Elsevier

Journal of Ethnopharmacology

Volume 151, Issue 3, 12 February 2014, Pages 1218-1225
Journal of Ethnopharmacology

Combination use of ferulic acid, ligustrazine and tetrahydropalmatine inhibits the growth of ectopic endometrial tissue: A multi-target therapy for endometriosis rats

https://doi.org/10.1016/j.jep.2013.12.047Get rights and content

Abstract

Ethnopharmacological relevance

Ferulic acid (FA), ligustrazine (LZ) and tetrahydropalmatine (THP) are separately isolated from Chinese Angelica, Szechwan Lovage Rhizome and Rhizoma in the Jiawei-Foshou-San formula, a popular traditional Chinese medicine for irregular menses. It has been reported that the combination use of FA+LZ+THP has similar effect on endometriosis, but the underlying mechanisms are unclear. This study was to investigate the combination effects and mechanisms of FA+LZ+THP on endometriosis rats.

Materials and methods

Fifty endometriosis rats were intragastricly treated with FA+LZ+THP for 4 wk. The volume of ectopic endometrial tissue was measured to evaluate the effects. Then the changes in hypothalamic–pituitary–ovarian axis and ERE pathway were indicated by the levels of E2, GnRH, FSH and LH, and the expressions of ER, HSP90 and COX-2, respectively. In addition, peritoneal macrophages of each rat were cultured in vitro and treated with (FA+LZ+THP)-medicated serum for 24 h. The proliferation and phagocytosis abilities, the levels of IL-1β and TNF-α, and the expression of IκBα were then measured for the changes of peritoneal macrophage activities.

Results

Combination use of FA+LZ+THP diminished the volume of the ectopic endometrial tissues (P<0.05 or P<0.01). It also decreased the E2 level, suppressed the expression of GnRH, FSH and LH, and decreased the protein expression of ER, HSP90 and COX-2 (all P<0.05 or P<0.01). The phagocytosis ability of peritoneal macrophage was enhanced by (FA+LZ+THP)-medicated serum (P<0.05) with no change of proliferation (P>0.05). Moreover, IL-1β and TNF-α were downregulated (both P<0.05 or P<0.01) and IκBα was upregulated by the (FA+LZ+THP)-medicated serum (P<0.01).

Conclusions

The combination use of FA, LZ and THP could inhibit the growth of ectopic endometrial tissue in endometriosis rats. It might be related to the down-regulation of hypothalamic–pituitary–ovarian axis, the amelioration in ERE pathway and the improvement of peritoneal macrophage activities.

Introduction

Endometriosis is a disease characterized by the presence of endometrium-like glandular tissue and stroma outside the uterus (Vinatier et al., 2001). It is thought to be a common disease with an estimated morbidity as 10–15% in women of reproductive age and most with pelvic pain and/or infertility (Eskenazi and Warner, 1997). The pathogenesis of endometriosis is still controversial. Three theories have been proposed to explain it, and of which the menstrual reflux implantation theory is mostly accepted, i.e. endometrial cells reflux to abdominal cavity following retrograde menstrual blood during menstruation (Kodati et al., 2008). Recent studies suggest that endometriosis sufferers have higher content of estrogen than normal ones (Rižner, 2009). Peritoneal macrophages of endometriosis sufferers not only grow in number, but also secrete lots of proinflammatory cytokines such as TNF-α and IL-1β. However, their phagocytic capacities were obviously weakened (Tariverdian et al., 2007). All of these abnormalities promote the development of endometriosis. Otherwise, the growth of ectopic endometrium is regulated by hormone secretion of ovarian (Chen and Li, 2005). The hypothalamus, pituitary and ovarian interacts with each other, which is essential to form a complete and coordinated neuroendocrine system, namely hypothalamic–pituitary–ovarian axis (Zhou and Xue, 2005). At present, the treatment of endometriosis is primarily by surgical eradication and/or hormone drugs, which may bring about severe adverse reaction. For example, hormone treatment can induce mood changes, body configuration changes and so on. Surgical treatment can bring about great insults to the endometriosis sufferers, and it often does not provide a definitive treatment (Evers et al., 1991). Therefore, searching for new drugs with less adverse reactions to treat endometriosis is significant.

Ferulic acid (FA), ligustrazine (LZ) and tetrahydropalmatine (THP) (the structures can be seen in Fig. 1) are three compounds isolated from Chinese Angelica, Szechwan Lovage Rhizome and Rhizoma in the Jiawei-Foshou-San formula, a popular traditional Chinese medicine for treating irregular menses. It has been reported that the combination use of FA+LZ+THP has similar effect on treating endometriosis with the Jiawei-Foshou-San formula (Wang et al., 2011, Yang et al., 2011), but the underlying mechanisms are not totally understood. Therefore, the purpose of this study is to investigate the combination effects of FA, LZ and THP on endometriosis rats' ectopic endometrial tissues and explore the relationship of this effect with the hypothalamic–pituitary–ovarian axis, the ERE pathway and the peritoneal macrophage.

Section snippets

Animals and reagents

Female Sprague–Dawley rats of 180–220 g were purchased from the Experimental Animal Center, Chongqing Medical University (Chongqing, China) and housed under controlled environment (22±2 °C, 12 h light/dark cycle, free access to food and water) in the Experimental Animal Center, College of Pharmaceutical Sciences, Southwest University (Chongqing, China). The animal approval number was SYXK 2009-0002. All the experiments were performed in accordance with China's Guidelines for Care and Use of

Combination use of FA, LZ and THP inhibits the growth of ectopic endometrial tissue

The growth of ectopic endometrial tissue in endometriosis rats is mainly evaluated by the volume change. In our study, the ectopic endometrial tissue in EMS group hardly changed in the spherical volume after treated with 0.5% CMC-Na (P>0.05). However, the volume of ectopic endometrial tissue in FA+LZ+THP 0.045 g kg−1 d−1 group had diminished by 19.60±8.60 mm3 (P<0.05). The volume of ectopic endometrial tissue in FA+LZ+THP 0.09 and 0.18 g kg−1 d−1 groups had diminished by 27.60±10.04 mm3 and 30.51±11.40 

Discussion

In traditional Chinese medicine, FA, LZ and THP were the main active constituents of Chinese Angelica, Szechwan Lovage Rhizome and Rhizoma, respectively. It was reported that all of them were related to treating for endometriosis. FA could inhibit uterine contraction (Ozaki and Ma, 1990). LZ could down-regulate the auto-secretion of RANTES and CCR5 in patients with endometriosis (Min et al., 2008). THP could suppress the growth of ectopic endometrial implants in experimentally induced

Acknowledgments

This work was supported by the Ministry of Education Doctoral Program special fund (No. 20110182110012), the Fundamental Research Funds for Central Universities(No. XDJK2010C059), the Key Projects of Chinese Medicine Research of Chongqing Municipal Health Bureau (2010[60] 2010-1-4), the Program for Innovative Research Team in University of Chongqing (2013), the Natural Science Foundation Project of CQ (CSTC, 2011jjA10010) and the Southwest University Doctor Foundation (No. SWU112102). The

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