Traditional Medicine (Mahuang-Tang) Improves Ovarian Dysfunction and the Regulation of Steroidogenic Genes in Letrozole-Induced PCOS Rats
Graphical abstract
Introduction
The ovaries play a central role in female reproduction and secrete sex-steroid hormones that are essential for reproductive health in women. The ovarian hormones estrogen, progesterone, inhibin, and testosterone play important roles in the menstrual cycle and fertility (Bauer and Cooper-Mahkorn, 2008). The ovaries are filled with numerous follicles and have a fluid-filled structure in which oocytes grow to maturity. The proper stepwise maturation of the ovarian follicle plays a very important role in maintaining hormonal balance in the body (Bauer and Cooper-Mahkorn, 2008; Kim et al., 2016). Estrogen is a major ovarian hormone and plays a role in follicular development due to its ability to inhibit gonadotropins such as the follicular-stimulating hormone (FSH). In addition, the rising levels of estrogen in the mid-follicular phase of the cycle result in the proliferation of granulosa cells and the maturation of follicles (Richards, 1975), and estrogen augments the aromatase reaction that creates estrogen from androgens (Taymor, 1996). Androgens are produced in the theca under the stimulation by the luteinizing hormone (LH), which can inhibit the growth effect of estrogens on granulosa cells and lead to the occurrence of atresia (Taymor, 1996).
Polycystic ovary syndrome (PCOS) involves a series of symptoms, such as irregular or missing menstrual cycles, weight gain, facial hair, etc., caused by increased androgen production in women (Hart et al., 2004). When the ovaries are simulated to produce excessive androgen hormones, especially testosterone, PCOS is triggered by one or a combination of the following: the release of excessive luteinizing hormone (LH) and high levels of blood insulin (Paradisi et al., 2001). In women, PCOS patients have an increased frequency of hypothalamic gonadotrophin releasing hormone (GnRH) pulses and consequently, an increased LH/FSH ratio (Lewandowski et al., 2011). The majority of women with PCOS have insulin resistance and/or are obese, and elevated insulin levels are an abnormal cause of the hypothalamic–pituitary–ovarian (HPO) axis leading to PCOS (Lewandowski et al., 2011). Hyperinsulinemia increases the GnRH pulse rate, increases ovarian androgen production, and decreases follicular maturation in the ovaries (Lewandowski et al., 2011). As a method for treating PCOS, oral contraceptives and metformin are used clinically, and oral contraceptives are used to induce sex hormone-binding globulin (SHBG) production and increase free testosterone binding, consequently reducing the symptoms of high testosterone hirsutism and inducing a return to normal menstruation (Lewandowski et al., 2011). In addition, Metformin is a commonly used treatment for type 2 diabetes to reduce insulin resistance and is used worldwide as an insulin resistance treatment for PCOS (Bauer and Cooper-Mahkorn, 2008; Kim et al., 2016; Lewandowski et al., 2011).
The use of alternative medicines by women has increased during the past 10 years; this includes the use of herbal medicines that are known to contain active pharmacological ingredients that have physiological effects on female reproduction (Whitten and Naftolin, 1998). In PCOS, medication management seems to have a moderate effect on the treatment of individual symptoms by focusing on a multidisciplinary approach (Amsterdam, 2012). Herbal medicines are complex interventions, and there is the potential for synergistic and antagonistic interactions between compounds (Williamson, 2001). Traditional alternative medicine has the advantage of active target diversity and multi pharmaceutical ingredients. Ancient documents such as the Donguibogam, and which describes principles and practices of oriental medicine, Mahuang helps diuretic effects and reduces sweating, fever, cough, asthma and metabolic dysfunction (Ahn et al., 2008; Kim et al., 2014). Mahuang-Tang (MHT) comprises six herbal medicines and has traditionally been used in the treatment of sweating, asthma, febrile disease, and obesity (Go et al., 2014; Kubo and Nishimura, 2007; Lim et al., 2016; Seo et al., 2017; Shekelle et al., 2003). However, no investigation has focused on the therapeutic effect of MHT in ovarian dysfunction disorders such as PCOS. In the present study, we investigated the effect of MHT in a rat model of PCOS by Letrozole induction. The plasma LH, FSH, estradiol, and testosterone concentrations were measured using ELISA kits. The expression of steroid receptors and steroidogenic genes was also evaluated in order to determine the potential anti-PCOS effects of MHT.
