Xie Yazhen, Lu Qibin
Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China.
Taicang TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Taicang, Jiangsu, China.
Evid Based Complement Alternat Med. 2020 Dec 14;2020:1967698. doi: 10.1155/2020/1967698. eCollection 2020.
Anzi Heji (AZHJ) has been used to treat anticardiolipin antibody- (ACA-) positive pregnant women at risk of spontaneous abortion for many years. The aim of this study was to investigate the protective mechanism of AZHJ in a mouse model of ACA-positive pregnancy at risk of spontaneous abortion using label-free quantitative proteomics. Mice were divided into three groups: normal pregnant mice (control group), ACA-positive pregnant mice administered normal saline (model group), and ACA-positive pregnant mice administered AZHJ (AZHJ group). The model was established by injecting 2-glycoprotein I (GPI) into mice for 18 days. The DEPs and their functions were analyzed by label-free quantitative proteomic and bioinformatic analyses. The levels of IL-6, IL-10, ACA, and TNF- in the serum and placentas of the mice were measured by enzyme-linked immunosorbent assays (ELISAs). Proteomic data were validated by western blot analysis. The abnormal serum and placental levels of IL-6, ACA, and TNF- in the model group were reversed by AZHJ. There were 39 upregulated and 10 downregulated DEPs in the AZHJ group relative to the model group. Bioinformatic analysis revealed that the DEPs were mainly involved in nucleic acid binding, signal conduction, and posttranslational modification. The placental levels of T-cell immunoglobulin mucin 3 (Tim-3) and Toll-like receptor 4 (TLR4) expression and AKT phosphorylation in the three groups were consistent with the proteomic findings. Tim-3/AKT signaling is involved in maternal-fetal immune tolerance, while TLR4 is associated with inflammatory responses. Collectively, these results indicate that AZHJ may exert its protective effect in ACA-positive pregnant mice by regulating the maternal-fetal immune tolerance and inflammatory response.
安胎合剂（AZHJ）多年来一直用于治疗有自然流产风险的抗心磷脂抗体（ACA）阳性孕妇。本研究旨在使用无标记定量蛋白质组学研究AZHJ在有自然流产风险的ACA阳性妊娠小鼠模型中的保护机制。将小鼠分为三组：正常妊娠小鼠（对照组）、给予生理盐水的ACA阳性妊娠小鼠（模型组）和给予AZHJ的ACA阳性妊娠小鼠（AZHJ组）。通过给小鼠注射β2糖蛋白I（GPI）18天建立模型。通过无标记定量蛋白质组学和生物信息学分析对差异表达蛋白（DEPs）及其功能进行分析。通过酶联免疫吸附测定（ELISA）测量小鼠血清和胎盘中白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、ACA和肿瘤坏死因子-α（TNF-α）的水平。通过蛋白质印迹分析验证蛋白质组学数据。AZHJ逆转了模型组中异常的血清和胎盘IL-6、ACA和TNF-α水平。与模型组相比，AZHJ组有39个上调的DEPs和10个下调的DEPs。生物信息学分析显示，DEPs主要参与核酸结合、信号传导和翻译后修饰。三组中T细胞免疫球蛋白粘蛋白3（Tim-3）和Toll样受体4（TLR4）的胎盘表达水平以及AKT磷酸化与蛋白质组学结果一致。Tim-3/AKT信号传导参与母胎免疫耐受，而TLR4与炎症反应相关。总体而言，这些结果表明AZHJ可能通过调节母胎免疫耐受和炎症反应在ACA阳性妊娠小鼠中发挥保护作用。
