Effects of beta-carotene on antioxidant enzyme activity, intracellular reactive oxygen and membrane integrity within post confluent Caco-2 intestinal cells

Charles Bestwick, Lesley Milne

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

As encountered with a plethora of other natural products, the antioxidant activity of beta-carotene has been proposed as one of the mechanisms by which diets rich in this pro-vitamin A active carotenoid apparently afford chemoprevention. Here, we report the ability of beta-carotene to alter endogenous reactive oxygen levels and antioxidant defences within non-stressed 'differentiated' monolayers of an intestinal epithelial cell line (Caco-2) and to subsequently effect resistance to general oxidative insult. The differentiated monolayers efficiently absorbed beta-carotene. Between 3 and 8 days post confluence, cultures exhibited a progressive increase in antioxidant enzyme activity and a corresponding reduction to intracellular ROS levels. The profile for antioxidant enzyme activity was unaffected by sustained daily supplementation with beta-carotene. However, after two daily treatments with 50 mu M beta-carotene intracellular ROS levels were significantly reduced and there was a trend towards reduced intracellular ROS within monolayers subject to five daily treatments with 0.5 and 5 mu M beta-carotene. Prolonged supplementation with 0.1 and 0.5 mu M beta-carotene or short supplementation periods with 5 and 50 mu M beta-carotene did not alter susceptibility to H2O2. However, cultures treated daily between 3 and 8 days post confluence with 5 or 50 mu M beta-carotene exhibited enhanced LDH release, increased non-adherence and enhanced Trypan blue staining when challenged with 10 mM H2O2. In the absence of H2O2, the beta-carotene treatments were not overtly toxic to the monolayers. These results indicate that beta-carotene does not enhance antioxidant defences within Caco-2 monolayers. The enhancement of H2O2 toxicity by persistent, high doses of beta-carotene may contribute to the failure of this carotenoid to protect high risk individuals from certain degenerative conditions. (C) 1999 Elsevier Science B.V. All rights reserved.

Original languageEnglish
Pages (from-to)47-55
Number of pages9
JournalBiochimica et Biophysica Acta (BBA) - General Subjects
Volume1474
Issue number1
Early online date25 Feb 2000
DOIs
Publication statusPublished - 6 Mar 2000

Keywords

  • beta-carotene
  • antioxidant
  • reactive oxygen species
  • membrane damage
  • cell culture
  • Caco-2
  • free-radicals
  • lipid-peroxidation
  • cancer prevention
  • vitamin-E
  • supplememtation
  • vegetables
  • chemoprevention
  • catalase
  • diseases
  • systems

Cite this

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title = "Effects of beta-carotene on antioxidant enzyme activity, intracellular reactive oxygen and membrane integrity within post confluent Caco-2 intestinal cells",
abstract = "As encountered with a plethora of other natural products, the antioxidant activity of beta-carotene has been proposed as one of the mechanisms by which diets rich in this pro-vitamin A active carotenoid apparently afford chemoprevention. Here, we report the ability of beta-carotene to alter endogenous reactive oxygen levels and antioxidant defences within non-stressed 'differentiated' monolayers of an intestinal epithelial cell line (Caco-2) and to subsequently effect resistance to general oxidative insult. The differentiated monolayers efficiently absorbed beta-carotene. Between 3 and 8 days post confluence, cultures exhibited a progressive increase in antioxidant enzyme activity and a corresponding reduction to intracellular ROS levels. The profile for antioxidant enzyme activity was unaffected by sustained daily supplementation with beta-carotene. However, after two daily treatments with 50 mu M beta-carotene intracellular ROS levels were significantly reduced and there was a trend towards reduced intracellular ROS within monolayers subject to five daily treatments with 0.5 and 5 mu M beta-carotene. Prolonged supplementation with 0.1 and 0.5 mu M beta-carotene or short supplementation periods with 5 and 50 mu M beta-carotene did not alter susceptibility to H2O2. However, cultures treated daily between 3 and 8 days post confluence with 5 or 50 mu M beta-carotene exhibited enhanced LDH release, increased non-adherence and enhanced Trypan blue staining when challenged with 10 mM H2O2. In the absence of H2O2, the beta-carotene treatments were not overtly toxic to the monolayers. These results indicate that beta-carotene does not enhance antioxidant defences within Caco-2 monolayers. The enhancement of H2O2 toxicity by persistent, high doses of beta-carotene may contribute to the failure of this carotenoid to protect high risk individuals from certain degenerative conditions. (C) 1999 Elsevier Science B.V. All rights reserved.",
keywords = "beta-carotene, antioxidant, reactive oxygen species, membrane damage, cell culture, Caco-2, free-radicals, lipid-peroxidation, cancer prevention, vitamin-E, supplememtation, vegetables, chemoprevention, catalase, diseases, systems",
author = "Charles Bestwick and Lesley Milne",
year = "2000",
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doi = "10.1016/S0304-4165(99)00212-3 |",
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pages = "47--55",
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TY - JOUR

