Decreased nitric oxide content mediated by asymmetrical dimethylarginine and protein L-arginine methyltransferase 3 in macrophages induces trophoblast apoptosis: a potential cause of recurrent miscarriage
Decreased nitric oxide content mediated by asymmetrical dimethylarginine and protein L-arginine methyltransferase 3 in macrophages induces trophoblast apoptosis: a potential cause of recurrent miscarriage
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巨噬细胞中不对称二甲基精氨酸和蛋白质 L-精氨酸甲基转移酶 3 介导的一氧化氮含量减少会诱导滋养层细胞凋亡：这是反复流产的潜在原因。
DOI：
10.1093/humrep/deab225
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发表时间：
2021-10-13
影响因子：
6.1
通讯作者：
Jin, Li-Ping
中科院分区：
文献类型：
--
作者：
Hao, Fan;Tang, Lin-Chen;Jin, Li-Ping
研究问题：蛋白质L-精氨酸甲基转移酶3（PRMT 3）/不对称二甲基精氨酸（ADMA）/一氧化氮（NO）通路是否参与复发性流产（RM）的发生，其潜在机制是什么？总结回答：PRMT 3和ADMA水平升高抑制蜕膜中NO的形成，从而损害母胎界面滋养层细胞的功能。已知：NO生物利用度降低与RM相关。ADMA，一氧化氮合酶（NOS）的内源性抑制剂，是来自于甲基化的蛋白质精氨酸残基的PRMTs和作为一个预测死亡率在危重病。研究设计，规模，持续时间：共145名妇女RM和149名健康妇女进行选择性终止早期正常妊娠。CBA/J雌性小鼠96只，DBA/2雄性小鼠24只，BALB/c雄性小鼠24只。CBA/J × DBA/2交配组为流产组，CBA/J × BALB/c交配组为正常对照组。然后将CBA/J妊娠小鼠分为四组：（i）正常+媒介物组（n=28），（ii）流产+媒介物组（n=28），（iii）正常+SGC 707（PRMT 3抑制剂）组（n=20）和（iv）流产+SGC 707组（n=20）。所有注射均在妊娠第0.5、3.5和6.5天腹膜内进行。在妊娠第8.5、9.5和10.5天采集蜕膜组织。对象/材料、地点、方法：采用常规检测试剂盒和蛋白质印迹法检测蜕膜组织中NO浓度、ADMA含量、NOS活性、NOS和PRMTs表达水平。在蜕膜基质细胞、巨噬细胞和自然杀伤细胞中进一步分析PRMT 3表达。建立了蜕膜巨噬细胞（DMs）与HTR-8/SVneo滋养层细胞共培养体系，研究PRMT 3/ADMA/NO信号通路的作用。通过Annexin V-异硫氰酸荧光素/碘化丙啶染色分析滋养层细胞凋亡。本研究采用CBA/J × DBA/2小鼠模型，观察了SGC 707对RM小鼠胚胎吸收率的影响。主要结果和机会作用：结果显示：RM患者蜕膜组织中NO含量和NOS活性降低，ADMA含量和PRMT 3表达增加。此外，与正常对照组相比，PRMT 3在RM患者的巨噬细胞中表达显著上调，但在自然杀伤细胞或蜕膜基质细胞中不表达。PRMT 3的抑制导致巨噬细胞中ADMA积累的显著减少和NO浓度的增加。当与经SGC 707和ADMA处理的DM共培养时，滋养层细胞凋亡分别被抑制和诱导。体内实验表明，给予SGC 707可降低CBA/J x DBA/2小鼠的胚胎吸收率。主要原因是，每个组织需要保留用于临床诊断，并且每个组织中只有一小块可以被切割和收集用于本研究。更广泛的意义的发现：我们的研究结果表明，PRMT 3/ADMA/NO通路是一个潜在的标记物和靶点，用于RM的临床诊断和治疗。
STUDY QUESTION: Is the protein l-arginine methyltransferase 3 (PRMT3)/asymmetrical dimethylarginine (ADMA)/nitric oxide (NO) pathway involved in the development of recurrent miscarriage (RM), and what is the potential mechanism?SUMMARY ANSWER: Elevated levels of PRMT3 and ADMA inhibit NO formation in the decidua, thereby impairing the functions of trophoblast cells at the maternal-foetal interface.WHAT IS KNOWN ALREADY: Decreased NO bioavailability is associated with RM. ADMA, an endogenous inhibitor of nitric oxide synthase (NOS), is derived from the methylation of protein arginine residues by PRMTs and serves as a predictor of mortality in critical illness.STUDY DESIGN, SIZE, DURATION: A total of 145 women with RM and 149 healthy women undergoing elective termination of an early normal pregnancy were enrolled. Ninety-six female CBA/J, 24 male DBA/2 and 24 male BALB/c mice were included. CBA/J x DBA/2 matings represent the abortion group, while CBA/J x BALB/c matings represent the normal control group. The CBA/J pregnant mice were then categorised into four groups: (i) normal + vehicle group (n=28), (ii) abortion + vehicle group (n=28), (iii) normal + SGC707 (a PRMT3 inhibitor) group (n=20) and (iv) abortion + SGC707 group (n=20). All injections were made intraperitoneally on Days 0.5, 3.5 and 6.5 of pregnancy. Decidual tissues were collected on Days 8.5, 9.5 and 10.5 of gestation. The embryo resorption rates were calculated on Day 9.5 and Day 10.5 of gestation.PARTICIPANTS/MATERIALS, SETTING, METHODS: NO concentration, ADMA content, NOS activity, expression levels of NOS and PRMTs in decidual tissues were determined using conventional assay kits or western blotting. PRMT3 expression was further analysed in decidual stromal cells, macrophages and natural killer cells. A co-culture system between decidual macrophages (DMs) and HTR-8/SVneo trophoblasts was constructed to study the roles of the PRMT3/ADMA/NO signalling pathway. Trophoblast apoptosis was analysed via Annexin V-fluorescein isothiocyanate/propidium iodide staining. CBA/J x DBA/2 mouse models were used to investigate the effects of SGC707 on embryo resorption rates.MAIN RESULTS AND THE ROLE OF CHANCE: Our results show that NO concentration and NOS activity were decreased, but ADMA content and PRMT3 expression were increased in the decidua of RM patients. Moreover, compared with the normal control subjects, PRMT3 expression was significantly up-regulated in the macrophages but not in the natural killer cells or stromal cells of the decidua from RM patients. The inhibition of PRMT3 results in a significant decrease in ADMA accumulation and an increase in NO concentration in macrophages. When co-cultured with DMs, which were treated with SGC707 and ADMA, trophoblast apoptosis was suppressed and induced, respectively. In vivo experiments revealed that the administration of SGC707 reduced the embryo resorption rate of CBA/J x DBA/2 mice.LIMITATIONS, REASONS FOR CAUTION: All sets of experiments were not performed with the same samples. The main reason is that each tissue needs to be reserved for clinical diagnosis and only a small piece of each tissue can be cut and collected for this study.WIDER IMPLICATIONS OF THE FINDINGS: Our results indicate that the PRMT3/ADMA/NO pathway is a potential marker and target for the clinical diagnosis and therapy of RM.