Li Xue-Li, Liu Jian-Xun, Li Peng, Zheng Yong-Qiu
Zhongguo Zhong Yao Za Zhi. 2014 May;39(10):1897-901.
To study the protective effect of rosmarinic acid (Ros A) on the primary cardiomyocyte hypoxia/reoxygenation (H/R) injury.
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.
According to the experimental results, Ros A at doses of 25, 50, 100 mg x 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 x 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 x L(-1) could significantly up-regulate the expression of p-Akt.
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.
研究迷迭香酸(Ros A)对原代心肌细胞缺氧/复氧(H/R)损伤的保护作用。
体外培养大鼠原代心肌细胞,建立心肌细胞H/R损伤模型,观察细胞活力及乳酸脱氢酶(LDH)漏出情况的变化。采用化学发光法和荧光探针技术检测心肌细胞中三磷酸腺苷(ATP)含量及活性氧(ROS)的变化。通过流式细胞术和蛋白质免疫印迹分析进一步检测迷迭香酸对心肌细胞凋亡、半胱天冬酶3(cleaved-caspase 3)、蛋白激酶B(Akt)及磷酸化蛋白激酶B(p-Akt)蛋白表达的影响。
实验结果显示,25、50、100 mg·L⁻¹剂量的Ros A可抑制H/R诱导的细胞活力下降、LDH漏出及过量ROS生成,并维持细胞内ATP水平。50、100 mg·L⁻¹剂量的Ros A可显著抑制H/R诱导的心肌细胞凋亡,并下调cleaved caspase-3的表达。此外,100 mg·L⁻¹剂量的Ros A可显著上调p-Akt的表达。
Ros A对抵抗心肌细胞H/R损伤、改善心肌细胞能量代谢及减少细胞凋亡具有显著作用,其作用机制与激活Akt信号通路有关。