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Estrogen and Progestin Bioactivity of Foods, Herbs, and Spices (44247) Figures and Tables
David T. Zava, PhD, Charles M. Dollbaum, MD, PhD and Marilyn Blen
Reprinted by permission Society for Experimental Biology and Medicine
Abstract
Introduction
Methods

Results
Progestin-Mediated Down Regulation of ER as an Index of Progestin-Agonist/Antagonist Activity of Herbal Extracts
Discussion

Permission
Acknowledge
 		Table I - Alphabetical Listing of Herbs Studied
Table 2 - Herbs and Spices Containing ER Binding Components

Table 3 - Herbs and Spices Containing PR Binding Components
Figure I - Protein/Well ER (+)

Figure 2 - Protein/Well ER (-)
Figure 3 - nmoles PNP formed/min / µg protein

Figure 4 - fmoles Estrogen Receptor/ µg DNA
Figure 5 - Log Progesterone Concentration (pg/ml)

Figure 6 - PR Binding (pg/ml Pg Equivalents)
Figure 7 - Hours Post SoyMilk Ingestion

Table 1
Alphabetical Listing of Herbs Studied
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Acacia (Acacia senegal), Agrimony (Agrimonia eupatoria), Unicorn root (Aletris farinosa), Alfalfa (Medicago sativa), Aloe (Aloe vera), Anise (Pimpinella anisum), Asparagus (Asparagus officinalis), Astragalus (Astragalus membranaceus)

Barberry (Berberis vulgaris), Bay leaf (Laurus nobilis), Bayberry (Myrica cerifera), Black Cohosh (Cimicifuga racemosa), Black Current (Ribes nigrum), Blessed Thistle (Cnicus benedictus), Bloodroot (Sanguinaria canadensis), Blue Cohosh (Caulophyllum thalictroides), Blue Flag (Iris versicolor), Borage (Borago officinalis), Bryony root (Bryonia alba), Burdock root (Arctium lappa)

Calamus root (Acorus calamus), Calendula (Calendula officinalis), Caraway seed (Carum carvi), Cascara Sagrada (Rhamnus purshiana), Catnip (Nepeta cataria), Cayenne (Capsicum annum), Celandine (Chelidonium majus), Chamomile (Chamaemelum nobile), Chaste-tree (Vitex agnus-castus), Cherry, wild (Prunus avium), Chickweed (Stellaria media), Cilantro (Coriandrum sativum), Cloves (Syzygium aromaticum), Crampbark (Viburnum opulus), Cumin (Cuminum cyminum)

Damiana (Turnera diffusa), Devil's Club (Oplopanax horridum), Dock (Rumex obtusifolius), Dong quai (Angelica sinensis)

Echinacea (Echinacea purpurea), Endive (Chicorium endivia), Ephedra-Ma Huang (Ephedra distachya), Equisetum (Equisetum arvense), Evening Primrose (Oenothera biennis), Eyebright (Euphrasia officinalis)

False Unicorn (Chamaelirium luteum), Fennugreek (Trigonella foenumgraecum), Fennel (Foeniculum vulgate), Flaxseed (Linum usitatissimum)

Garlic (Allium sativum), Gelsium (Gelsium sempervirens), Gentian root (Gentiana macrophylla), Ginger root (Zingiber officinale), Ginko (Ginko bilboa), Ginseng (Panax ginseng), Ginseng, Wild Siberian (Eleutherococcus senticosus), Goldenseal (Hydrastis canadensis), Gotu Kola (Centella asiatica), Gravel root (Eupatorium purpureum)

Henna-Red (Lawsonia inermis), Hops (Humulus lupulus), Horehound (Marrubium vulgare), Hyssop (Hyssopus officinalis)

Ironwood bark (Ostrya virginiana)

Juniper berry (Juniperus communis)

Kavakava root (Piper methysticum)

Lavender (Lavandula angustifolia), Lemon Grass (Cymbopogon citratus), Licorice (Glycyrrhiza glabra), Lobelia (Lobelia chinensis), Lovage root (Levisticum officinale)

Nettle (Urtica dioica), Nutmeg (Myristica fragrans)

Oatstraw (Avena sativa), Ocatillo (Fouquieria splendens), Onion (Allium cepa), Oregano (Origanum vulgare), Oregon Grape seed (Berberis aquifolium Pursh.)

