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【DNA microarray-based gene expression profiling of estrogenic chemicals】

· Research on plants
Plants are the primary source of nutrients that maintain the proper functioning of our bodies. From these plants such as agricultural crops, fruits and nuts, our bodies absorb carbohydrates, proteins and amino acids, as well as a variety of vegetable oils and vitamins. However, in our daily lives, there are also numerous crude drugs that are commonly known in the world such as herbal medicines. In Japan, there are herbal medicines that contain various active ingredients such as those found in the Chinese herbal medicines, the Ayurveda herbs of India, as well as the Aroma medicinal herbs found in western countries. In addition to the pharmaceuticals sold in drugstores, there are products including cosmetics and health foods, many of which contain raw ingredients derived from plants such as medicinal herbs.
So, what is the difference between pharmaceuticals and the traditional Chinese herbal remedies and herbal medicines?
The substances of the drugs prescribed by doctors as well as their effects have been scientifically validated, with the governmental agencies confirming to some extent the safety and effect these drugs before allowing them to be sold in the market. However, there are a number of medicinal herbs and herbal medicines found in nature that have been discovered and used by our ancestors throughout the ages whose effects cannot be proven or explained scientifically. Many of the useful substances found in plants such as these medicinal herbs are comprised of substances that resemble the signaling compounds of human hormones, having functional properties that cannot be elucidated even using present day state-of-the-art scientific and technological methods. A signaling compound is one that transmits various instructions throughout the genetic system, but differs slightly from substances such as nutrients, because even trace amounts of signaling substances on the order of 1 picomole (pM) can have a significant effect upon the human body
As a result of our collaborative research with the renowned National Institute of Advanced Industrial Science and Technology (AIST), which is engaged in the development of cutting-edge advanced technologies in the city of Tsukuba in Ibaraki prefecture in Japan, we have been using advanced genetic technologies to explore areas previously unknown to develop a genetic analysis method that is a step ahead of the rest of the world.
The key role of the signaling compounds made by plants is facilitated by the plant pigments also known as polyphenols, organic chemicals characterized by the presence of large multiples of phenol structural units. Plant-derived polyphenols are said to be often found in red wines, but it is well known that polyphenols can be commonly found in other sources such as health foods and dietary supplements, including isoflavones (found in phytoestrogenic plants) and the currently popular equol (an isoflavandiol estrogen). In addition, many polyphenols contain anthocyanins (a type of flavonoid) found in sources such as blueberries, which are said to have a therapeutic effect on the eyes, and these active substances can be found in pomegranates, which are known to alleviate menopausal disorders. Products promoting the beneficial effects of polyphenols are widely advertised on TV, but there remains an infinite number of commercial applications for these substances.
Polyphenol, because of its chemical structure and features, is remarkably similar to that of the signaling substance (estrogen) that regulates gene expression and influences physiological processes; for example, by transmitting signals during pregnancy from the pregnant mother to the baby in her womb to facilitate growth of the fetus. Throughout the life of both males and females, even when the human body has fully developed, estrogen dramatically influences diverse physiological processes and regulates gene expression in target tissues such as the brain, heart, liver, breast, uterus, bones, blood vessels, skin, ovaries, and the hypothalamus to maintain the proper functioning of the human body.
However, once the growth phase of the human body has ended and a person reaches a certain age, the secretion of hormones sharply decreases. As a result, plant-derived polyphenols can be considered as a substitute for estrogen, and when ingested have to some extent the effect of compensating for the decrease in the secretion of estrogen to relieve symptoms associated with aging and poor physical condition.
As a result, polyphenols are considered as “phytoestrogens”. In addition, these polyphenols contain telomeres, a mechanism that has the effect of inhibiting the life span of organisms other than plants, and manages the lifespan of human cells. Telomeres are structures that are found at the ends of chromosomes. They protect chromosomes from damage and shortening, which increasingly occurs as a human ages. Longer telomere structures are associated with healthy cell replication and longevity as they prolong the life of cells. Thus, polyphenols possess antiaging properties. Due to the estrogenic effect of these polyphenols, we believe that it is possible to extend our average life expectancy to 80 or 90 years old, and make it possible to remain youthful practically forever.
So, what is the effect of these polyphenols that are so similar to estrogen? Although we have only just started to study various polyphenols, we expect this new research to discover new plant functions such as anti-aging properties.

