Abstract：
Heart failure (HF) is a complex clinical syndrome associated with significant morbidity and mortality. Dysregulation of long non-coding RNA (lncRNA) has been implicated in the pathogenesis of HF. The present study aims to investigate the role of lncRNA HOX transcript antisense RNA (HOTAIR) in cardiomyocyte pyroptosis in a murine HF model. A murine HF model was established through transverse aortic contraction surgery, and an in vitro HF cell model was developed by treating HL-1 cells with H2O2. HOTAIR was overexpressed in TAC mice and HL-1 cells via pcDNA3.1-HOTAIR transfection. Cardiac function was assessed in TAC mice, and myocardial changes were evaluated using HE staining. The expression of NLRP3 was examined by immunohistochemistry. Myocardial injury markers and pyroptosis-related inflammatory cytokines were quantified using ELISA. Protein levels of NLRP3, cleaved-caspase-1, and GSDMD-N were analyzed by Western blot. Dual-luciferase assays and RNA immunoprecipitation were employed to confirm the binding interactions between HOTAIR and miR-17-5p, miR-17-5p and RORA. Functional rescue experiments were conducted by overexpressing miR-17-5p or silencing RORA in HL-1 cells. HOTAIR exhibited reduced expression in TAC mice and H2O2-induced cardiomyocytes. Overexpression of HOTAIR ameliorated cardiac dysfunction, reduced myocardial pathological injury, enhanced cardiomyocyte viability, and decreased myocardial injury and pyroptosis. HOTAIR interacted with miR-17-5p to repress RORA transcription. Overexpression of miR-17-5p or silencing of RORA abolished the inhibitory effect of HOTAIR overexpression on cardiomyocyte pyroptosis. In conclusion, HOTAIR competitively bound to miR-17-5p, relieving its inhibition of RORA transcription and leading to increased RORA expression and suppressed cardiomyocyte pyroptosis in HF models.
摘要：
心力衰竭(HF)是与显著的发病率和死亡率相关的复杂临床综合征。长非编码RNA（lncRNA）的失调与HF的发病机理有关。本研究旨在研究lncRNAHOX转录反义RNA（HOTAIR）在小鼠HF模型中心肌细胞焦亡中的作用。通过横主动脉收缩手术建立小鼠HF模型，并通过用H2O2处理HL-1细胞建立了体外HF细胞模型。通过pcDNA3.1-HOTAIR转染在TAC小鼠和HL-1细胞中过表达HOTAIR。在TAC小鼠中评估心脏功能,HE染色评价心肌变化。免疫组化法检测NLRP3的表达。使用ELISA定量心肌损伤标志物和焦亡相关的炎性细胞因子。通过Western印迹分析NLRP3、裂解的半胱天冬酶-1和GSDMD-N的蛋白质水平。双荧光素酶测定和RNA免疫沉淀用于确认HOTAIR和miR-17-5p之间的结合相互作用。miR-17-5p和RORA。通过在HL-1细胞中过表达miR-17-5p或沉默RORA进行功能拯救实验。HOTAIR在TAC小鼠和H2O2诱导的心肌细胞中表现出降低的表达。HOTAIR过表达改善心功能不全,减轻心肌病理损伤，增强心肌细胞活力，减少心肌损伤和焦亡。HOTAIR与miR-17-5p相互作用以抑制RORA转录。miR-17-5p的过表达或RORA的沉默消除了HOTAIR过表达对心肌细胞焦亡的抑制作用。总之,HOTAIR与miR-17-5p竞争结合，减轻其对RORA转录的抑制作用，并导致HF模型中RORA表达增加和心肌细胞焦亡抑制。
