Yoshino Osamu, Ono Yosuke, Honda Masako, Hattori Kyoko, Sato Erina, Hiraoka Takehiro, Ito Masami, Kobayashi Mutsumi, Arai Kenta, Katayama Hidekazu, Tsuchida Hiroyoshi, Yamada-Nomoto Kaori, Iwahata Shunsuke, Fukushi Yoshiyuki, Wada Shinichiro, Iwase Haruko, Koga Kaori, Osuga Yutaka, Iwaoka Michio, Unno Nobuya
Department of Obstetrics and Gynecology, Kitasato University School of Medicine, Kanagawa 252-0375, Japan.
Department of Obstetrics and Gynecology, Teine Keijinkai Hospital, Hokkaido 006-0811, Japan.
Biomedicines. 2020 Oct 31;8(11):467. doi: 10.3390/biomedicines8110467.
Relaxin (RLX)-2, produced by the corpus luteum and placenta, is known to be potentially effective in fibrotic diseases of the heart, lungs, kidneys, and bladder; however, its effectiveness in endometriosis has not yet been investigated. In the present study, we conducted a comprehensive study on the effect of RLX-2 on endometriosis. We checked the expressions of LGR-7, a primary receptor of RLX-2, in endometriomas using immunohistochemistry. Endometriotic stromal cells (ESCs) purified from surgical specimens were used in in vitro experiments. The effects of RLX-2 on ESCs were evaluated by quantitative-PCR, ELISA, and Western blotting. Gel contraction assay was used to assess the contraction suppressive effect of RLX-2. The effect of RLX-2 was also examined in the endometriosis mouse model. LGR-7 was expressed in endometriotic lesions. In ESCs, RLX-2 increased the production of cAMP and suppressed the secretion of interleukin-8, an inflammatory cytokine, by 15% and mRNA expression of fibrosis-related molecules, plasminogen activator inhibitor-1 (PAI-1), and collagen-I by approximately 50% ( < 0.05). In the gel contraction assay, RLX-2 significantly suppressed the contraction of ESCs, which was cancelled by removing RLX-2 from the medium or by adding H89, a Protein Kinase A (PKA) inhibitor. In ESCs stimulated with RLX-2, p38 MAPK phosphorylation was significantly suppressed. In the endometriosis mouse model, administration of RLX-2 significantly decreased the area of the endometriotic-like lesion with decreasing fibrotic component compared to non-treated control ( = 0.01). RLX-2 may contribute to the control of endometriotic lesion by suppressing fibrosis, scar formation, and inflammation.
松弛素(RLX)-2由黄体和胎盘产生,已知其在心脏、肺、肾脏和膀胱的纤维化疾病中可能有效;然而,其在子宫内膜异位症中的有效性尚未得到研究。在本研究中,我们对RLX-2对子宫内膜异位症的影响进行了全面研究。我们使用免疫组织化学检查了RLX-2的主要受体LGR-7在子宫内膜瘤中的表达。从手术标本中纯化的子宫内膜异位症基质细胞(ESC)用于体外实验。通过定量PCR、酶联免疫吸附测定(ELISA)和蛋白质免疫印迹法评估RLX-2对ESC的影响。凝胶收缩试验用于评估RLX-2的收缩抑制作用。还在子宫内膜异位症小鼠模型中研究了RLX-2的作用。LGR-7在子宫内膜异位症病变中表达。在ESC中,RLX-2增加了环磷酸腺苷(cAMP)的产生,并使炎症细胞因子白细胞介素-8的分泌减少了15%,使纤维化相关分子纤溶酶原激活物抑制剂-1(PAI-1)和I型胶原蛋白的信使核糖核酸(mRNA)表达减少了约50%(P<0.05)。在凝胶收缩试验中,RLX-2显著抑制了ESC的收缩,通过从培养基中去除RLX-2或添加蛋白激酶A(PKA)抑制剂H89可消除这种抑制作用。在用RLX-2刺激的ESC中,p38丝裂原活化蛋白激酶(MAPK)的磷酸化显著受到抑制。在子宫内膜异位症小鼠模型中,与未治疗的对照组相比,给予RLX-2显著减小了子宫内膜样病变的面积,纤维化成分减少(P=0.01)。RLX-2可能通过抑制纤维化、瘢痕形成和炎症来控制子宫内膜异位症病变。