· Action of the signaling substance “estrogen”

Estrogen transmits genetic signals directly to such tissues as the brain, heart, liver, breasts, uterus, bones, blood vessels, skin, ovaries, and hypothalamus. Mainly, in women after the onset of menopause due to the associated deficiency in the secretion of estrogen, they are likely to develop such physical disorders such as osteoporosis, myocardial infarction, and hypertension. In addition, it has been reported that estrogen is effective at enabling the improved functioning of the myocardium (striated muscle cells), liver (stem cells), bones (osteoblasts), blood vessels (endothelial cells), skin (fibroblasts), and other tissues, brain functioning for diseases such as Alzheimer’s type of dementia, as well as a positive effect on the thyroid, pancreas, liver, corticoids (a group of steroid hormones) and other immune systems.

Estrogen has multifaceted effects on the hypothalamus that regulate a number of homeostatic functions including reproduction, temperature, energy balance, stress, and motivated behaviors. Estrogen targets all of the major hypothalamic neuroendocrine and autonomic cellular groups to activate multiple signaling pathways, which includes the sympathetic and parasympathetic nervous systems and endocrine functions.
The major divisions of the brain are the forebrain (prosencephalon), midbrain (mesencephalon), and hindbrain (rhombencephalon). The forebrain includes the cerebrum (telencephalon), and consists of two subdivisions called the telencephalon and diencephalon. The diencephalon region of the forebrain relays sensory information and connects components of the endocrine system with the nervous system. The diencephalon regulates a number of functions including autonomic, endocrine, and motor functions. The components of the diencephalon include the thalamus, hypothalamus, and pineal gland. The hypothalamus acts as the control center for many autonomic functions including respiration, blood pressure, and body temperature regulation. This endocrine structure secretes hormones that act on the pituitary gland to regulate biological processes including metabolism, growth, and the development of reproductive system organs. As a component of the limbic system, the hypothalamus influences various emotional responses through its influence on the pituitary gland, skeletal muscular system, and autonomic nervous system. Estrogen has the most important role in maintaining the life of cells through the hypothalamus, which acts as the control center to regulate a number of automatic behavior necessary for survival including the autonomic, endocrine, and motor functions. Specifically, it control those functions that govern blood pressure, blood flow, body temperature, body fluid, digestion, absorption, excretion, and reproduction.
For example, this would include the thirst response by stimulating the hypothalamus to make the mouth feel dry, as well as regulating blood sugar levels, and adjusting blood pressure, thereby controlling the contraction of blood vessels and the secretion of hormones such as adrenaline. It is also associated with diuretic activity, and the regulation of blood glucose levels that are controlled by signals transmitted via the liver emitted from the hypothalamus. In addition, the hypothalamus is the autonomic control center of visceral functions such as the regulation of body temperature, and the release of hormones by the pituitary and pineal glands to help regulate autonomic functions such as sleep. Unlike other areas of the brain such as the cerebral cortex, the hypothalamus is always in control of the body, and estrogen, which is present in all organs of the body, is a hormone intimately involved how the hypothalamus acts as the control center.

· New analytical method
Scinet Co., Ltd. has succeeded in identifying in those human genes that receive genetic signaling information transmitted via the hormone estrogen. The new analysis method utilizing response genes (information receptor genes) to investigate the effects of estrogen in the human body was developed in collaboration with the National Institute of Advanced Industrial Science and Technology (AIST), has been patented, and is currently being used in research just started on the signaling information of hormones found in the human body.
Research in this field remains largely unexplored. But, this analysis method is the first in the world to analyze the intensity of estrogenic activity in the important medicinal substances found in plants. Also, this in vitro method makes it possible to confirm the molecular bonds known as receptors in endocrine system cells whose function may be altered or damaged due to substances such as endocrine disrupting chemicals (EDCs) at a sensitivity greater than 1000 times over that of in vivo animal experimental methods.

(Comparison of new DNA chip and old analysis methods)

• Development history
• Patent acquisition
• Research on plants
• New analytical method
• Discovery of “silent estrogen”
• The effect on men and women during menopause
• (Necessity as an alternative method to animal experimentation and to address environmental issues)