Witold Mazur

Phyto-oestrogens and Western Diseases

Incidences of breast, colorectal and prostate cancer are high in the Western world compared to countries in Asia. We have postulated that the Western diet compared to the semivegetarian diet in some Asian countries may alter hormone production, metabolism or action at the cellular level by some biochemical mechanisms. Our interest has been focused on two groups of hormone-like diphenolic phyto-oestrogens of dietary origin, the lignans and isoflavonoids abundant in plasma of subjects living in areas with low cancer incidence. The precursors of the biologically active compounds detected in man are found in soybean products, whole-grain cereal food, seeds, and berries. The plant lignan and isoflavonoid glycosides are converted by intestinal bacteria to hormone-like compounds. The weakly oestrogenic diphenols formed influence sex-hormone production, metabolism and biological activity, intracellular enzymes, protein synthesis, growth factor action, malignant cell proliferation, differentiation, cell adhesion and angiogenesis in such a way as to make them strong candidates for a role as natural cancer-protective compounds. Their effect on some of the most important steroid biosynthetic enzymes may result in beneficial modulation of hormone concentrations and action in the cells preventing development of cancer. Owing to their oestrogenic activity they reduce hot flushes and vaginal dryness in postmenopausal women and may to some degree inhibit osteoporosis, but alone they may be insufficient for complete protection. Soy intake prevents oxidation of the low-density lipoproteins in vitro when isolated from soy-treated individuals and affect favourably plasma lipid concentrations. Animal experiments provide evidence suggesting that both lignans and isoflavonoids may prevent the development of cancer as well as atherosclerosis. However, in some of these experiments it has not been possible to separate the phyto-oestrogen effect from the effect of other components in the food. The isoflavonoids and lignans may play a significant inhibitory role in cancer development particularly in the promotional phase of the disease, but recent evidence points also to a role in the initiation stage of carcinogenesis. At present, however, no definite recommendations can be made as to the dietary amounts needed for prevention of disease. This review deals with all the above-mentioned aspects of phyto-oestrogens.

Isoflavonoids and lignans in legumes: nutritional and health aspects in humans

Abstract Dietary factors are considered important environmental risk determinants for Western diseases. Studies have revealed beneficial or protective effects of the consumption of legumes with regard to hypercholesterolaemia and coronary heart disease, obesity, diabetes mellitus, and menopause. During the last decade attention has been focused on soy and soybean products. Several constituents have been isolated: isoflavones, phytosterols, protease inhibitors, inositol hexaphosphate, and saponins. Our interest concentrates on hormone-like bisphenolic phytoestrogens of dietary origin, the lignans and isoflavonoids. Their glycosides, converted by gut bacteria to mammalian derivatives with weak estrogenic and antioxidative activity, originate in leguminous seeds. We developed an isotope dilution gas chromatography-mass spectrometry method for quantitative determination of the isoflavones, formononetin, biochanin A, daidzein, genistein and coumestrol, and the lignans secoisolariciresinol and matairesinol, in food samples. We measured the four isoflavonoids and coumestrol, and, for the first time, the two lignans in 52 leguminous seeds and found high concentrations of isoflavonoids (0–1853.35 mg/kg; 0–7.3 mmol/kg dw) but lower amount of lignans (0–15.85 mg/kg; 0.05 mmol/kg dw). The highest plasma levels of their metabolites are found in individuals living in countries or regions with low cancer and cardiovascular disease incidence and these are probably sufficient to influence intracellular enzymes, protein synthesis, growth factor action, malignant cell proliferation, differentiation, and angiogenesis. Leguminous seeds, therefore, in respect to their abundant concentrations of phytoestrogens, are strong candidates for a role as natural cancer-protective food.

11 Phytoestrogen content in foods

Plants abound in essential phytochemicals produced for their various vital functions. The same compounds seem also to be crucial for human health and disease. Recent human epidemiological and laboratory animal and cell studies on cancer and heart disease have highlighted the phytoestrogens--naturally occurring principles that share with steroidal oestrogens an ability to activate oestrogen receptors. The best known non-steroidal phytoestrogens include the isoflavones daidzein, genistein, formononetin and biochanin A, the coumestan coumestrol, and the lignans secoisolariciresinol and matairesinol. Acknowledging the potentially chemoprotective role of these non-nutrients, we have quantified all biologically important isoflavonoids and lignans in cereals, oilseeds and nuts, legumes, vegetables, fruits, berries and beverages such as tea, coffee and wine. In this chapter, we present a review of our studies on staple plant foods, indicating that plants contain, besides a wide range of chemicals with a number of biological properties, biologically active phytoestrogens--precursors of hormone-like compounds found in mammalian systems.

