迷迭香酸对心肌细胞缺氧复氧损伤的保护作用研究

目的：研究迷迭香酸对大鼠乳鼠原代心肌细胞缺氧复氧损伤的保护作用。 方法：体外培养大鼠原代心肌细胞，复制心肌细胞缺氧复氧损伤模型，分别观察细胞活力和LDH漏出的改变，采用化学发光法检测细胞ATP含量变化，利用荧光探针技术检测细胞内ROS产生的变化，采用流式细胞术及Western blot法进一步考察迷迭香酸对心肌细胞凋亡，cleaved-caspase 3，Akt和p-Akt蛋白表达的影响。 结果：实验结果显示，25，50，100 mg·L^-1迷迭香酸均能显著抑制缺氧复氧导致的细胞活力下降，LDH漏出及ROS的过度产生，并能维持细胞内ATP水平；50，100 mg·L^-1迷迭香酸显著抑制缺氧复氧诱导的心肌细胞凋亡和下调cleaved-caspase 3 蛋白表达，并且100 mg·L^-1迷迭香酸能够明显上调p-Akt 蛋白的表达。 结论：迷迭香酸具有显著的抗心肌细胞缺氧复氧损伤作用，能够改善心肌细胞能量代谢和减少细胞凋亡，其保护作用可能是通过激活Akt通路实现的。

Protective effect of rosmarinic acid on hypoxia/reoxygenation injury in cardiomyocytes

Objective: To study the protective effect of rosmarinic acid (Ros A) on the primary cardiomyocyte hypoxia/reoxygenation (H/R) injury. Method: Primary cardiomyocytes of rats were cultured in vitro to establish the H/R injury of cardiomyocytes and observe the changes in the cell viability and LDH leakage. The changes in ATP content and ROS in cardiomyocytes were measured by using chemiluminescence and fluorescent probe technique. The effects of rosmarinic acid on the apoptosis of cardiomyocytes, cleaved-caspase 3, Akt and p-Akt protein expression were further detected by flow cytometry and western blot analysis. Result: According to the experimental results, Ros A at doses of 25, 50, 100 mg·L^-1 could inhibit the decrease in H/R-induced cell viability, LDH leakage and excessive ROS generation, and maintain the ATP level in cells. Ros A at doses of 50, 100 mg·L^-1 could remarkably inhibit the H/R-induced cardiomyocyte apoptosis and down-regulate the expression of cleaved caspase-3. Moreover, Ros A at doses of 100 mg·L^-1 could significantly up-regulate the expression of p-Akt. Conclusion: Ros A has the significant effect in resisting the cardiomyocyte H/R injury, improve cardiomyocyte energy metabolism and reduce cell apoptosis, which is related to the activation of Akt pathway.