Chen Zhian, Liu Tianying, Xiong Lihui, Liu Zhi
School of Integrated Chinese Medicine, Changchun University of Chinese Medicine, Changchun, 130117, China.
School of Basic Medical Sciences, Changchun University of Chinese Medicine, Changchun, 130117, China.
Cardiovasc Toxicol. 2025 Apr 17. doi: 10.1007/s12012-025-09993-3.
Coronary reperfusion therapy is the most common surgical treatment for myocardial infarction, but it can further induce myocardial ischemia-reperfusion injury (MIRI). Therefore, MIRI following coronary intervention is a challenging clinical issue. This study aims to investigate the involvement of HIF- 1α/BNIP3-mediated mitophagy in the protective effects of Shen-fu Injection (SFI) on MIRI in rats. Key targets and signaling pathways of myocardial MIRI were analyzed using high-throughput transcriptome data from the GSE240842 dataset in the GEO database.To establish the MIRI rat model, the left anterior descending coronary artery was ligated for 30 min, followed by reperfusion for 120 min. Hypoxia/reoxygenation (H/R) in neonatal rat primary cardiomyocytes was induced by oxygen-glucose deprivation for 4 h, followed by reoxygenation for 2 h. Two hours after reperfusion, assessments included myocardial infarction area, CK-MB, CTnI, HE staining, TUNEL, mitochondrial ultrastructure and autophagosomes, HIF- 1α, BNIP3, LC3B-II, LC3B-I protein expression, immunofluorescence, and qRT-PCR. Cardiac function was also evaluated using M-mode ultrasound 2 h after reperfusion. In cardiomyocytes, CCK- 8, EdU cell proliferation levels, scratch assay, mitochondrial membrane potential, ROS levels, cardiomyocyte apoptosis, protein expression levels, and immunofluorescence were assessed 2 h after reoxygenation. Our results indicate that HIF- 1α and BNIP3 are key targets in MIRI. SFI upregulates HIF- 1α expression, promoting moderate mitophagy. This process clears excessively damaged mitochondria, reduces cardiomyocyte apoptosis, and decreases myocardial injury. Additionally, SFI reduces autophagosome accumulation, lowers ROS production, and stabilizes membrane potential. Consequently, the area of myocardial infarction is reduced, and cardiac function is improved. SFI activates the HIF- 1α/BNIP3 pathway to mediate moderate mitophagy, effectively reducing cardiomyocyte apoptosis and alleviating myocardial ischemia-reperfusion injury, thereby protecting cardiomyocytes.
冠状动脉再灌注治疗是心肌梗死最常见的外科治疗方法，但它可进一步诱发心肌缺血-再灌注损伤（MIRI）。因此，冠状动脉介入治疗后的MIRI是一个具有挑战性的临床问题。本研究旨在探讨HIF-1α/BNIP3介导的线粒体自噬在参附注射液（SFI）对大鼠MIRI保护作用中的参与情况。利用GEO数据库中GSE240842数据集的高通量转录组数据分析心肌MIRI的关键靶点和信号通路。为建立MIRI大鼠模型，结扎左冠状动脉前降支30分钟，随后再灌注120分钟。通过氧糖剥夺4小时诱导新生大鼠原代心肌细胞缺氧/复氧（H/R），随后再灌注2小时。再灌注2小时后，评估指标包括心肌梗死面积、肌酸激酶同工酶（CK-MB）、肌钙蛋白I（CTnI）、苏木精-伊红（HE）染色、末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）、线粒体超微结构和自噬体、HIF-1α、BNIP3、微管相关蛋白1轻链3-II（LC3B-II）、微管相关蛋白1轻链3-I（LC3B-I）蛋白表达、免疫荧光和定量逆转录聚合酶链反应（qRT-PCR）。再灌注2小时后还使用M型超声评估心脏功能。在心肌细胞中，复氧2小时后评估细胞增殖-毒性检测试剂盒（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）细胞增殖水平、划痕试验、线粒体膜电位、活性氧（ROS）水平、心肌细胞凋亡、蛋白表达水平和免疫荧光。我们的结果表明，HIF-1α和BNIP3是MIRI中的关键靶点。SFI上调HIF-1α表达，促进适度的线粒体自噬。这一过程清除过度受损的线粒体，减少心肌细胞凋亡，并减轻心肌损伤。此外，SFI减少自噬体积累，降低ROS产生，并稳定膜电位。因此，心肌梗死面积减小，心脏功能得到改善。SFI激活HIF-1α/BNIP3通路以介导适度的线粒体自噬，有效减少心肌细胞凋亡并减轻心肌缺血-再灌注损伤，从而保护心肌细胞。