Naturally occurring oestrogens in food

Edible plants contain a number of natural compounds which mimic the biological effects of oestrogens by virtue of their ability to bind to and activate the nuclear oestrogen receptors. These hormone-like diphenolic phyto-oestrogens of dietary origin include isoflavonoids, coumestans and lignans. Our interest in these phyto-oestrogens derives from the results of epidemiological studies on diet and Western diseases including hormone-dependent cancers as well as coronary heart disease. Incidences of the diseases in question are lower in peoples of Asia compared to inhabitants of industrialized American and European countries. Using isotope dilution gas chromatography-mass spectrometry method we have identified and measured in foods the precursors of the biologically active compounds detected in plasma of subjects living in areas with low cancer incidence. Biochanin A, formononetin, daidzein, genistein, and coumestrol, and the lignans matairesinol and secoisolariciresinol have been found to possess oestrogenic, anti-oestrogenic, antioxidative, antiviral, antibacterial, insecticidal or fungistatic properties and they have been shown to be antiproliferative in relation to many types of tumours in cell culture. We report quantitative results for these plant oestrogens measured in soybeans and other legumes, oilseeds and nuts, grain and cereals, berries and fruits, cruciferous, allium and other vegetables, and beverages such as tea and coffee.

Lignan and isoflavonoid concentrations in tea and coffee.

Tea is a beverage consumed widely throughout the world. The existence in tea of chemopreventing compounds possessing antimutagenic, anticarcinogenic and antioxidative properties has been reported. High intakes of tea and foods containing flavonoids have recently been shown to be negatively correlated to the occurrence of CHD. However, tea may contain other compounds with similar activities. Using a new gas chromatographic-mass spectrometric method we measured lignans and isoflavonoids in samples of twenty commercial teas (black, green and red varieties) and, for comparison, six coffees. Both unbrewed and brewed tea were investigated. The analysis of the teas yielded relatively high levels of the lignans secoisolariciresinol (5.6-28.9 mg/kg; 15.9-81.9 mumol/kg) and matairesinol (0.56-4.13 mg/kg; 1.6-11.5 mumol/kg) but only low levels of isoflavonoids. Because the plant lignans, as well as their mammalian metabolites enterolactone and enterodiol, have antioxidative properties and these mammalian lignans occur in high concentrations in plasma, we hypothesize that lignan polyphenols may contribute to the protective effect of tea on CHD.

Phyto-oestrogen content of berries, and plasma concentrations and urinary excretion of enterolactone after a single strawberry-meal in human subjects

Quantitative data on phyto-oestrogen, particularly lignan, content in edible plants are insufficient. We, therefore, measured isoflavonoids and lignans in nine edible berries using an isotope dilution gas chromatography-mass spectrometry method for foods and found substantial concentrations of the lignan secoisolariciresinol (1.39-37.18 mg/kg DM), low amounts of matairesinol (0-0.78 mg/kg DM) and no isoflavones. To determine pharmacokinetics and urinary excretion pattern of the mammalian lignan enterolactone derived from plant lignans, a study with human subjects was conducted. Five healthy women and two men consumed, after a 72 h period of a phyto-oestrogen-free regimen, a single strawberry-meal containing known amounts of plant lignans. Basal and post-meal blood and urine samples were collected at short intervals. The samples were analysed using time-resolved fluoroimmunoassay of enterolactone. The meal increased plasma concentration of enterolactone after 8-24 h and in urine in the 13-24 h and 25-36 h urine collections. High individual variability of the metabolic response was observed. Enterolactone excreted in the urine collected throughout the 48 h post-meal yielded on average 114% of the plant lignans consumed. It is concluded that berries containing relatively high concentrations of plant lignans contribute to plasma and urinary levels of mammalian enterolactone in human subjects.

Inhibitory effects of soy and rye diets on the development of Dunning R3327 prostate adenocarcinoma in rats

Dunning R3327 PAP prostate tumors were transplanted in 125 rats, the rats were divided into five groups, and tumor development was examined for 24 weeks during treatment with diets containing 33% of soy flour (SD), rye bran (RB), heat‐treated rye bran (HRB), or rye endosperm (RE).