Section snippets
Preparation of MHT (Mahuang-Tang) extract
MHT comprises six herbs: Ephedrae Herba, Cinnamomi Ramulus, Glycyrrhizae Radix et Rhizoma, Armeniacae Semen, Allii Radix, and Zingiberis Rhizoma Crudus. These were obtained from a commercial vendor at Naemome Dah (Ulsan, South of Korea). The raw materials were blended at a differential ratio of 3:2:0.6:1:1:1 (w/w) (Table 1). 1612 g herbal mixtures were boiled at 100 °C for 2 h with distilled water. The extracted aqueous solution was filtered using filter paper (Whatman No. 2) and then
Quantitative analysis of major compounds in MHT extract
Confirmation of the chemical constitutions of the MHT aqueous extracts and the nine major compounds was carried out by HPLC-PDA analysis. As shown in Table 3, the linearity of the values of the correlation coefficients (r2) was obtained with the calibration curve by the nine reference compounds. We finally identified the seven of major compounds, liquiritin apioside, liquiritin, ferulic acid, coumarin, cinnamic acid, cinnamaldehyde, glycyrrhizin which was detected at 11.6, 11.8, 12.5, 16.3,
Discussion
MHT is a traditional Korean medicine consisting of six herbs: Ephedrae Herba (ephedrine), Cinnamomi Ramulus (coumarin, cinnamic acid, and cinnamaldehyde), Glycyrrhizae Radix et Rhizoma (glycyrrhizin and liquiritin), Armeniacae Semen (amygdalin), Zingiberis Rhizoma Crudus, and Allii Radix (ferulic acid) (Lim et al., 2016). The use of herbs as an alternative medicine for PCOS has been reported and is expected to prevent the onset of hormonal imbalances and improve PCOS-like symptoms in rats (
Conclusions
The present study demonstrated that MHT effectively inhibited the PCOS-like symptoms induced by Letrozole treatment. The induction of plasma LH and LH/FSH ratio level(s) in Letrozole-induced rats treated with MHT explained the ameliorating effects of this traditional prescription. Additionally, the dietary consumption of MHT regulated the mRNA level(s) of ovarian Lhr, Fshr, Pgr, Cyp11a1, and Cyp19a1 in Letrozole-induced rats at a similar level to that of the normal control. Based on these
Author contributions
H.Y., Y.H.L., S.R.L., K. P, H.W.L., and E.J.H. performed the research, analyzed the data, and wrote the manuscript; H.Y., Y.H.L., and S.R.L. performed the animal experiments and analyzed the data; H.W.L. analyzed the HPLC data and chemical profiles. All authors read and approved the final manuscript.
Declaration of competing interest
The authors declare no conflict of interest.
Acknowledgments
This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the ministry of Education (2018R1D1A1A02043102), and also supported by a grant from the Korea Institute of Oriental Medicine (grant no. K17292 and K18043).
References (32)
- et al.
Reproductive dysfunction in women with epilepsy: menstrual cycle abnormalities, fertility, and polycystic ovary syndrome
Int. Rev. Neurobiol.
(2008) - et al.
Definitions, prevalence and symptoms of polycystic ovaries and polycystic ovary syndrome
Best Pract. Res. Clin. Obstet. Gynaecol.
(2004) - et al.
Letrozole-induced polycystic ovaries in the rat: a new model for cystic ovarian disease
Arch. Med. Res.
(2004) - et al.
The anti-obesity effect of Ephedra sinica through modulation of gut microbiota in obese Korean women
J. Ethnopharmacol.
(2014) - et al.
Ovary is necessary to the health of uterus
J Gynecol Oncol
(2016) - et al.
Antipyretic effect of Mao-to, a Japanese herbal medicine, for treatment of type A influenza infection in children
Phytomedicine
(2007) Estrogen and androgen receptors: regulators of fuel homeostasis and emerging targets for diabetes and obesity
Trends Endocrinol. Metab.
(2011)- et al.
Serum antimullerian hormone levels remain stable throughout the menstrual cycle and after oral or vaginal administration of synthetic sex steroids
Fertil. Steril.
(2008) - et al.
Reproductive actions of phytoestrogens
Baillieres Clin Endocrinol Metab
(1998) Synergy and other interactions in phytomedicines
Phytomedicine
(2001)
Licorice ethanol extract improves symptoms of polycytic ovary syndrome in Letrozole-induced female rats
Integr Med Res
DONGUIBOGAM (English Version)
Consensus on women's health aspects of polycystic ovary syndrome (PCOS)
Hum. Reprod.
Species-specific differences in follicular antral sizes result from diffusion-based limitations on the thickness of the granulosa cell layer
Mol. Hum. Reprod.
What is the best treatment for women with polycystic ovarian syndrome and high LH/FSH ratio? A comparison among in vitro fertilization with GnRH agonist, GnRH antagonist and in vitro maturation
J. Endocrinol. Investig.
The effect of green tea extract on reproductive improvement in estradiol valerate-induced polycystic ovarian syndrome in rat
Iran. J. Pharm. Res. (IJPR)
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