Puerarin Suppresses Invasion and Vascularization of Endometriosis Tissue Stimulated by 17β-Estradiol
et al. (2011) Puerarin Suppresses Invasion and Vascularization of Endometriosis Tissue Stimulated by 17b-Estradiol. PLoS
ONE 6(9): e25011. doi:10.1371/journal.pone.0025011
Puerarin Suppresses Invasion and Vascularization of Endometriosis Tissue Stimulated by 17b-Estradiol
Dan Wang 0
Yuhuan Liu 0
Jie Han 0
Dongxia Zai 0
Mei Ji 0
Wen Cheng 0
Ling Xu 0
Luxi Yang 0
Miaoxia He 0
Jian Ni 0
Zailong Cai 0
Chaoqin Yu 0
Jean-Marc A. Lobaccaro, Clermont Universite, France
0 1 Department of Chinese Traditional Medicine, Changhai Hospital, Second Military Medical University , Shanghai , China , 2 Clinical Research Center, Changhai Hospital, Second Military Medical University , Shanghai , China , 3 Department of Obstetrics and Gynecology, Changhai Hospital, Second Military Medical University , Shanghai , China , 4 Department of Pathology, Changhai Hospital, Second Military Medical University , Shanghai , China , 5 Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University , Shanghai , China
Background: Puerarin, a phytoestrogen with a weak estrogenic effect, binds to estrogen receptors, thereby competing with 17b-estradiol (E2) and producing an anti-estrogenic effect. This study was to investigate whether puerarin could suppress the invasion and vascularization of E2-stimulated endometriotic tissue. Methodology/Principal Findings: The endometriotic stromal cells (ESCs) were successfully established and their invasive ability under different treatments was assessed through a Transwell Assay. Simultaneously, matrix metallopeptidase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) were detected by western blotting. Vascularization of endometriotic tissues was observed by chicken chorioallantoic membrane (CAM) assay. The staining of MMP-9, intercellular adhesion molecule 1 (ICAM-1), TIMP-1, and vascular endothelial growth factor (VEGF) in grafted endometriotic tissues was examined using immunohistochemistry analysis. The purity of ESCs in isolated cells was .95%, as determined by the fluoroimmunoassay of vimentin. E2 (1028 mol/L) promoted the invasiveness of ESCs by increasing MMP-9 accumulation and decreasing TIMP-1 accumulation. Interestingly, puerarin (1029 mol/L) significantly reversed these effects (P,0.01). The CAM assay indicated that puerarin (1029 mol/L) also inhibited the angiopoiesis of endometriotic tissue stimulated by the E2 (1028 mol/L) treatment (P,0.05). Accordingly, immunohistochemistry showed that the accumulation of MMP-9, ICAM-1, and VEGF was reduced whereas that of TIMP-1 increased in the combination treatment group compared with the E2 treatment group. Conclusions/Significance: This study demonstrated that puerarin could suppress the tissue invasion by ESCs and the vascularization of ectopic endometrial tissues stimulated by E2, suggesting that puerarin may be a potential drug for the treatment of endometriosis.
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. These authors contributed equally to this work.
Endometriosis, a common, benign, estrogen-dependent disease
affecting 3%10% of women of reproductive age, is defined as
the presence of endometrial glands and stroma outside of the
uterine cavity [1]. It is not a malignant disorder but it has
similarities to a developing tumor. For instance, endometriotic
cells can be both locally and distantly metastatic, that is they can
attach to, invade and damage other tissues [2,3]. The initial
phase of endometriosis is an invasive event that requires
extracellular matrix (ECM) breakdown. The key enzymes that
regulate the ECM are the matrix metalloproteinases (MMPs), and
peritoneal invasion by endometrial tissue is thought to be
dependent on MMPs and their corresponding tissue inhibitors
of metalloproteinases (TIMPs). MMPs play a pivotal role in the
cyclic changes of growth and tissue breakdown that occur in the
endometrium. MMPs are usually synthesized during the
proliferative phase and are stimulated by estrogen [4]. Several studies
have shown an increase in the accumulation of MMP-1, MMP-2,
MMP-3, MMP-7, and MMP-9 in endometriotic tissues.
Alteration of MMP-9 and MMP-2 is an important factor in the
development of endometriosis [2,3].
Vascularization of endometriotic implants is probably one of the
most important factors in the invasion of other tissues by
endometrial cells. Angiogenesis is controlled by a number of
inducers; of particular importance is the vascular endothelial
growth factor (VEGF) family, which is significant in processes
characterized by both physiological and pathological angiogenesis.
Accumulation of the VEGF gene in normal human endometrial
cells is acutely upregulated by E2 in vitro [5]. VEGF
immunostaining has been observed in the epithelium of endometriotic
implants, particularly in hemorrhagic red implants [6].
Most of the current medical treatments for endometriosis aim to
downregulate estrogen activity. Historically, medical therapies
have included contraceptive steroids, progestogens, and agonists of
gonadotropin-releasing hormone (Gn (...truncated)