摘要
目的探讨心肌肥大过程中有关心肌细胞线粒体结构和功能的改变,以及川芎嗪的保护作用。方法提取和培养乳鼠心肌细胞,并与血管紧张素Ⅱ(AngⅡ)共培养72、96h。通过BCA法检测细胞总蛋白含量,倒置显微镜拍摄并测量细胞直径,反映心肌细胞增殖情况;通过荧光显微镜测量线粒体内膜膜电位,酶标仪检测线粒体单胺氧化酶(monoamine oxidase,MAO)活性,分光光度计检测线粒体细胞色素C氧化酶(cytochromec oxidase,COX)活性和线粒体损伤百分率等反映心肌细胞线粒外膜结构和内外膜功能的损伤。在此基础上给予川芎嗪与对照药缬沙坦,分析其对心肌细胞重构中线粒体结构和功能的药理作用。结果在72、96h时,模型组较空白组心肌细胞总蛋白含量均增加(P<0.01),心肌细胞直径均增大(P<0.01)。在该增殖过程中,心肌细胞MAO活性和线粒体外膜损伤百分率均显著升高(P<0.01),COX线粒体细胞活性和线粒体膜电位均显著减低(P<0.01)。与模型组同期比较,川芎嗪在72、96h时能显著减少心肌细胞总蛋白质含量及心肌细胞直径,并显著降低心肌细胞MAO活性,提高线粒体COX活性,改善线粒体外膜损伤百分率和内膜膜电位,差异均有统计学意义(P<0.01,P<0.05)。结论在心肌肥大过程中,存在线粒体的结构和功能的损害;川芎嗪在逆转AngⅡ所引起的心肌细胞肥大的过程中具有保护心肌细胞线粒体结构和功能的作用。
Objective To explore changes of mitochondrial structure and functions, as well as the protec- tion of ligustrazine in the process of myocardial hypertrophy. Methods Neonatal myocardial cells were isolated and cultured with angiotensin （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 cyto-chrome 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. Results 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 signifi- cantly 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）. Cenelusions 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.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2012年第5期661-665,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
北京中医药大学自主选题在读研究生资助基金资助项目(No.JYB22-XS073)
关键词
川芎嗪
血管紧张素Ⅱ
心肌肥大
线粒体损伤
ligustrazine
angiotensin Ⅱ
myocardial hypertrophy
mitochondrial damage
