本研究主要探讨miR-652-3p在异氟烷(ISO)抗心肌缺血再灌注(I/R)损伤中的作用及机制。H9c2细胞用不同浓度的ISO进行预处理,随后,建立缺氧/复氧(H/R)模型。miR-652-3p的水平,ISLLIMhomeobox1(ISL1),通过逆转录聚合酶链反应(RT-qPCR)评估炎性细胞因子白细胞介素(IL)-6和肿瘤坏死因子-α(TNF-α)。采用酶联免疫吸附试验检测心肌损伤标志物的浓度,如肌酸激酶-MB(CK-MB)和心肌肌钙蛋白I(cTnI)。细胞计数试剂盒-8用于评估细胞活力,而流式细胞术用于测量细胞凋亡。此外,我们进行了双荧光素酶报告基因试验,以验证ISL1和miR-652-3p之间的靶向关系.在这里,我们证实miR-652-3p的水平随着缺氧时间的延长而逐渐增加;然而,ISO预处理抑制了这种增加(P<0.05)。此外,ISO预处理防止了细胞活力的降低,细胞凋亡增加,和IL-6,TNF-α的过量生产,CK-MB,和H/R诱导的cTnI(P<0.05)。然而,ISO的抑制作用被miR-652-3p水平的升高所抵消(P<0.05)。ISL1是miR-652-3p的潜在靶标。与对照相比,H/R诱导抑制了ISL1水平,ISO处理后其表达增加(P<0.05)。ISL1过表达抑制了ISO对miR-652-3p升高所致心肌损伤保护作用的消除(P<0.05)。这项研究的发现证实,miR-652-3p通过靶向ISL1减弱了ISO对心肌缺血心肌细胞的保护作用。
This research is concentrated on investigating the role and mechanism of miR-652-3p in the protective effects of isoflurane (ISO) against myocardial ischemia-reperfusion (I/R) injury. H9c2 cells underwent pretreatment with varying concentrations of ISO, and subsequently, a hypoxia/reoxygenation (H/R) model was constructed. The levels of miR-652-3p, ISL LIM homeobox 1 (ISL1), and inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-α) were evaluated through reverse transcription polymerase chain reaction (RT-qPCR). Enzyme-linked immunosorbent assay was employed to investigate concentrations of myocardial injury markers, such as creatine kinase-MB (CK-MB) and cardiac troponin I (cTnI). Cell counting kit-8 was used to evaluate cell viability, while flow cytometry was utilized to measure apoptosis. Additionally, a dual luciferase reporter assay was conducted to validate the targeting relationship between ISL1 and miR-652-3p. Herein, we confirmed that the level of miR-652-3p was gradually increased with prolonged hypoxia; nevertheless, this increase was suppressed by ISO pretreatment (P < 0.05). Additionally, ISO pretreatment prevented the decrease in cell viability, increase in apoptosis, and overproduction of IL-6, TNF-α, CK-MB, and cTnI induced by H/R (P < 0.05). However, the inhibitory effects of ISO were counteracted by the increased levels of miR-652-3p (P < 0.05). ISL1 is a potential target of miR-652-3p. H/R induction suppressed ISL1 levels compared to the control, but ISO treatment increased its expression (P < 0.05). Overexpression of ISL1 inhibited the elimination of the protective effect of ISO on myocardial damage induced by the elevation of miR-652-3p (P < 0.05). The findings of this research confirm that miR-652-3p attenuated the protective effect of ISO on cardiomyocytes in myocardial ischemia by targeting ISL1.