METHODS: The infarcted areas and cardiac function were evaluated, and the expression level of ZC3H13 was measured in AMI rats that were induced by isoproterenol. Meanwhile, oxygen glucose deprivation (OGD) in vitro model was induced to investigate the alterations on inflammation, oxidative stress and ferroptosis. The m6A modification site of lncRNA93358 modified by ZC3H13 was predicted using bioinformatics, and the interaction between ZC3H13 and lncRNA93358 was verified using the dual-luciferase reporter assays. ZC3H13 was overexpressed and lncRNA93358 was silenced to study their regulatory role in cell death, inflammation, oxidative stress and ferroptosis in AMI.
RESULTS: Significant decreased expression of ZC3H13 was observed in AMI rats, with impaired cardiac function, enhanced inflammation and oxidative stress. ZC3H13 targeted the modification site GGACC of lncRNA93358 and downregulated lncRNA93358. Silencing lncRNA93358 inhibited cell death, reduced the levels of inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β, suppressed oxidative stress-related indicators (lactate dehydrogenase (LDH), reactive oxygen species (ROS), glutathione (GSH) and malondialdehyde (MDA), as well as downregulated ferroptosis-related acyl-CoA synthetase long chain family member 4 (ACSL4), prostaglandin-endoperoxide synthase 2 (PTGS2) and glutathione peroxidase 4 (GPX4). The effect of silencing lncRNA93358 was further enhanced by overexpression of ZC3H13.
CONCLUSIONS: This study reveals the ZC3H13-mediated epigenetic RNA modification targeting lncRNA93358 and suggests that ZC3H13 overexpression may be a promising approach for AMI treatment.
方法:评估梗死面积和心功能,在异丙肾上腺素诱导的AMI大鼠中测定ZC3H13的表达水平。同时,氧葡萄糖剥夺(OGD)体外模型的诱导,研究炎症的变化,氧化应激和铁死亡。用生物信息学方法预测ZC3H13修饰的lncRNA93358的m6A修饰位点,ZC3H13和lncRNA93358之间的相互作用使用双荧光素酶报告基因测定进行验证。ZC3H13被过表达,lncRNA93358被沉默以研究它们在细胞死亡中的调节作用,炎症,AMI中的氧化应激和铁凋亡。
结果:在AMI大鼠中观察到ZC3H13的表达显著降低,心脏功能受损,增强的炎症和氧化应激。ZC3H13靶向lncRNA93358的修饰位点GGACC并下调lncRNA93358。沉默lncRNA93358抑制细胞死亡,降低炎症细胞因子肿瘤坏死因子(TNF)-α的水平,白细胞介素(IL)-6和IL-1β,抑制氧化应激相关指标(乳酸脱氢酶(LDH),活性氧(ROS),谷胱甘肽(GSH)和丙二醛(MDA),以及下调的铁凋亡相关的酰基辅酶A合成酶长链家族成员4(ACSL4),前列腺素-内过氧化物合酶2(PTGS2)和谷胱甘肽过氧化物酶4(GPX4)。沉默IncRNA93358的作用通过ZC3H13的过表达进一步增强。
结论:这项研究揭示了ZC3H13介导的靶向lncRNA93358的表观遗传RNA修饰,并表明ZC3H13过表达可能是AMI治疗的有希望的方法。