Department of Geriatrics, Taihe Hospital, Hubei University of Medicine, No. 32 Renminnan Road, Shiyan City, 442000, Hubei Province, China.
Department of Cardiology, Taihe Hospital, Hubei University of Medicine, No. 32 Renminnan Road, Shiyan City, 442000, Hubei Province, China.
J Thromb Thrombolysis. 2021 Oct;52(3):720-729. doi: 10.1007/s11239-021-02506-3. Epub 2021 Jun 28.
Cardiovascular diseases rank the top cause of morbidity and mortality worldwide and are usually associated with blood reperfusion after myocardial ischemia/reperfusion injury (MIRI), which often causes severe pathological damages and cardiomyocyte apoptosis. LSINCT5 expression in the plasma of MI patients (n = 53), healthy controls (n = 42) and hypoxia-reoxygenation (HR)-treated cardiomyocyte AC16 cells was examined using qRT-PCR. The effects of LSINCT5 on cell viability and apoptosis were detected by MTT and flow cytometry, respectively. The expression of apoptosis-related proteins Bcl2, Bax and caspase 3 were tested by Western blot. The interaction between LSINCT5 and miR-222 was predicted by bioinformatic analysis. Moreover, changes in viability and apoptosis of AC16 cells co-transfected with siLSINCT5 and miR-222 inhibitor after HR treatment were examined. At last, the expression of proteins in PI3K/AKT pathway, namely PTEN, PI3K and AKT, was examined to analyze the possible pathway participating in LSINCT5-mediated MI/RI. Our study showed that LSINCT5 expression was upregulated in the plasma of MI patients and HR-treated AC16 cells. LSINCT5 overexpression significantly decreased cell viability and apoptosis. Luciferase reporter gene assay and RNA pulldown assay showed that LSINCT5 was a molecular sponge of miR-222. MiR-222 silencing in AC16 cells simulated the phenotypes of MIRI patients and HR-treated cells, indicating that LSINCT5 functions via miR-222 to regulate proliferation and apoptosis of HR-treated AC16 cells. We also showed that proteins of PI3K/AKT signaling pathway were affected in HR-treated AC16 cells, and LSINTC5 knockdown rescued these effects. LncRNA LSINCT5 was upregulated during MI pathogenesis, and LSINCT5 regulated MIRI possibly via a potential LSINCT5/miR-222 axis and PI3K/AKT signaling pathway. Our findings may provide novel evidence for MIRI prevention.
心血管疾病是全球发病率和死亡率最高的疾病,通常与心肌缺血/再灌注损伤(MIRI)后的血液再灌注有关,这通常会导致严重的病理损伤和心肌细胞凋亡。使用 qRT-PCR 检测 MI 患者(n=53)、健康对照者(n=42)和缺氧-复氧(HR)处理的心肌细胞 AC16 中血浆中的 LSINCT5 表达。通过 MTT 和流式细胞术分别检测 LSINCT5 对细胞活力和细胞凋亡的影响。通过 Western blot 检测凋亡相关蛋白 Bcl2、Bax 和 caspase 3 的表达。通过生物信息学分析预测 LSINCT5 与 miR-222 的相互作用。此外,还检测了 HR 处理后共转染 siLSINCT5 和 miR-222 抑制剂的 AC16 细胞活力和细胞凋亡的变化。最后,检测 PI3K/AKT 通路中蛋白质的表达,即 PTEN、PI3K 和 AKT,以分析参与 LSINCT5 介导的 MI/RI 的可能通路。我们的研究表明,LSINCT5 在 MI 患者和 HR 处理的 AC16 细胞的血浆中表达上调。LSINCT5 过表达显著降低细胞活力和细胞凋亡。荧光素酶报告基因测定和 RNA 下拉测定表明,LSINCT5 是 miR-222 的分子海绵。AC16 细胞中 miR-222 的沉默模拟了 MIRI 患者和 HR 处理细胞的表型,表明 LSINCT5 通过 miR-222 调节 HR 处理的 AC16 细胞的增殖和凋亡。我们还表明,HR 处理的 AC16 细胞中 PI3K/AKT 信号通路的蛋白受到影响,而 LSINTC5 敲低可挽救这些作用。在 MI 发病机制中,lncRNA LSINCT5 上调,LSINCT5 通过潜在的 LSINCT5/miR-222 轴和 PI3K/AKT 信号通路调节 MIRI。我们的研究结果可能为 MIRI 的预防提供新的证据。