天麻素抗心肌氧化应激损伤的作用机制研究

Protective effect of gastrodin against myocardial oxidative stress damage

摘要:

 摘要：目的  探讨天麻素在大鼠心肌细胞氧化应激损伤时是否通过线粒体机制发挥心脏保护作用。方法  首先使用过氧化氢H2O2 650 μmol/L处理大鼠心脏组织来源的H9c2心肌细胞，复制氧化应激损伤模型。分别使用50.0、10.0、1.0和0.1 μmol/L天麻素预处理，利用共聚焦显微镜成像技术，检测线粒体膜电位的变化，四甲基偶氮唑盐比色法检测天麻素对细胞存活率的影响，Western blot检测天麻素对糖原合成酶激酶-3β（GSK-3β）、蛋白激酶-B（Akt）活性的影响。结果  不同浓度的天麻素预处理均能减弱H2O2引起的四甲基罗丹明乙酯荧光强度降低程度，且与模型组（0.30±0.25）比较，10.0 μmol/L天麻素预处理组（0.79±0.08）作用最为明显。天麻素（10.0μmol/L）预处理能提高H9c2细胞的存活率，使p-GSK-3β（Ser9）、p-Akt（Ser473）蛋白水平升高，而磷脂酰肌醇-3激酶（PI3K）抑制剂渥曼青霉素可以阻断其发挥作用。结论  天麻素能够减轻由H2O2引发的H9c2心肌细胞氧化应激损伤，其可能是通过PI3K/Akt途径使GSK-3β失活，抑制mPTP开放以发挥心脏保护作用。

Abstract:

Abstract: Objective To investigate whether gastrodin plays a protective role in rat myocardial cell oxidative stress by mitochondrial pathway. Methods H2O2 (650 μmol/L) was used to induce oxidative stress injury of H9c2 cells, and the pretreatment concentration of gastrodin was 50.0, 10.0, 1.0 and 0.1 μmol/L respectively. Laser scanning confocal microscopy (LSCM) was used to detect mitochondrial membrane potential. MTT method was used to test the effect of gastrodin on cell survival rate. Western blot was used to observe the influences of gastrodin on the activity of glycogen synthase kinase 3β (GSK-3β) and protein kinase B (Akt). Results Pretreatment of gastrodin with different concentrations could prevent the reduction of fluorescence intensity of tetramethylrhodamine ethyl ester caused by H2O2, and the 10.0 μmol/L gastrodin pretreatment group had the most obvious effect compared with the model group [(0.79 ± 0.08) vs (0.30 ± 0.25)]. Pretreatment with gastrodin(10.0 μmol/L) improved the survival rate of H9c2 cells (P < 0.05) and increased protein expressions of p-GSK-3β (Ser9) and p-Akt (Ser473); whereas PI3K inhibitor, Wortmannin penicillin (Wort), could block this effect. Conclusions Gastrodin can alleviate oxidative stress damage of H9c2 myocardial cells caused by H2O2. It may inactivate GSK-3β through PI3K/Akt pathway, thereby play a cardiac-protective role through inhibition of mPTP opening.