Role of cathepsin E in decidual macrophage of patients with recurrent miscarriage
Role of cathepsin E in decidual macrophage of patients with recurrent miscarriage
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DOI：
10.1093/molehr/gau008
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发表时间：
2014-05-01
影响因子：
4
通讯作者：
Sugiura-Ogasawara, Mayumi
中科院分区：
文献类型：
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作者：
Goto, Shinobu;Ozaki, Yasuhiko;Sugiura-Ogasawara, Mayumi
在以前的研究中，我们报道了组织蛋白酶-半胱氨酸蛋白酶抑制剂系统导致早期妊娠子宫内膜功能障碍。在这里，我们调查了组织蛋白酶E的存在和贡献，在早孕复发性流产（RM）患者。还在CatE（-/-）小鼠中分析了组织蛋白酶缺乏对生育力和雌性生殖器官的影响。在知情同意的情况下，在医院环境中进行人体研究。收集RM患者孕早期（4-6孕周，n = 21）宫颈粘液，并前瞻性比较妊娠结局。采用SDS-PAGE和westernblot方法检测49例RM患者和24例正常妊娠行选择性人工流产患者蜕膜组织中组织蛋白酶E的表达。从RM患者（n = 6）分离蜕膜巨噬细胞，并通过脂多糖（LPS）和干扰素γ（IFN-γ）刺激。小鼠模型的结果显示，CatE（-/-）小鼠具有生育能力，但窝仔数显著较少。CatE（-/-）小鼠子宫出现肉芽组织。在人类样本中，与没有RM的患者相比，流产患者宫颈粘液中组织蛋白酶E的蛋白酶活性（用荧光猝灭底物（KYS-1）测定）显著降低。组织蛋白酶E在RM患者蜕膜中的表达，通过SDS-PAGE半定量，western blot分析，与无RM患者相比，RM患者的表达显著降低。组织蛋白酶E免疫荧光双染法显示，所有蜕膜组织中均可见CD 14或CD 68阳性细胞。经LPS和IFN-γ刺激后，RM患者蜕膜巨噬细胞裂解液中组织蛋白酶E的表达明显低于对照组。结果提示，蜕膜巨噬细胞分泌的组织蛋白酶E活性降低可能是部分RM患者流产的原因。
In a previous study, we reported that the cathepsin-cystatin system caused endometrial dysfunction in early pregnancy. Here, we investigated the existence and contribution of cathepsin E in early pregnancy in patients with recurrent miscarriage (RM). The effect of cathepsin deficiency on fertility and female reproductive organs were also analyzed in CatE(-/-) mice. Human studies were conducted in a hospital setting, with informed consent. Cervical mucus was collected from RM patients in early pregnancy (4-6 gestational weeks, n = 21), and the pregnancy outcome was compared prospectively. The cathepsin E expression in decidua of RM patients (n = 49) and normal pregnant women undergoing elective surgical abortion (n = 24) was measured using SDS-PAGE, and western blot analysis. Decidual macrophages were isolated from RM patients (n = 6) and stimulated by lipopolysaccharide (LPS) and interferon gamma (IFN-gamma). Results from the mouse model showed that CatE(-/-) mice were fertile, but the litter number was significantly smaller. The uterus of CatE(-/-) mice showed granulation tissue. In human samples, protease activity of cathepsin E measured with Fluorescence-Quenching Substrate (KYS-1) in cervical mucus of patients who developed miscarriage was markedly decreased compared with patients without RM. The expression of cathepsin E in decidua, semi-quantified by SDS-PAGE, western blot analysis was significantly lower in RM patients compared with patients without RM. By double staining immunofluorescence, the staining of cathepsin E was observed in CD14 or CD68 positive cells in all deciduas. Upon stimulation with LPS and IFN-gamma, the expression of cathepsin E in cell lysate of decidual macrophages was markedly reduced in RM patients compared with controls. The results suggested that decreased activity of cathepsin E produced by decidual macrophages might be responsible for the induction of miscarriages in some RM patients.