Cao Yang, Ye Qing, Zhuang Mengfei, Xie Shuwu, Zhong Ruihua, Cui Jingang, Zhou Jieyun, Zhu Yan, Zhang Tingting, Cao Lin
Department of Gynecology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Department of Neurology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
PLoS One. 2017 Nov 15;12(11):e0186520. doi: 10.1371/journal.pone.0186520. eCollection 2017.
This study aimed to investigate the link between the inhibitory effect of ginsenoside Rg3 on the ectopic endometrium growth and the VEGFR-2-mediated PI3K/Akt/mTOR signaling pathway, a mechanism known to inhibit angiogenesis and induce ectopic endometrial cell apoptosis.
A model of endometriosis was established by allotransplantation in rats. The rats were randomly divided into 5 groups: the ginsenoside Rg3 low-dose group (group A,5mg/kgBW/d of ginsenoside Rg3), the ginsenoside Rg3 high-dose group (group B, 10mg/kgBW/d of ginsenoside Rg3), the gestrinone group (group C, 0.5mg/kgBW/d of gestrinone), the control group (groupD, 10ml/kg BW/d of 0.5%CMC-Na) and the ovariectomized group (group E, 10ml/kgBW/d of 0.5%CMC-Na). Rats were executed after 21 days of continuous administration. The ectopic endometrium volume was measured and the inhibitory rate was calculated. The levels of serum estradiol (E2) and progesterone (P) were detected by Electro-Chemiluminescence Immunoassay (ECLI). The protein expressionof VEGF, VEGFR-2, p-Akt, and p-mTOR inthe ectopic endometrium wastested by immunohistochemistry(IHC) and Western Blotting. The mRNA expression levels of VEGF, VEGFR-2, Akt, and mTOR were tested by Real-Time Polymerase Chain Reaction (PCR). The apoptosis rate of the ectopic endometrial cells was detected by Terminal Deoxynucleotidyl Transferase-mediated Digoxigenin-dUTP Nick-End Labeling Assay(TUNEL).
Tissue measurements revealed a dose-dependent inhibition effect of ginsenoside Rg3 on the growth of the ectopic endometrium in treated rats compared to controls. Immunohistochemical and Western Blotting assays confirmed that the expression of VEGF, p-Akt, and p-mTOR was down-regulated in ginsenoside Rg3 -treated lesions. Real-time PCR results also showed that the mRNA expression levels of VEGF, Akt, and mTOR in the ectopic endometrium were reduced.
The present study demonstrates, for the first time, that ginsenoside Rg3 suppresses angiogenesis in developing endometrial lesions. The ginsenoside Rg3 inhibitory effect on the growth of the ectopic endometrium in EMs rats might occur through the blocking of the VEGFR-2-mediated PI3K/Akt/mTOR signaling pathway, thus halting angiogenesis and promoting the apoptosis of ectopic endometrial cells.
本研究旨在探讨人参皂苷Rg3对异位子宫内膜生长的抑制作用与VEGFR-2介导的PI3K/Akt/mTOR信号通路之间的联系，该信号通路是一种已知的抑制血管生成并诱导异位子宫内膜细胞凋亡的机制。
通过同种异体移植在大鼠中建立子宫内膜异位症模型。将大鼠随机分为5组：人参皂苷Rg3低剂量组（A组，人参皂苷Rg3 5mg/kg体重/天）、人参皂苷Rg3高剂量组（B组，人参皂苷Rg3 10mg/kg体重/天）、孕三烯酮组（C组，孕三烯酮0.5mg/kg体重/天）、对照组（D组，0.5%羧甲基纤维素钠10ml/kg体重/天）和去卵巢组（E组，0.5%羧甲基纤维素钠10ml/kg体重/天）。连续给药21天后处死大鼠。测量异位子宫内膜体积并计算抑制率。采用电化学发光免疫分析法（ECLI）检测血清雌二醇（E2）和孕酮（P）水平。通过免疫组织化学（IHC）和蛋白质印迹法检测异位子宫内膜中VEGF、VEGFR-2、p-Akt和p-mTOR的蛋白表达。采用实时聚合酶链反应（PCR）检测VEGF、VEGFR-2、Akt和mTOR的mRNA表达水平。采用脱氧核糖核苷酸末端转移酶介导的地高辛-dUTP缺口末端标记法（TUNEL）检测异位子宫内膜细胞的凋亡率。
组织测量显示，与人参皂苷Rg3处理组相比，人参皂苷Rg3对治疗大鼠异位子宫内膜的生长具有剂量依赖性抑制作用。免疫组织化学和蛋白质印迹分析证实，在人参皂苷Rg3处理的病变中，VEGF、p-Akt和p-mTOR的表达下调。实时PCR结果还显示，异位子宫内膜中VEGF、Akt和mTOR的mRNA表达水平降低。
本研究首次证明人参皂苷Rg3可抑制子宫内膜病变发展过程中的血管生成。人参皂苷Rg3对子宫内膜异位症大鼠异位子宫内膜生长的抑制作用可能是通过阻断VEGFR-2介导的PI3K/Akt/mTOR信号通路，从而阻止血管生成并促进异位子宫内膜细胞凋亡。
