姜黄素对缺血再灌注H9c2心肌细胞凋亡和GSK-3表达及其磷酸化的影响

Effects of Curcumin on Ischemia/Reperfusion Induced Apoptosis of H9c2 Myocardial Cells and the Expression of Glycogen Synthase Kinase-3 and Its Phosphorylation

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中文关键词: 姜黄素心肌细胞缺血再灌注糖原合成酶激酶-3 酪氨酸磷酸化蛋白丝氨酸磷酸化蛋白

英文关键词:curcumin myocyte ischemia/reperfusion glycogen synthase kinase 3 tyrosine-phosphorylated protein serine-phosphorylated protein

基金项目:南京医科大学科技发展基金（No.2011NJMU119）

作者	单位	E-mail
虞燕萍,周承亮,傅云峰
黄先玫	杭州市第一人民医院儿科（杭州310006）	hxianmei630715@163.com

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中文摘要:

目的观察姜黄素对缺血再灌注（ischemia-reperfusion，I/R）H9c2心肌细胞凋亡和糖原合成酶激酶3（GSK-3）表达及其磷酸化的影响。方法利用缺血台氏液对体外培养的H9c2心肌细胞模拟I/R处理，同时随机分为模型组（模拟缺血90 min，再灌注30 min）、姜黄素组（再灌注同时加入7.5 μmol/L姜黄素）和对照组（正常台氏液培养120 min代替模拟I/R），采用流式细胞术检测细胞凋亡情况，采用Western blot检测GSK-3、酪氨酸磷酸化GSK-3 （pTyr-GSK-3）和丝氨酸磷酸化GSK-3 （pSer-GSK-3）蛋白的表达。结果与对照组比较，模型组H9c2细胞凋亡率明显提高（t=10.439， P=0.000），模型组pTyr-GSK-3和pSer-GSK-3表达均显著增加（t=5.208， P=0.006； t=5.854， P=0.004）。与模型组比较，姜黄素组凋亡率及pTyr-GSK-3表达水平显著降低（t=－8.325， P=0.001；t=－3.607， P=0.023），存活率及pSer-GSK-3表达水平显著升高（t=9.165， P=0.001；t=3.747， P=0.02）。结论 GSK-3的酪氨酸和丝氨酸磷酸化可能参与心肌细胞I/R损伤，姜黄素减少I/R心肌细胞凋亡可能与其通过减少酪氨酸磷酸化、增加丝氨酸磷酸化抑制GSK-3活性有关。

英文摘要:

Objective To investigate the effects of curcumin on the apoptosis of ischemia/reperfusion （I/R） induced H9c2 myocardial cells and the expression of glycogen synthase kinase-3 （GSK-3） and its phosphorylation state. Methods I/R of H9c2 cells in vitro was simulated by an ischemic Tyrode solution. Cells were randomly divided into 3 groups， i.e.， the model group （exposed to ischemic solution for 90 min followed by 30 min reperfusion with the normal Tyrode solution）， the curcumin group （7.5 μmol/L curcumin added at the onset of reperfusion for 30 min）， and the control group （exposed to normal Tyrode solution for 120 min）. Then， the cell apoptosis was detected in 3 groups by flow cytometry. The expression levels of GSK-3， phosphotyrosine-GSK-3 （pTyr-GSK-3）， and phosphoserine-GSK-3 （pSer-GSK-3） were detected by Western blot. Results Compared with the control group，the apoptosis rate was obviously enhanced in the model group （t=10.439， P=0.000）. And the relative expression levels of both pTyr-GSK-3 and pSer-GSK-3 significantly increased in the model group （t=5.208， P=0.006； t=5.854， P=0.004， respectively）. Compared with the model group， the apoptosis rate and the expression of pTyr-GSK-3 significantly decreased in the curcumin group （t=－8.325， P=0.001； t=－3.607， P=0.023）. Compared with the model group， the rate of viable cells and the expression of pSer-GSK-3 were significantly enhanced in the curcumin group （t=9.165， P=0.001； t=3.747， P=0.02）. Conclusions Both pTyr-GSK-3 and pSer-GSK-3 might participate in the IR injured myocardial cells. Curcumin could reduce apoptosis of I/R injured myocardial cells， which might be correlated with GSK-3 inhibition by decreasing tyrosine phosphorylation and increasing serine phosphorylation.

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