Passion fruit (Passiflora incarnata), Pau d'Arco (Tabebuia spp.), Pennyroyal (Mentha pulegium), Peony root (Paeonia officinalis), Periwinkle (Vinca), Pipsisewa (Chimaphila umbellata), Pokeweed root (Phytolacca americana), Pomegranate (Punica granatum), Poppy seed (Papaver somniferum)

Red Clover (Trifolium pratense), Red Raspberry (Rubus idaeus), Red Root (Ceanothus americanus), Rosemary (Rosmarinus officinalis), Rue (Ruta graveolens)

Saffron (Crocus sativus), Sage (Salvia officinalis), Sarsaparilla (Smilax), Sassafras (Sassafras albidum), Savory (Satureja hortensis), Saw Palmetto (Serenoa repens), Senna (Senna), Sesame seed (Sesamum indicum) Shavegrass (Equisetum arvense) sheep Sorrel (Rumex acetosella), Shephard's Purse (Capsella bursa-pastoris), Skullcap (Scutellaria lateriflora), Slippery Elm bark (Ulmus rubra), Smallage (Apium graveolens), Solomon's Seal (Polygonatum officinale), St. John's Wort (Hypericum perforatum), Strawberry leaf (Fragaria vesca), Sumac (Rhus glabra)

Tansy (Tanacetum vulgate), Thyme (Thymus vulgaris), Turmeric (Curcuma longa)

Uva Ursi (Arctostaphylos uva-ursi)

Valerian (Valeriana officinalis), Vervain (Verbena officinalis), Vitex-chaste tree (Agnus canthus)

Wild Pansy (Viola tricolor), Wild Yam (Dioscorea villosa), Wormwood (Artemisia absinthium)

Yarrow (Achillea millefolium), Yellow dock (Rumex crispus), Yerba Mansa (Anemopsis californica), Yucca (Yucca Spp.)

Table 2
Herbs and Spices Containing ER Binding Components
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Micrograms of Estradiol Equivalents/200 cc or 2 g Dry Herb

Soy milk

 8/200cc Yucca 0.5/2g
Licorice 4/2 g Turmeric 0.5
Clover (red) 3 Hops 0.5
Mandrake 3 Verbena 0.5
Bloodroot 2 Yellow dock 0.5
Thyme 2 Sheep sorrel 0.5

Soy milk, herbs, and spices were extracted and tested for their ER-binding capacity as described in the Methods. Soy milk and 11 of the herbs and spices with the highest activity are listed and expressed in total estradiol binding equivalents per 200 cc of soy milk or 2 g of dried herb. Values were rounded to the nearest 0.5 micrograms of estradiol binding equivalents. Results are representative of at least two other assays.
 

Table 3
Herbs and Spices Containing PR Binding Components
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Micrograms of Progesterone Equivalents/2 g Dry Herb
Bloodroot 100 Thyme 4
Ocotillo 8 Calamus rt. 3
Mandrake 8 Red clover 3
Oregano 8 Goldenseal 3
Damiana 6 Licorice 3
Pennyroyal 5 Mistletoe 3
Verbena 5 Cumin 2
Nutmeg 4 Fennel 2
Turmeric 4 Camomille 2
Yucca 4 Cloves 2

Herbs and spices were extracted and tested for their PR-binding capacity as described in the Methods. The 20 herbs and spices with the highest activity are listed and expressed in progesterone-binding equivalents. Values were rounded to the nearest microgram of progesterone-binding equivalents. Results are representative of at least two other assays.

