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Short-term tests of estrogenic potential of plant stanols and plant stanol esters

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Author: Turnbull, D. · Frankos, V.H. · Leeman, W.R. · Jonker, D.
Type:article
Date:1999
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Regulatory Toxicology and Pharmacology, 2 I, 29, 211-215
Identifier: 235061
doi: doi:10.1006/rtph.1999.1290
Keywords: Nutrition · Fatty acid ester · Phytosterol · Sitostanol · Sitosterol · Body weight · Cell proliferation · Chemical structure · Cytotoxicity · Drug activity · Drug screening · Estrogen activity · Food intake · Priority journal · Structure analysis · Analysis of Variance · Animals · Breast · Cell Division · Dihydrotestosterone · Esters · Estrogens, Non-Steroidal · Female · Humans · Isoflavones · Phytoestrogens · Phytosterols · Plant Preparations · Rats · Rats, Wistar · Tumor Cells, Cultured · Uterus

Abstract

To test for potential estrogenic activity of plant stanols and plant stanol esters, two short-term tests were performed. These were the E-screen test, which measures a substance's ability to induce proliferation of estrogen-responsive human breast adenocarcinoma (MCF-7) cells in culture, and an in vivo test, which measures uterotrophic activity in immature female rats fed the test substance. Four samples of vegetable oil-derived stanols (containing 88-99% stanols) were tested in the E-screen test, and one sample of wood-derived and one of vegetable oil-derived stanol fatty acid esters were tested in the in vivo test. In the E-screen test, the positive control substance, 17β-estradiol, at 100 pM, produced a statistically significant, 11.6-fold increase in cell proliferation, as measured by sulforhodamine B staining. None of the stanol preparations produced any increase in cell proliferation when tested at 1,10, and 100 μM. The highest dose of each stanol sample was associated with microscopic evidence of cytotoxicity and crystalline precipitation in the culture dishes. In the in vivo test, the positive control compound, diethylstilbestrol, produced a significant, dose- related increase in absolute and relative uterus weight in young female rats (17 days old at the start of treatment) fed the compound at 5, 10, and 20 ppb in the diet for 4 days. Neither of the two stanol ester preparations caused any significant change in absolute or relative uterus weight when fed at a concentration of 8.3% in the diet for 4 days. Thus, under the conditions of testing used, neither the free stanols nor the stanol fatty acid ester preparations showed evidence of estrogenic or uterotrophic activity.