Requirement of Metabolic Activation for Estrogenic Activity of Pueraria Mirifica

A wide range of chemicals derived from plant and human-made xenobiotics are reported to have hormonal activities. The present study was performed to examine the estrogenic effect of Kwao Keur, Pueraria mirifica (PM), that has been used as a rejuvenating folk medicine in Thailand, using recombinant yeast, MCF-7 cell proliferation and HepG2 cell transient transfection assay. In recombinant yeast assay, 0.025, 0.25, 2.5, 25,2.5 102, 2.5 103, 2.5 104 ng/ml concentrations of PM did not show any estrogenic activities, while 10-9 of 17-estradiol (positive control) showed high estrogenic activity. Estrogenic activities were induced at 2.5ng/ml to 25/ml concentrations of PM in a dose-dependent manner on MCF-7 cells and the estrogenic effect of PM was blocked by tamoxifen treatment, a well-known anti-estrogen. PM also showed estrogenic effect on human hepatoma cell line, HepG2 cells, containing estrogen receptor and luciferase reporter gene. Taken together, PM in itself may have no estrogenicity in yeast system, but it has estrogenicity in MCF-7 & HepG2 cells that have human metabolic enzymes. The results indicated that PM may require metabolic activation for estrogenic activity.

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Effect of Miroestrol and Isomiroestrol on Neuroprotective Activities

Several evidences indicate that the accumulation of beta-amyloid, the formation of neurofibrillary tangles, oxidative stress, the reduction of acetylcholine (ACh) and estrogen play important roles in the pathoetiology of Alzheimer’s disease (AD). Based on multifactorial etiology of AD, controlling pathological processes of AD via multiple targets is more efficient than inhibition of single target. Therefore, a compound with multi-target action involved in pathological processes might be a potential candidate for AD. Pueraria candollei Wall. Ex Benth var. mirifica (Airy Shaw & Suvatabandhu) Niyomdhum, known as Kwaao kheur kao in Thai, is a rich source of phytoestrogen eg. genistein, daidzein, puerarin, mirificin, miroestrol and isomiroestrol2. Moreover, miroestrol was reported for highest estrogenic potency among all known phytoestrogen. Besides, Pueraria candollei extract also was claimed to have antioxidant action, since it contains abundant of flavonoids. The aim of our study was to evaluate the effects of phytoestrogen extracted from Pueraria candollei, miroestrol and isomiroestrol, on factors related AD. The test compounds were investigated for free radical scavenging and acetylcholinesterase (AChE) inhibitory activity in vitro. The interaction between the test compounds and alpha estrogenic receptor (ERα) was also studied using molecular modeling techniques.

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Research of Puerarin and Phytoestrogens from Pueraria mirifica on Rat Osteoblast

Phytoestrogens have attracted attention for their potential in the prevention of postmenopausal osteoporosis. Recently, phytoestrogen—rich herb Pueraria mirifica has been demonstrated to possess an osteogenic effect on bone in ovariectomized rats, but its underlying cellular mechanism was not known. Here, we investigated the effects of P. mirifica extract and its major isoflavone compound, puerarin, on cell viability, cell proliferation and the expression of differentiation markers in rat osteoblast—like UMR106bcells. After exposure to 17β—estradiol (E2), genistein, Pueraria mirifica extract and puerarin, proliferation but not viability 0fUMR106 cells was markedly decreased. Quantitative real—time PCR revealed that Pueraria mirifica extract and puerarin significantly increased the mRNA expression of alkaline phosphatase (ALP) and osteoprotegerin, but not Runx2, osterix or osteocalcin. Puerarin also decreased the mRNA expression of receptor activator of nuclear factor—KB ligand, an osteoclastogenic factor, suggesting that it could induce bone gain by enhancing osteoblast differentiation and suppressing osteoclast function. Furthermore, after an exposure to high affinity estrogen receptor(ER) antagonist (ICI182780), the E2—, genistein—, Pueraria mirifica extract— and puerarin—induced upregulation of ALP expressions were completely abolished. It could be concluded that Pueraria mirifica extract and puerarin induced osteoblast differentiation rather than osteoblast proliferation in an ER—dependent manner. The present findings, therefore, corroborated the potential benefit of Pueraria mirifica extract and puerarin in the prevention and treatment of postmenopausal osteoporosis.