Phyto-oestrogen database of foods and average intake in Finland

Information on phyto-oestrogen intake in various populations has been scanty until now, primarily because data on the content of these compounds in foods were lacking. We report here on expansion of the Finnish National Food Composition Database (Fineli) with values for the plant lignans matairesinol and secoisolariciresinol and the isoflavones daidzein and genistein. The values, expressed as aglycones, were based on food analyses (mainly GC-MS) or imputed from analytical data for 180 foods for lignans and 160 foods for isoflavones; additionally, over 1000 values were derived from the recipe database of Fineli. Average intake of these phyto-oestrogens was calculated using food consumption data of the National Dietary Survey FINDIET 1997, which was carried out in a random sample of the adult population in five areas in Finland. The dietary data were collected by 24 h recall =2862). The mean lignan intake was 434 (standard deviation (SD) 1575) microg/d and the mean isoflavone intake was 788 (SD 673) microg/d. Women had a higher lignan density (microg lignans/MJ) in their diet than men (P<0.05). Men had a higher mean daily isoflavone intake, 902 (SD 368) microg, than women, 668 (SD 963) microg (P<0.05). The sources of lignans were many: seeds, cereals, fruit, berries and vegetables. The main sources of isoflavones appeared to be processed meat products/sausages containing soya as an ingredient, and legumes as such. The average intake of lignans and isoflavones in Finland seems to be low, but intake varies throughout the population.

Consumption of wholemeal rye bread increases serum concentrations and urinary excretion of enterolactone compared with consumption of white wheat bread in healthy Finnish men and women.

Rye is an important source of plant lignans in Finland. In the present crossover trial we wanted to study the effect of rye bread as part of the usual diet on serum and urine enterolactone (ENL) concentrations in healthy volunteers. Eighteen men aged 43 (sem 2.0) years and twenty-one women aged 43 (sem 1.6) years consumed wholemeal rye bread and white wheat bread in random order for 4 weeks. The bread periods were separated by a 4 week wash-out period. The breads provided at least 20% of the daily energy intake. The mean intakes of rye bread were 219 (sem 14.6) and 162 (sem 5.3) g/d and those of wheat bread were 200 (sem 9.6) and 153 (sem 5.8) g/d for men and women respectively. Blood samples were collected from all subjects and three 24 h urine samples were collected from ten men and twelve women at the end of both bread periods for the determination of serum concentration and urinary excretion of ENL. The mean serum ENL concentrations in both men and women at the beginning of baseline period and at the end of the rye-bread period remained constant and were significantly higher than those at the end of the wheat-bread period. Correspondingly, daily urinary ENL excretion increased significantly during the rye-bread period compared with the wheat-bread period and was 5- and 10-fold higher in men and women respectively in comparison with the amount of plant lignan precursors measured in the rye bread. These data indicate the presence of other precursors for ENL in rye which are not detected by the current method of measuring plant lignans in food. The possible role of fibre in enhancement of the formation of mammalian lignans from their plant precursors in the gut also remains to be determined.

Content of nutrients and lignans in roller milled fractions of rye

Rye grain was roller milled into six flour fractions (607 g kg-1), a short (341 g kg-1) and a bran (52 g kg-1) in a B???hler laboratory mill. In the different flours (F1-F6) a progressive increase in ash (4-13 g kg-1), crude protein (39-87 g kg-1) and crude fat (5-17 g kg-1) concentration was found and a corresponding decrease in starch (853-699 g kg-1) concentration. Total dietary fibre (55-109 g kg-1) constituents and the lignans, matairesinol (0???05-0???22 mg kg-1) and secoisolariciresinol (0???21-0???38 mg kg-1), showed a different distribution with a maximal concentration in F5. The short and bran had higher concentrations of ash, crude protein, crude fat, dietary fibre components and lignans but a lower concentration of starch than the flours. Compared to the short, the bran contained more of all constituents analysed except mixed-linked beta-glucan and starch. A very high proportion of total dietary fibre components of the rye, such as arabinoxylan (72%), cellulose (76%) and Klason lignin (79%) as well as of matairesinol (87%) and secoisolariciresinol (73%), was found in the short and bran together. ??? 1997 SCI.