UNASSIGNED: Human lens epithelial cells (SRA01/04 cells) and rat transparent lens were cultured with H2O2 to establish cell oxidative stress models and rat cataract models. Immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot assays were employed to detect Slit2 levels within age-related cataracts(ARC) lens anterior capsule samples, rat cataract models, and cell oxidative stress models. In this study, qRT-PCR and Western blot assays were performed to derermine E-cadherin, N-cadherin, occludens1(ZO-1), α-SMA(α‑smooth muscle actin), Bcl-2, Bax, p-AKT, and AKT levels. In addition, Flow cytometry were performed to examine reactive oxygen species (ROS) and cell apoptosis. Cell viability, invasion, and migration were detected by CCK8, Transwell, and Wound healing.
UNASSIGNED: Increased expression of Slit2 was found in ARC lens anterior capsule samples, H2O2-induced rat cataract models, and Human lens epithelial cells (HLECs) oxidative stress models. H2O2 significantly increased cell apoptosis and ROS generation, also accelerating cell migration, invasion, and epithelial-mesenchymal transition (EMT). In addition, H2O2 treatment repressed AKT phosphorylation and cell viability. Knock-down of Slit2 promoted cell viability and AKT phosphorylation levels, as well as repressed cell invasion, migration, apoptosis, ROS production and EMT.
UNASSIGNED: Slit2 promoted lens epithelial cells oxidative stress damage via the AKT signalling pathways, providing a novel insight in ARC treatment.
■用H2O2培养人晶状体上皮细胞(SRA01/04细胞)和大鼠透明晶状体,建立细胞氧化应激模型和大鼠白内障模型。免疫组织化学,定量实时聚合酶链反应(qRT-PCR),和蛋白质印迹测定法用于检测年龄相关性白内障(ARC)晶状体前囊样品中的Slit2水平,大鼠白内障模型,和细胞氧化应激模型。在这项研究中,进行qRT-PCR和Western印迹分析,N-钙黏着蛋白,闭塞1(ZO-1),α-SMA(α-平滑肌肌动蛋白),Bcl-2,Bax,p-AKT,AKT水平。此外,进行流式细胞术以检查活性氧(ROS)和细胞凋亡。细胞活力,入侵,迁移被CCK8、Transwell、伤口愈合。
■在ARC晶状体前囊样本中发现Slit2的表达增加,H2O2诱导的大鼠白内障模型,和人晶状体上皮细胞(HLECs)氧化应激模型。H2O2显著增长细胞凋亡和ROS生成,也加速细胞迁移,入侵,和上皮-间质转化(EMT)。此外,H2O2处理抑制AKT磷酸化和细胞活力。敲低Slit2促进细胞活力和AKT磷酸化水平,以及抑制细胞入侵,迁移,凋亡,ROS生产和EMT。
■Slit2通过AKT信号通路促进晶状体上皮细胞氧化应激损伤,在ARC治疗提供了一个新的见解。