【摘要】 目的：探讨特异性长链非编码RNA SLC25a6(long noncoding RNA SLC25a6, lncSLC25a6)在同型半胱氨酸（homocysteine, Hcy）诱导的心肌细胞铜死亡中作用。方法：体外培养大鼠心肌细胞并分为对照（control）与Hcy组，干预细胞48 h后，采用Western blot及免疫荧光染色方法检测铜死亡相关蛋白铁氧还蛋白1(ferredoxin 1,FDX1)、热休克蛋白70(heat shock protein 70, HSP70)等的表达水平；利用荧光染色检测心肌细胞氧化应激状态；采用铜离子试剂盒测定心肌细胞内Cu2+水平；进一步将lncSLC25a6过表达后，分析Hcy对心肌细胞铜死亡相关蛋白表达的影响。结果：与control组相比，80μmol/L Hcy显著加速了心肌细胞损伤，且lncSLC25a6呈低表达（P<0.05）。Western blot检测结果显示：与control组相比，Hcy干预组FDX1表达水平明显降低（P<0.05），而HSP70表达水平明显增高（P<0.05），且Hcy组心肌细胞铜离子表达水平增高（P<0.05）。免疫荧光染色发现，Hcy组FDX1荧光强度明显减弱，HSP70的荧光强度明显增强；进一步过表达lncSLC25a6后，Hcy诱导的心肌细胞铜死亡显著缓解，细胞铜死亡相关蛋白FDX1呈高表达，HSP70呈低表达（P<0.05）。Pearson相关性分析发现，lncSLC25a6的表达水平与FDX1蛋白表达呈负相关（r=-0.676,P=0.046），而与HSP70表达则呈正相关（r=0.680,P=0.044）。结论：lncSLC25a6可显著缓解Hcy诱导的心肌细胞铜死亡，可作为Hcy诱导心肌损伤的潜在治疗靶点。更多还原

【Abstract】 AIM: To investigate the role of specific long noncoding RNA SLC25a6(lncSLC25a6) in homocysteine(Hcy)-induced cuproptosis in cardiomyocytes. METHODS: Rat cardiomyocytes were cultured in vitro and divided into control group and Hcy group. After 48 h of intervention, the expression levels of cuproptosis-related proteins, ferredoxin 1(FDX1) and heat shock protein 70(HSP70), were detected by Western blot and immunofluorescence staining.The oxidative stress state of cardiomyocytes was assessed using fluorescence staining, and the intracellular Cu2+ levels were measured using a copper ion assay kit. Furthermore, the impact of Hcy on the expression of cuproptosis-related proteins in cardiomyocytes was analyzed following overexpression of lncSLC25a6. RESULTS: Compared with the control group, 80μmol/L Hcy significantly accelerated cardiomyocyte damage, with a notable underexpression of lncSLC25a6(P<0. 05).Western blot results indicated that, compared with the control group, the expression level of FDX1 in the Hcy intervention group was significantly reduced(P<0. 05), while the expression level of HSP70 was significantly elevated(P< 0. 05), and the expression level of copper ions in cardiomyocytes of the Hcy group was increased(P<0. 05). Immunofluorescence staining showed a significant reduction in FDX1 fluorescence intensity and a significant increase in HSP70 fluorescence intensity in the Hcy group. Further overexpression of lncSLC25a6 significantly mitigated Hcy-induced cuproptosis in cardiomyocytes, resulting in elevated expression of FDX1 and reduced expression of HSP70(P<0. 05). Pearson correlation analysis demonstrated that the expression level of lncSLC25a6 was negatively correlated with FDX1 protein expression(r=-0. 676, P=0. 046) and positively correlated with HSP70 expression(r=0. 680, P=0. 044). CONCLUSION: lncSLC25a6 significantly mitigates Hcy-induced cuproptosis in cardiomyocytes, positioning it as a potential therapeutic target for managing Hcy-induced cardiac injury.更多还原