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Research on Pueraria mirifica Extract in Primary Baboon Osteoblasts

Phytoestrogen—rich Pueraria mirifica (PM) tuberous extract is a promising candidate for the development of anti—osteoporosis drugs for postmenopausal women, but its action has never been validated in humans or in non—human primates, which are more closely related to humans than rodents. In vitro study of non—human primate osteoblasts is thus fundamental to prepare for in vivo studies of phytoestrogen effects on primate bone. This study aimed to establish a culture system of baboon primary osteoblasts and to investigate the effects of PM extract and its phytoestrogens on these cells. Primary osteoblasts from adult baboon fibulae exhibited osteoblast characteristics in regard to proliferation, differentiation, mineralization, and Puerarin estrogen receptor expression. They responded to 17β—estradiol by increased proliferation rate and mRNA levels of alkaline phosphatase (ALP), type I collagen, and osteocalcin. After being exposed for 48 h to 100 μg/ml PM extract, 1000 nM genistein, or 1000 nM puerarin, primary baboon osteoblasts markedly increased the rate of proliferation and mRNA levels of ALP and type I collagen without changes in Runx2, osterix, or osteocalcin expression. PM extract, genistein, and puerarin also decreased the RANKL/OPG ratio, suggesting that they could decrease osteoclast—mediated bone resorption. However, neither Pueraria mirifica extract nor its phytoestrogens altered calcium deposition in osteoblast culture. In conclusion, we have established baboon primary osteoblast culture, which is a new tool for bone research and drug discovery. Furthermore, the present results provide substantial support for the potential of PM extract and its phytoestrogens to be developed as therapeutic agents against bone fragility.

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Research on formulation of Pueraria mirifica in anti-aging product

For more than 500 years, people in South East Asia have been using the tuberous root of “Pueraria mirifica (PM) or White Kwao Krua” for its useful anti-aging properties. It belongs to the family Leguminosae, subfamily Papilionoideae or the soy, bean subfamily. The mean of name mirifica in Latin it means miracle. Pueraria mirifica is most popular in Thailand, China and Japan as shown in Figure 1, owing to the referent for Thai natural product over chemical ones. The isolation and identification of estrogen from the root of Pueraria mirifica benefits are attributed to the plants generous content of phytoestrogen, including miroestrol and deoxymiroestrol which are believed to exert particularly positive effects being similar in chemical structure to human estriol, a female estrogen. Various active ingredients such as miroestrol, deoxymiroestrol, daidzein, genistin, genistein, B-Sitosterol, stigmasterol, cournestrol, pueraria, campesterol, mirificournestan, kwakhurin, and mirificine are obtained from Pueraria mirifica (Figure. 2). It has been widely used as health product for improving skin tones and muscle firmness, reducing stress, maintaining a youthful look as an anti-aging herbal food dietary supplement, improving circulation, as an effective natural herbal remedy for menopausal, prostate problems, and revitalizing human cells by replenishing depleting hormones. It is used for like antioxidative activity, aging skin, breast enlargement, anti-wrinkle agent, sagging breasts, wrinkled skin and gray hair care product in cosmetic products. Pueraria mirifica in the form of crude extract has been introduced just recently. With modern extraction technology, the extract from dried tuber roots is standardized and prepared in the form of powder and solution. The extract solution is used in cosmetic and cosmaceutical industry such as preparation of breast cream, eye gel and skin moisturizer. However, the development of Pueraria mirifica on the moisturizing gel for anti-aging is not reported.

Therefore, the purpose of this study was to formulate a moisturizing gel that consisting of Pueraria mirifica extract for anti-wrinkle agent for anti-aging product in woman.

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Efficacy Comparison of Pueraria mirifica against Conjugated Equine Estrogen

Perimenopausal women attending the Menopausal clinic of Hat Yai Regional Hospital were voluntarily recruited. The vasomotor symptoms such as hot flushes and night sweats, as well as other unpleasant symptoms, urogenital and psychological symptoms, were also assessed. Patients were voluntarily enrolled and randomly received daily 50 mg raw material of Pueraria mirifica, Group A, or daily 0.625 mg of conjugated equine estrogen (CEE) with/without 2.5 mg of medroxyprogesterone acetate (MPA), Group B, depend on nonhysterectomized/ hysterectomized condition.

Seventy-one patients were enrolled. Eleven of those were excluded for failing to complete the initial work-up and follow-up. Sixty cases were evaluated, 30 cases in Group A and 30 cases in Group B. After medication, the mean of modified Greene climacteric scale (MGCS) in Group A/Group B had decreased from 29.0/32.26 to 17.86/18.1, 12.56/9.57 and 9.9/8.16 at 1-, 3-, and 6- month respectively. The clinical satisfaction using MGCS was not statistically significant between Pueraria mirifica (Group A) and CEE with/without MPA (Group B) in the alleviation of climacteric symptoms (p-value > 0.05,). There were no statistically significant changes of three serum markers: estradiol, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) between both groups.

