背景:子宫腺肌病是一种常见的妇科疾病,以子宫内膜腺体和子宫肌层间质过度生长为特征,然而,其确切的病理生理学仍然不确定。新的证据表明,子宫内膜异位症和子宫腺肌病中精氨酸酶2(ARG2)的水平升高。本研究旨在确定ARG2是否参与子宫腺肌病的线粒体功能和上皮间质转化(EMT)及其潜在的潜在机制。
方法:RNA干扰抑制ARG2基因,然后进行细胞计数试剂盒(CCK-8)分析和流式细胞仪检测细胞增殖能力,细胞周期,和细胞凋亡的进展,分别。建立小鼠子宫腺肌病模型,蛋白质印迹分析,进行了线粒体膜电位(ΔΦm)检测和mPTP开放评估。
结果:沉默ARG2可有效下调子宫内膜细胞mRNA和蛋白水平的表达,导致酶活性降低和细胞活力抑制。此外,ARG2敲低诱导G0/G1细胞周期阻滞,促进细胞凋亡,并调节细胞周期和凋亡相关调节因子的表达。值得注意的是,干扰ARG2通过线粒体功能障碍诱导细胞凋亡,ROS生产,ATP耗竭,降低Bcl-2/Bax比值,释放细胞色素c,并增加Caspase-9/-3和PARP的表达。在子宫腺肌病小鼠模型中的体内研究也证明了ARG2和EMT标志物的水平升高。而siARG2治疗逆转EMT并调节炎性细胞因子。此外,发现ARG2敲低调节小鼠子宫腺肌病中的NF-κB和Wnt/β-catenin信号通路。
结论:因此,ARG2沉默可通过ROS介导的线粒体依赖性途径诱导细胞凋亡,通过抑制Ishikawa细胞中的NF-κB和Wnt/β-catenin信号通路来阻滞G0/G1细胞周期。这些发现共同表明,ARG2在子宫腺肌病的发病机理中起着至关重要的作用,并可能作为治疗干预的潜在目标。
BACKGROUND: Adenomyosis is a common gynecological disease, characterized by overgrowth of endometrial glands and stroma in the myometrium, however its exact pathophysiology still remains uncertain. Emerging evidence has demonstrated the elevated level of arginase 2 (ARG2) in endometriosis and adenomyosis. This study aimed to determine whether ARG2 involved in mitochondrial function and epithelial to mesenchymal transition (EMT) in adenomyosis and its potential underlying mechanisms.
METHODS: RNA interference was used to inhibit ARG2 gene, and then Cell Counting Kit (CCK-8) assay and flow cytometery were performed to detect the cell proliferation capacity, cell cycle, and apoptosis progression, respectively. The mouse adenomyosis model was established and RT-PCR, Western blot analysis, mitochondrial membrane potential (Δψm) detection and mPTP opening evaluation were conducted.
RESULTS: Silencing ARG2 effectively down-regulated its expression at the mRNA and protein levels in endometrial cells, leading to decreased enzyme activity and inhibition of cell viability. Additionally, ARG2 knockdown induced G0/G1 cell cycle arrest, promoted apoptosis, and modulated the expression of cell cycle- and apoptosis-related regulators. Notably, the interference with ARG2 induces apoptosis by mitochondrial dysfunction, ROS production, ATP depletion, decreasing the Bcl-2/Bax ratio, releasing Cytochrome c, and increasing the expression of Caspase-9/-3 and PARP. In vivo study in a mouse model of adenomyosis demonstrated also elevated levels of ARG2 and EMT markers, while siARG2 treatment reversed EMT and modulated inflammatory cytokines. Furthermore, ARG2 knockdown was found to modulate the NF-κB and Wnt/β-catenin signaling pathways in mouse adenomyosis.
CONCLUSIONS: Consequently, ARG2 silencing could induce apoptosis through a mitochondria-dependent pathway mediated by ROS, and G0/G1 cell cycle arrest via suppressing NF-κB and Wnt/β-catenin signaling pathways in Ishikawa cells. These findings collectively suggest that ARG2 plays a crucial role in the pathogenesis of adenomyosis and may serve as a potential target for therapeutic intervention.