Ovulation Block by &lt;I&gt;Pueraria mirifica&lt;/I&gt;: A Study of Its Endocrinological Effect in Female Monkeys

Abstract. The aim of this study was to evaluate the estrogenic activity of tuberous samples of wild, phytoestrogen-rich Pueraria mirifica collected from 28 out of 76 provinces of Thailand by MCF-7 proliferation assay. The plant extracts were administered to MCF-7, ERα positive human mammary adenocarcinoma cell cultures, for 3 days at dosages of 0.1, 1, 10, 100 and 1,000 µg/ml and were compared with 17β-estradiol at concentrations of 10 -12 -10 -6 M. The mean P. mirifica population at 1 µg/ml exhibited significant proliferation. Two plant samples exhibited levels of proliferation in MCF-7 that were similar to 17β-estradiol. The mean P. mirifica populations at 100 and 1,000 µg/ml exhibited significant cytotoxicity in MCF-7. Analysis of the estrogenic activity of puerarin, representative of major isoflavonoids in P. mirifica tubers, revealed proliferation in MCF-7 only at the highest dose (10 -6 M) that was 10 2 -10 5 times less active than 17β-estradiol. Puerarin and 17β-estradiol at concentration of 10 -12 -10 -6 M exhibited no cytotoxicity in MCF-7. Key words: Isoflavonoid, MCF-7 proliferation assay, Phytoestrogen, Pueraria mirifica, Puerarin (J. Reprod. Dev. 54: [63][64][65][66][67] 2008) he plants in genus Pueraria are largely distributed in Asia and have a long history of ethnopharmacological applications [1]. Pueraria mirifica Airy Shaw et Suvatabhandu, family Leguminosae, is an indigenous Thai herb. The potent estrogenic activity of the plant has been proven in a 1-year human clinical trial of 4 menopausal volunteers at Ramathibodi Hospital, Faculty of Medicine, Mahidol University, Bangkok, Thailand. An average dose of 100 mg of tuberous powder per day was effective for long-term treatment of menopausal symptoms [2], and toxicological tests showed that it did not have significant adverse effects in animals and human volunteers. An acute toxicity test of P. mirifica treatment at a dose of 16 g/kg BW using rats did not reveal any adverse effects [3]. However, P. mirifica did initiate adverse effects such as anemia, during subchronic treatment for 3 months at doses of 100 and 1,000 mg/kg BW [4]. Haemotological adverse effects were confirmed in a 6-month chronic toxicity test at a dose of 250 mg/kg BW [5]. Treatment with the plant extract at high doses 600 and 800 mg/kg BW, for 30 days induces micronucleus formation in male rats [6]. However, pretreatment of weanling rats with P. mirifica powder at a dose of 10, 100 and 1,000 mg/kg BW for 28 days resulted in a dose-dependent prevention of mammary tumors induced by 7, 12-dimethylbenz (a) anthracene (7, 12-DMBA) [7].The phytoestrogens of P. mirifica exhibit the following responses: dose-dependent estrogenic effects on the reproductive system, initiated vaginal cornification, increased uterine weight with decreased follicle stimulating hormone (FSH) and luteinizing hormone (LH) serum levels in ovariectomized rats [8], increased length of the follicular phase and total menstruation cycle [9], decreased estrogen, progesterone, FSH, and LH concentrations at the do...

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confidence: 99%

Determination of the Estrogenic Activity of Wild Phytoestrogen-rich Pueraria mirifica by MCF-7 Proliferation Assay

Cherdshewasart

Traisup

Picha³

2008

“…Feeding PM powder suspended in water to ovariectomized rats induces proliferation of the vaginal epithelium and uterus endometrium and reduces the levels of serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) [7,8]. Reduction of the levels of FSH and LH has also been observed in adult cyclic and aged menopausal female monkeys [9][10][11][12]. PM suppresses folliculogenesis and ovulation in adult cyclic female monkeys after a single or long-term feeding [9,10].…”

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confidence: 99%

“…Reduction of the levels of FSH and LH has also been observed in adult cyclic and aged menopausal female monkeys [9][10][11][12]. PM suppresses folliculogenesis and ovulation in adult cyclic female monkeys after a single or long-term feeding [9,10]. Intake of crude P. mirifica powder can relieve climacteric symptoms, such as hot flashes, frustration, sleep disorders and skin dryness in post-menopausal women [13].…”