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Pueraria Mirifica Research Review

Pueraria mirifica is a plant which belongs to the family Papilionaceae (Leguminosae). The root is the part of the plant that has been used in Thailand for more than 700 years. It has also been called Kwao Krua and Thai Kudzu. This plant grows in forest regions in northern Thailand. Legend has touted the anti-aging properties of using this plant medicine for centuries. A palm leaf translated from Lana in 1931, discusses the use of the root, Pueraria Tuberous Root, to “make the skin smooth like a six year old child and allow you to live 1,000 years and prevent suffering from parasites, while also enhancing memory.”

Folklore goes on to say that the root is a “fountain of youth” for aged men and women. It helps to smooth wrinkled skin, supports healthy hair growth, improves eye health, sharpens the memory, increases energy and vigor, improves appetite and supports restful sleep. It’s no wonder that the Latin name of the plant, “MIRIFICA” means miracle!

In Thailand, the Ministry of Public Health is Similar to the National Institute of Health (NIH), in the United States. The Thailand Ministry of Public Health officially endorses Pueraria, due to the long history of safe use, along with modern scientific data that supports its safety and efficacy. Safety studies show that the normal therapeutic dose, provides a wide margin of safety. The Brazilian Journal of Medical and Biological Research published a study in September 2009, entitled “ The mutagenic and antimutagenic effects of the traditional phytoestrogen-rich herbs, Pueraria mirifica and Pueraria lobata.” Their conclusion was, “The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.”

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The Efficacy of Pueraria Mirifica as a Dietary Supplement

For more than 500 years, a root found in Thailand, known as “white Kwao Krua” (Pueraria candollei var. mirifica Airy Shaw et Suvat), has been described as having profound anti-aging properties.  Characterization studies carrier out since the mid-twentieth century has found that the root has unique phenol estrogenic compounds significantly, such as miroestrol, that are more potent than any other phytoestrogen found in nature. In recent years, the toxicology and efficacy of this root has been studied, including, acute, subacute, and chronic toxicology studies in animals, and human safety and efficacy studies. There is sufficient safety data for this product to be introduced into the US dietary supplement market as a new botanical product.

Breast Enhancement

A Japanese study conducted in 1999 at the School of Medicine, Saint Mariane University, Tokyo, Japan, examined data on the effect of P. mirifica root on breast enhancement. In this study, 50 healthy menstruating volunteer females, ages 20 to 49, were given between 100 to 600 mg of Pueraria mirifica powder daily orally as capsules for 7 days, two weeks after menstruation. No significant changes were seen in female hormone levels (serum estrogen, urine estrogen, and urine pregnanediol), kidney function (total urine volume, specific gravity, creatinine clearance), blood chemistries (serum total protein, total cholesterol, triglycerides, GOT, GTP, sodium, potassium chloride, calcium, and total phosphate), while blood cells (neutrophil [segmented and non-segmented], eosiniphil, basophil, lymphocyte, and monocyte),      hematocrit, hemoglobin, blood platelets, white blood cell count (WBC), or red blood cell count (RBC), between baseline and 14 days after oral intake.

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Pueraria mirifica Effects on Nerve Pain in Rats

Pueraria mirifica is a plant in the Leguminosae family, locally known in Thai as Khao-kruea-khao. This plant’s tuberous root contains phytoestrogen compounds which are similar to mammalian estrogen. Phytoestrogen comprises chemicals which inhibit inflammatory mediator. Moreover, it may help in nerve regeneration and prevent cell death as mammalian estrogen does. Therefore, this study was aimed to investigate the effects of Pueraria mirifica on neuropathic pain induced rats. The rats were divided into four groups, two control groups which are the general control (CT) and the surgical control (Sham) and two subjective groups which are the neuropathic pain induced group treated with 0.9% normal saline (SNI—N) and the neuropathic pain induced group treated with Pueraria mirifica suspension (SNI—PM). The results indicated that the neuropathic pain induced group which received Pueraria mirifica suspension (SNI—PM) showed the minimum of pain withdrawal threshold at day 5 after nerve surgery and increased to the maximum threshold closely to the baseline at 26.00 g (normal state) in the sixth week. Pueraria mirifica may decrease neuropathic pain after nerve injury. Less pain was found during the regeneration process of damaged nerves, and more movement could be enhanced, therefore, it may also promote the recovery of nerve function in rats.

For more information, please read more at: https://www.abelherb.com/effects-of-pueraria-mirifica-on-neuropathic-pain-in-rats/

Effects of Pueraria mirifica on Bone Loss in Rats

This study aimed to investigate the effects of three-month administration of crude of Pueraria mirifica, a Thai phytoestrogen containing plant, on bone loss and reproductive organs in the late middle age (7 months old) ovariectomized (OVX) and orchidectomized (ORX) rats.

