Yin Chao, Huang Chen-Xing, Pan Le, Jin Ke-Jia, Wang Ying, Cao Meng-Ying, Lin Hong, Gao Pan, Li Na, Gong Hui, Zou Yun-Zeng
Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China.
Jinlin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, 130117, China.
Curr Med Sci. 2024 Dec;44(6):1091-1096. doi: 10.1007/s11596-024-2956-1. Epub 2024 Dec 4.
Myocardial ischemia-reperfusion (I/R) injury is associated with a significant reduction in the mitochondrial membrane potential (MMP, ΔΨm). Fluorescence-based assays are effective for labelling active mitochondria in living cells; their application in heart tissue, however, represents a challenge because of a low yield of viable cardiomyocytes after cardiac perfusion. This study aimed to examine a novel method for detecting the changes in the MMP of mouse heart tissue following I/R injury.
The I/R model was established, which was characterized by distinct ischemic area and apoptosis in heart tissue. The MMP was detected via a confocal microscope after the ascending aorta was clamped and the mitochondrial probe solution (containing Mito-Tracker Deep Red FM) was perfused from the apex via a peristaltic pump.
This method enabled the distribution of the probe solution throughout the cardiac tissue via the coronary circulation. Fluorescence detection revealed that the MMP was profoundly reduced in both ischemic area and border area following I/R when compared with that in the sham group. There was no obvious difference in the MMP of the remote area between the I/R group and the sham group.
This study presents a novel method for detecting the MMP in heart tissue, and this method will facilitate the evaluation of changes in the MMP in different regions following I/R.
心肌缺血再灌注（I/R）损伤与线粒体膜电位（MMP，ΔΨm）的显著降低有关。基于荧光的检测方法可有效标记活细胞中的活性线粒体；然而，由于心脏灌注后存活心肌细胞的产量较低，其在心脏组织中的应用面临挑战。本研究旨在探讨一种检测小鼠心脏组织I/R损伤后MMP变化的新方法。
建立I/R模型，其特征为心脏组织中有明显的缺血区域和凋亡。在夹闭升主动脉后，通过蠕动泵从心尖灌注线粒体探针溶液（含Mito-Tracker Deep Red FM），然后用共聚焦显微镜检测MMP。
该方法可使探针溶液通过冠状动脉循环分布于整个心脏组织。荧光检测显示，与假手术组相比，I/R后缺血区域和边缘区域的MMP均显著降低。I/R组与假手术组远隔区域的MMP无明显差异。
本研究提出了一种检测心脏组织中MMP的新方法，该方法将有助于评估I/R后不同区域MMP的变化。