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confidence: 99%

Assessment of Fertility and Reproductive Toxicity in Adult Female Mice after Long-Term Exposure to Pueraria mirifica Herb

Jaroenporn

Malaivijitnond

Wattanasirmkit

et al. 2007

Self Cite

Abstract. The present study investigated the effects of long-term administration of Pueraria mirifica (PM) at non-toxic doses on the ovarian function and fertility of adult female mice based on evaluation of hematological and biochemical parameters [1]. Female mice were divided into 4 groups (36 mice/ group). Groups 1-3 were orally treated with a dose of 0 (PM-0), 10 (PM-10) or 100 mg/kg BW/day PM (PM-100), and group 4 was subcutaneously injected with 200 µg/kg BW/day of synthetic estrogen diethylstilbestrol (DES). The treatment schedule was separated into treatment and post-treatment periods. The duration of each period was 8 weeks. The PM-10 mice exhibited regular estrous cycles, while the PM-100 and DES treatments induced prolonged estrous cycles. Although no changes were observed in the uterus and ovary weights of the mice after the PM-100 and DES treatments, hyperplasia of the uterine endothelium and a decrease in the number of growing ovarian follicles were detected. The changes in the ovarian histologies of the PM-100 and DES mice were related to reductions in the levels of LH and FSH, which subsequently caused a decrease in mating efficiency. Once the PM mice were able to copulate, they were capable of successfully becoming pregnant and mothering offspring. No abnormalities were observed in the external morphologies and reproductive organ weights of the 50-day-old offspring. In conclusion, our results suggest that long-term exposure to 100 mg/kg BW of PM has adverse effects on the mating efficiency and reproduction of adult female mice and that administration of 10 mg/kg BW of PM does not induce any changes in the hypothalamic-pituitary-ovarian-uterine axis.

“…(PM), a Thai herbal plant, belongs to family Leguminosae [5][6][7]. The PM root has been analyzed and found to contain many phytoestrogens, including isoflavone genistein, daidzein, and coumestrol [5].…”

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confidence: 99%

“…PM influences the reproductive system by reducing serum gonadotropin levels and increasing uterine weight and vaginal cornification in rats [8]. Our previous studies have found that daily treatment with PM induces suppression of serum follicle stimulating hormone (FSH), l u t e i n i z i n g h o r m o n e ( L H ) , e s t r a d i o l , a n d progesterone in adult female cynomolgus monkeys (Macaca fascicularis) in a dose-dependent manner [6], but also that treatment with a single dose of PM has no effect on these hormones [7]. Reduction of serum gonadotropin and sex steroid hormone levels from long-term PM treatment implies disturbance of folliculogenesis and ovulation in monkeys.…”

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confidence: 99%

Assessment of Urinary Gonadotropin and Steroid Hormone Profiles of Female Cynomolgus Monkeys after Treatment with Pueraria mirifica

Trisomboon

Malaivijitnond

Cherdshewasart

et al. 2007

Self Cite

Abstract. This study investigated the changes in the urinary hormone levels of female monkeys (Macaca fascicularis) after single-dose and long-term treatments with Pueraria mirifica (PM). The monkeys were separated into 3 groups (n=3) and orally treated with 10, 100, or 1,000 mg of PM in each group. Two series of experiments were performed. In the first series of experiments, the monkeys were orally treated with a single dose of PM. The experimental schedule was divided into a one menstrual cycle pretreatment period and a two menstrual cycle post-treatment period. In the second series of experiments, the monkeys were orally treated daily with PM for 90 days. The experiment schedule was divided into a one menstrual cycle pretreatment period, a three menstrual cycle treatment period, and a two menstrual cycle post-treatment period. Urinary samples were collected daily and assayed for the FSH, LH, estradiol, and progesterone levels. The results showed that there were no changes in the FSH, LH, estradiol, and progesterone levels after treatment with a single dose of 10, 100, or 1,000 mg of PM or after daily treatment with 10 mg of PM for 90 days compared with the levels observed during the pretreatment period. Daily treatment with 100 mg and 1,000 mg of PM for 90 days only produced a clear reduction in the urinary FSH levels. This suggests that changes of urinary FSH levels can be considered an indicator for study of estrogenic effects on hormonal levels in female monkeys.