After three months, rats were sacrificed and the bone mineral density (BMD) and bone mineral content (BMC) at the tibia, femur and the 4lh lumbar vertebral body were measured with a peripheral quantitative computerized tomography (pQCT) densitometry and the bone microarchitecture was determined by histological technique.

Bone loss in trabecular and cortical compartments at the various sites in axial bone (fourth lumbar vertebral body) and long bones (tibia and femur) induced by OVX in female rats or ORX in male rats was dose—dependently prevented by P. mirifica. The effects of 100 mg/kg BW/day of P. mirifica treatment on prevention of bone loss were equivalent to those of 0.1 mg/kg BW/day of EE.

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Pueraria mirifica Effects on Rats and Rabbits

Pueraria mirifica (White Kwao Keur) is an indigenous herb of Thailand which possess estrogenic—like activity. Experiments were designed to study the effects of P. mirifica on lipid profile anti endothelium-dependent vascular response in hypercholesterolemic rats and ovariectomized rabbits. Part I: 30 male Wistar rats were randomly assigned to three groups. The rats were fed normal diet serve as control group. The other 2 groups, rats were fed diet containing 1% cholesterol (cholesterol group) and l% cholesterol supplemented with P. mirifica 100 mg/kg/day (cholesterol+P. mirifica group). After 90 days, blood samples were obtained for the evaluation of total cholesterol, triglyceride, HDL-C and LDL—C. The aortas were removed, cut into ring and measured for isometric tension. Total cholesterol, LDL-C, HDL—C and triglyceride were significantly decreased in the cholesterol+P. mirifica group compared with cholesterol group (p<0.05). In addition, supplementation with 100 mg/kg/day P. mirifica significantly improved HDL/LDL ratio.

Part 2 : 20 ovariectomized (OVX) rabbits were randomly assigned to four groups. The normal rabbits were orally administered with double distilled water. The other 3 groups, OVX rabbits were orally administered with double distilled water (ovariectomized group), l7B-estradiol 4 mg/kg/day (OVX+Estrogen group) and P. mirifica 100 mg/kg/day (OVX+P. mirifica group). Blood samples were obtained every 4 weeks for analysis of blood biochemistry and serum lipid parameters. After 90 days, the aortas were measured for vascular function and pathologic examination of endothelial cell.

For more information, please read more at: https://www.abelherb.com/effects-of-pueraria-mirifica-on-isolated-aorta-in-high-cholesterol-fed-rats-and-ovariectomized-rabbits/

Research on Vaginal Atrophy Improvements after Pueraria mirifica Application

The estrogenic efficacy of topical vaginal application of Pueraria mirifica extract (PM) on the restoration of vaginal atrophy. Topical vaginal treatment with PM stimulated the maturation of the vaginal epithelium without causing systemic side effects in postmenopausal monkeys. The implication is that PM could be a safer alternative to treat vaginal atrophy in postmenopausal women.

Together with vaginal atrophy, a decrease in vaginal secretion and increase in vaginal pH also occurs, which leads to an increased incidence of vaginitis, vaginal dryness, itching, burning and irritation . Most vaginal atrophic patients complain of dyspareunia during sexual intercourse. Conjugated equine estrogens (CEE) cream, which contained a mixture of estrone, equilin, 17β-dihydroequilin,17α-estradiol and 17α-dihydroequilin at 0.625 mg/g cream, is currently the most common choice of vaginal product for the treatment of vaginal atrophy. However, the reported side effects of CEE cream include an increased occurrence of endometrial hyperplasia, endometrial stimulation, breast tenderness and uterine bleeding . Therefore, the use of synthetic phytoestrogens or an extract of phytoestrogen containing plants for the treatment of vaginal atrophy has become attractive as a potentially safer alternative . Pueraria mirifica (PM) is an endemic herb of Thailand, and its tuberous root contains a high amount of phytoestrogens.

If daily topical application of a vaginal cream containing PM extract showed a similar efficacy to that of CEE cream using vaginal tissue proliferation and the vaginal pH as markers, (ii) if vaginally administered PM and CEE creams could be absorbed through the vaginal mucosa into the blood circulation and have a systemic side effect using the decrease in plasma luteinizing hormone (LH) as an indicator and (iii) if changes in sex skin reddening in postmenopausal cynomolgus macaques can reflect the systemic effects of vaginal application of CEE and PM.

For more information, please read more at: https://www.abelherb.com/improvements-of-vaginal-atrophy-without-systemic-side-effects-after-topical-application-of-pueraria-mirifica/