) were measured by HPLC. Rat plasma pyruvate kinase and the production of reactive oxygen by phagocytes were detected spectrophotometrically and by flow cytometry respectively. Immortalised human colon epithelial cells ( HCEC) were preincubated in vitro with the anthocyanins cyanidin and cyanidin-3-glycoside and the flavonol quercetin ( all 50 μ M) before exposure to H2O2 ( 200 μ M). DNA damage was measured by SCGE as above. Results Plasma and liver dα-tocopherol declined progressively over 12 weeks in rats made vitamin E deficient. Lipid peroxidation was increased significantly in plasma, liver and red cells. Reactive oxygen levels in phagocytes and plasma pyruvate kinase were increased. Vitamin E deficiency did not affect DNA stability in rat lymphocytes, liver or colon. Cyanidin-3-glycoside did not alter lipid peroxidation or DNA damage in rats. However, it was chemoprotective against DNA damage in human colonocytes. DNA strand breakage was decreased 38.8 +/- 2.2 % after pretreatment with anthocyanin. Conclusion while it is accepted that vitamin E alters lipid oxidation in vivo, its role in maintaining DNA stability remains unclear. Moreover, whereas cyanidin-3- glycoside protects against oxidative DNA damage in vitro, at nutritionally relevant concentrations it is ineffective against oxidative stress in vivo.

KW - cyanidin-3-glycoside

KW - vitamin E deficiency

KW - DNA stability

KW - lipid peroxides

KW - rat

KW - tert-butyl-hydroperoxide

KW - oxidative damage

KW - uracil misincorporation

KW - cyanidin 3-o-beta-d-glucoside

KW - model systems

KW - antioxidants

KW - tocopherol

KW - flavonoids

KW - extract

KW - plasma

KW - cyanidin–3–glycoside

KW - vitamin E deficiency

KW - DNA stability

KW - lipid peroxides

U2 - 10.1007/s00394-004-0511-1

DO - 10.1007/s00394-004-0511-1

M3 - Article

VL - 44

SP - 195

EP - 203

JO - European Journal of Nutrition

JF - European Journal of Nutrition

SN - 1436-6207

IS - 4

ER -