Yu Yan, Wang Shuo-Ren, Sun Yi-Kun
Key Laboratory of Chinese Internal Medicine of Ministry of Education, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 May;32(5):661-5.
To explore changes of mitochondrial structure and functions, as well as the protection of ligustrazine in the process of myocardial hypertrophy.
Neonatal myocardial cells were isolated and cultured with angiotensin II (Ang II) for 72 or 96 h. The total protein content was detected using BCA method. The cell diameter was measured by inverted microscope, by which to reflect the proliferation situation of cardiomyocytes. The mitochondrial membrane potential (MMP) was measured by fluorescence microscope. The mitochondrial monoamine oxidase (MAO) activity was detected by spectrophotometer. The mitochondrial cytochrome oxidase (COX) activity and the mitochondrial damage percentage were detected by microplate reader, by which to reflect the damage of mitochondrial outer membrane's structure and the membranes' function. Also, cells were treated with ligustrazine and losartan and then the pharmacological effects on the mitochondrial structure and functions in the myocardial cells treated with Ang II were observed.
At 72 h and 96 h, when compared with the blank group, cells treated with Ang II had increased total protein content (P < 0.01) and enlarged diameter (P < 0.01). Treated with Ang II, the MAO activity and the outer membrane damage percentage of myocardial cells significantly increased (P < 0.01), and mitochondrial COX activity and the mitochondrial MMP significantly decreased (P < 0.01). Compared with the model group at the same time period, ligustrazine significantly reduced myocardial cells' total protein content and myocardial cell diameter, and significantly decreased myocardial cells' MAO activity, increased mitochondrial COX activity, improved the outer membrane damage percentage and inner membrane MMP at 72 and 96 h, all showing statistical difference (P < 0.01, P < 0.05).
During the process of myocardial hypertrophy existed the damage to the mitochondrial structure and functions. Ligustrazine protected the mitochondrial structure and functions of the myocardial cells in reversing Ang II induced myocardial cell hypertrophy.
探讨心肌肥厚过程中线粒体结构和功能的变化以及川芎嗪的保护作用。
分离新生大鼠心肌细胞，用血管紧张素II（Ang II）培养72或96小时。采用BCA法检测总蛋白含量。用倒置显微镜测量细胞直径，以反映心肌细胞的增殖情况。用荧光显微镜测量线粒体膜电位（MMP）。用分光光度计检测线粒体单胺氧化酶（MAO）活性。用酶标仪检测线粒体细胞色素氧化酶（COX）活性和线粒体损伤百分比，以反映线粒体外膜结构和膜功能的损伤情况。同时，用川芎嗪和氯沙坦处理细胞，观察其对Ang II处理的心肌细胞线粒体结构和功能的药理作用。
在72小时和96小时时，与空白组相比，Ang II处理的细胞总蛋白含量增加（P<0.01），直径增大（P<0.01）。经Ang II处理后，心肌细胞的MAO活性和外膜损伤百分比显著增加（P<0.01），线粒体COX活性和线粒体MMP显著降低（P<0.01）。与同期模型组相比，川芎嗪在72小时和96小时时显著降低心肌细胞总蛋白含量和心肌细胞直径，显著降低心肌细胞MAO活性，增加线粒体COX活性，改善外膜损伤百分比和内膜MMP，均有统计学差异（P<0.01，P<0.05）。
在心肌肥厚过程中存在线粒体结构和功能的损伤。川芎嗪在逆转Ang II诱导的心肌细胞肥厚过程中保护心肌细胞的线粒体结构和功能。
