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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H_2O_2650μmol/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0.1μmol/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H_2O_2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,10.0μmol/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(Ser~9)、p-Akt(Ser~(473))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H_2O_2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。
Objective To investigate whether gastrodin can play a cardioprotective role in mitochondrial oxidative stress injury in rat cardiomyocytes. Methods H9c2 cardiomyocytes derived from rat heart tissue were treated with H 2 O 2 at 2650 μmol / L, and then oxidative stress injury model was duplicated. 50.0,10.0,1.0 and 0.1μmol / L gastrodin were used for pretreatment. The changes of mitochondrial membrane potential were detected by confocal microscopy. The effect of gastrodin on cell viability was detected by MTT colorimetry Western blot was used to detect the effect of gastrodin on the activity of glycogen synthase kinase-3β (GSK-3β) and protein kinase-B (Akt). Results Gastrodin pretreatment at different concentrations attenuated the decrease of fluorescence intensity of tetramethylrhodamine ethyl ester induced by H_2O_2. Compared with model group (0.30 ± 0.25), pretreatment with 10.0 μmol / L gastrodin pretreatment group (0.79 ± 0.08 The most obvious effect. Gastrodin pretreatment increased the survival rate of H9c2 cells and increased the protein levels of p-GSK-3β (Ser ~ 9) and p-Akt (Ser ~ (473) -3 kinase (PI3K) inhibitor wortmannin can block its function. Conclusion Gastrodin can reduce oxidative stress injury induced by H_2O_2 in H9c2 cardiomyocytes, which may be due to inactivation of GSK-3β via PI3K / Akt pathway and inhibition of mPTP opening to exert cardioprotection.