Deer (Cervus elaphus) antler extract suppresses adhesion and migration of endometriotic cells and regulates MMP-2 and MMP-9 expression

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Abstract

Ethnopharmacological relevance

Deer antler has been used for centuries as medicine for a wide range of health problems, including various women's diseases. However, there is a safety concern related to its use in hormone-sensitive conditions, such as breast cancer and endometriosis. In the present study, we investigated the effect of deer (Cervus elaphus) antler extract (DAE) on adhesion and migration of human endometriotic cells.

Materials and methods

Adhesion, wound-healing, and transwell migration assays were performed in endometriotic cells 11Z and 12Z. Expression of matrix metalloproteinase (MMP)-2, MMP-9, TNF-α, and IL-6 were measured by real-time RT-PCR and Western blot analysis.

Results

DAE (50 and 100 μg/ml) decreased the adhesion of 11Z and 12Z cells on peritoneal mesothelial Met5-A cells. Wound-healing and transwell migration assays revealed that DAE (50 and 100 μg/ml) inhibited migration in 11Z and 12Z cells. It was further demonstrated that treatment with DAE (50 and 100 μg/ml) significantly decreased the levels of MMP-2, MMP-9, TNF-α, and IL-6.

Conclusions

These results indicate that DAE is a potential anti-endometriotic agent to inhibit the adhesion and migration of endometrial cells through the suppression of various related molecules.

Introduction

Endometriosis is a common gynaecological disorder in which functioning endometrial glands and stroma are aberrantly present in sites outside the uterine cavity. It affects 5–15% of women in the reproductive phase, and the main clinical features are infertility and chronic pelvic pain, including dysmenorrhoea. In fact, endometriosis accounts for over 20% of all cases of infertility in women (Bulun, 2009). The establishment of endometriotic lesions in the peritoneal cavity requires adhesion, migration, invasion, and proliferation of the ectopic endometrial tissue (Nisolle and Donnez, 1997, Van Langendonckt et al., 2002, Giudice and Kao, 2004). The molecular mechanism of endometriosis has been a focus of active scientific investigations over the past decade. For example, the expression levels of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) were higher in women with endometriosis than their healthy counterparts (Osteen et al., 2003). In fact, the enzymes play important roles in the ectopic adhesion, invasion and implantation, and neovascularisation of the endometrium (Chen et al., 2004, Li et al., 2006).

Various therapies have been used for treating endometriosis, including surgical and medical strategies. Current medical therapy consists mainly of hormonal suppressive therapy in which the medication causes a downregulation of the hypothalamus–pituitary–ovarian pathway. Those medical treatments are effective for relieving pain during treatment but have a high recurrence rate and significant side effects, such as hot flashes and genital atrophy (Vignali et al., 2005). Thus, novel therapeutic strategies are urgently needed to improve the clinical management of patients with endometriosis.

Deer antler (termed “Nokyong” in Korean and “Lu rong” in China) has been used in the East for centuries as a key component for prevention and/or treatment of a wide range of health problems including cardiovascular, immunological, and gynaecological disease (Huang, 1997). These traditional uses are supported by several studies showing that deer antler has anti-inflammatory effects (Takikawa et al., 1972, Zhang et al., 2000, Kim et al., 2003), anti-whiplash activities (Chen et al., 1992, Sunwoo et al., 1998, Blob and Snelgrove, 2006), and anti-aging activities (Dai et al., 2011). In Asia and Russia, deer antler has been used for a variety of gynaecological conditions, such as menstrual disorder, menopause, and infertility. However, it is generally believed that deer velvet should be avoided for women with hormone-sensitive diseases, such as endometriosis and breast cancer. To the best of our knowledge, there is no scientific evidence of the bioactivity and safety of deer antler for gynaecological diseases. In the present study, we investigate the effect of deer antler extract (DAE) on endometriosis and the associated molecular mechanism.

Section snippets

Materials

Powdered antler from Cervus elaphus Linnaeus was obtained from Kyung Hee Oriental Medical Hospital (Kyung Hee University, Seoul, South Korea) as i.p. injection grade for humans, originating from deer born and bred in New Zealand. Because deer antler has traditionally been extracted by water in oriental medicine, we used a similar extraction method using boiling water for 48 h, filtration, and lyophilisation (yield: 36.1%). The powdered extract (DAE; pH 6.8 ± 0.056) was dissolved in saline and then

The effect of DAE on endometriotic cell adhesion

Once endometrial tissue is introduced into the peritoneal cavity, it must attach, invade deep into the peritoneum covered by a mesothelial layer to establish a vascular supply, and proliferate. To investigate the effect of DAE on the cellular adhesion of endometriotic cells to the human mesothelial layer, adhesion assays using endometriotic cells (11Z and 12Z) and peritoneal mesothelial cells were performed in the presence or absence of DAE. As shown in Fig. 1, pre-incubation of endometriotic

Discussion

Current medical therapies for endometriosis, including non-steroidal anti-inflammatory agents, androgenic agents, GnRH analogues, oestrogen/progestin combined oral contraceptives, and progestins, are effective for relieving pain during treatment but do not minimise the high recurrence rate and have significant side effects (Garai et al., 2006). Thus, novel therapeutic strategies are essential to improve clinical management of patients with endometriosis.

Deer antler, traditionally used as a

Acknowledgements

This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant (to JHC) funded by the Korea Government (MEST) (No. 2009-0068979). We sincerely thank Dr. Starzinski-Powitz (Johann-Wolfgang-Goethe-Universitaet, Germany) for providing endometriotic epithelial cells as generous gifts.

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