Figure1 Protein/Well ER (+)
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Figure2 Protein Well ER (-)
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Figure1 Protein Well Effects of ER-binding herb extracts on growth of ER(+) breast cancer cells

Figure 1 Effects of ER-binding herb extracts on growth of ER (+) breast cancer cells in vitro. Cells were grown in 96-well plates for 9 days in absence (control) or presence of estradiol, estradiol plus hydroxytamoxifen, coumestrol, or herbal extracts (1/500 dilution). Growth was assessed by measuring protein directly in the wells as indicated in the Methods.

Figure2 Protein Well Effects of ER-binding herb extracts on growth of ER(-) breast cancer cells

Figure 2 Effects of ER-binding herb extracts on growth of ER (-) breast cancer cells in vitro. Cells were grown in 96-well plates for 7 days in absence (control) or presence of estradiol, estradiol plus hydroxytamoxifen, coumestrol, or herbal extracts (1/500 dilution). Growth was assessed by measuring protein directly in the wells as indicated in the Methods.
 

Figure 3 - nmoles PNP formed/min / µg protein
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Figure 4 - fmoles Estrogen Receptor/ µg DNA
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Figure3 graphic - Induction of alkaline phosphatase by progesterone plus RU486 and PR binding herbal extracts

Figure 3 Induction of alkaline phosphatase by progesterone plus RU486, and PR binding herbal extracts, with and without progesterone. Near confluent cells in 96-well plates were exposed to increasing doses of progesterone, 10-9M progesterone plus 10-7MRU486 or 1/500 dilution of herbal extracts (as indicated) without or with 10-9M progesterone. After 48 hr, cells were fixed and alkaline phosphatase measured directly in the plate as indicated in Methods

Figure4 Graphic - Down regulation of ER by progesterone and herbal extracts in the absence and presence of RU486

Figure4 Down regulation of ER by progesterone and herbal extracts in the absence and presence of RU486. Near confluent cells in 24-well plates were exposed to progesterone and herbal extracts, without or with 10-9M RU486. After 48 hr. ER wa measured directly in the plate by incubating cells with 2nM [125l]-estradiol. Following incubation, the cells were removed from the plate with trypsin and the radioactive binding of [125l]-estradiol was meansured in nuclei as decribed previously. (11)

Figure 5 - Log Progesterone Concentration (pg/ml)
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Figure5 - Log Progesterone Concentration (pg/ml) picture of

Figure5 - Induction of alkaline phosphatase in T47D breast cancer cells with extracts of saliva spiked with increasing doses of progesterone. Pooled saliva was charcoal treeated then spiked with increasing concentrations of progesterone as indicated. Saliva was then extracted in DEE, reconstituted in growth media and then incubated with cells for 48 hr. Alkaline phosphastase was then determined as indicated in the Methods

Figure 6 - PR Binding (pg/ml Pg Equivalents)
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PR Binding (pg/ml Pg Equivalentspicture of

Figure6 - Induction of alkaline phosphatase in T47D breast cancer cells with extracts of saliva from premenomausal women and women taking progesterone, MPA, and wild-yam (diosgenin-containing) herbs. Saliva was processed as described in Figure 5 and incubated with cells for 48 hr. Alkaline phosphastase was measured as described above.

Figure 7 - Hours Post SoyMilk Ingestion
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Figure7 - Phytoestrogen content of saliva following soy milk consumption

Figure 7 Phytoestrogen content of saliva following soy milk consumption. Volunteers drank 200cc of soy milk and collected saliva samples at hourly intervals over 24 hr. Saliva was processed and assayed for estrogen receptor binding components and for estradiol by conventional radioimmunoassay as described in the Methods Results are expressed in picograms (pg) of estradiol binding equivalents per ml of saliva.

Permission
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Permission granted by the publisher and authors:
Copyright © 1998 The Proceedings of the Society for Experimental Biology and Medicine 1998 Mar;217(3):369-78
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