T1 - Effects of beta-carotene on antioxidant enzyme activity, intracellular reactive oxygen and membrane integrity within post confluent Caco-2 intestinal cells

AU - Bestwick, Charles

AU - Milne, Lesley

PY - 2000/3/6

Y1 - 2000/3/6

N2 - As encountered with a plethora of other natural products, the antioxidant activity of beta-carotene has been proposed as one of the mechanisms by which diets rich in this pro-vitamin A active carotenoid apparently afford chemoprevention. Here, we report the ability of beta-carotene to alter endogenous reactive oxygen levels and antioxidant defences within non-stressed 'differentiated' monolayers of an intestinal epithelial cell line (Caco-2) and to subsequently effect resistance to general oxidative insult. The differentiated monolayers efficiently absorbed beta-carotene. Between 3 and 8 days post confluence, cultures exhibited a progressive increase in antioxidant enzyme activity and a corresponding reduction to intracellular ROS levels. The profile for antioxidant enzyme activity was unaffected by sustained daily supplementation with beta-carotene. However, after two daily treatments with 50 mu M beta-carotene intracellular ROS levels were significantly reduced and there was a trend towards reduced intracellular ROS within monolayers subject to five daily treatments with 0.5 and 5 mu M beta-carotene. Prolonged supplementation with 0.1 and 0.5 mu M beta-carotene or short supplementation periods with 5 and 50 mu M beta-carotene did not alter susceptibility to H2O2. However, cultures treated daily between 3 and 8 days post confluence with 5 or 50 mu M beta-carotene exhibited enhanced LDH release, increased non-adherence and enhanced Trypan blue staining when challenged with 10 mM H2O2. In the absence of H2O2, the beta-carotene treatments were not overtly toxic to the monolayers. These results indicate that beta-carotene does not enhance antioxidant defences within Caco-2 monolayers. The enhancement of H2O2 toxicity by persistent, high doses of beta-carotene may contribute to the failure of this carotenoid to protect high risk individuals from certain degenerative conditions. (C) 1999 Elsevier Science B.V. All rights reserved.

AB - As encountered with a plethora of other natural products, the antioxidant activity of beta-carotene has been proposed as one of the mechanisms by which diets rich in this pro-vitamin A active carotenoid apparently afford chemoprevention. Here, we report the ability of beta-carotene to alter endogenous reactive oxygen levels and antioxidant defences within non-stressed 'differentiated' monolayers of an intestinal epithelial cell line (Caco-2) and to subsequently effect resistance to general oxidative insult. The differentiated monolayers efficiently absorbed beta-carotene. Between 3 and 8 days post confluence, cultures exhibited a progressive increase in antioxidant enzyme activity and a corresponding reduction to intracellular ROS levels. The profile for antioxidant enzyme activity was unaffected by sustained daily supplementation with beta-carotene. However, after two daily treatments with 50 mu M beta-carotene intracellular ROS levels were significantly reduced and there was a trend towards reduced intracellular ROS within monolayers subject to five daily treatments with 0.5 and 5 mu M beta-carotene. Prolonged supplementation with 0.1 and 0.5 mu M beta-carotene or short supplementation periods with 5 and 50 mu M beta-carotene did not alter susceptibility to H2O2. However, cultures treated daily between 3 and 8 days post confluence with 5 or 50 mu M beta-carotene exhibited enhanced LDH release, increased non-adherence and enhanced Trypan blue staining when challenged with 10 mM H2O2. In the absence of H2O2, the beta-carotene treatments were not overtly toxic to the monolayers. These results indicate that beta-carotene does not enhance antioxidant defences within Caco-2 monolayers. The enhancement of H2O2 toxicity by persistent, high doses of beta-carotene may contribute to the failure of this carotenoid to protect high risk individuals from certain degenerative conditions. (C) 1999 Elsevier Science B.V. All rights reserved.

KW - beta-carotene

KW - antioxidant

KW - reactive oxygen species

KW - membrane damage

KW - cell culture

KW - Caco-2

KW - free-radicals

KW - lipid-peroxidation

KW - cancer prevention

KW - vitamin-E

KW - supplememtation

KW - vegetables

KW - chemoprevention

KW - catalase

KW - diseases

KW - systems

U2 - 10.1016/S0304-4165(99)00212-3 |

DO - 10.1016/S0304-4165(99)00212-3 |

M3 - Article

VL - 1474

SP - 47

EP - 55

JO - Biochimica et Biophysica Acta (BBA) - General Subjects

JF - Biochimica et Biophysica Acta (BBA) - General Subjects

SN - 0304-4165

IS - 1

ER -