Southern Medical University, Guangzhou 510515, China.
J Tradit Chin Med. 2010 Dec;30(4):259-64. doi: 10.1016/s0254-6272(10)60053-2.
To investigate the protective action of tanshinone IIA (TSN) on myocardial apoptosis induced by hydrogen peroxide (H2O2) and its effect on prohibitin (PHB) expression to probe the role of PHB in the oxidation stress of myocardial cells.
Primary cultured neonate rat myocardial cells were cultured with TSN (1 x 10(-4) mol/L) for 24 hours, and then the medium was supplemented with 200 micromol/L hydrogen peroxide for 2 h to initiate myocardial cell oxidative stress injury. PHB in myocardial cells was knocked down by small interfering RNA (siRNA), and the expression level of PHB was determined by western blot analysis. Flow cytometry was used to detect the apoptosis rate, intracellular calcium ion concentration ([Ca2+]i) and mitochondrial membrane potential (MMP).
The PHB expression, [Ca2+]i and the apoptotic rate significantly increased, and the MMP significantly decreased in the oxidative stress group compared with the control. The PHB expression, apoptosis rate and [Ca2+]i decreased, and MMP increased significantly in the TSN group compared with the oxidative stress group. Compared with the siRNA negative control group, the PHB expression level in myocardial cells was down-regulated, and the apoptosis rate and [Ca2+]i increased, and MMP decreased significantly in the siRNA group.
TSN can reduce PHB expression in oxidative stress-injured myocardial cells hence protecting the myocardial cells.
研究丹参酮Ⅱ A（TSN）对过氧化氢（H2O2）诱导的心肌细胞凋亡的保护作用及其对抑制素（PHB）表达的影响，探讨 PHB 在心肌细胞氧化应激中的作用。
培养乳鼠心肌细胞，用 TSN（1 x 10(-4) mol/L）孵育 24 小时，然后用 200 μmol/L H2O2 补充培养基 2 小时，诱导心肌细胞氧化应激损伤。用小干扰 RNA（siRNA）敲低 PHB，用 Western blot 分析检测 PHB 的表达水平。用流式细胞术检测细胞凋亡率、细胞内钙离子浓度（[Ca2+]i）和线粒体膜电位（MMP）。
与对照组相比，氧化应激组 PHB 表达、[Ca2+]i 和凋亡率显著增加，MMP 显著降低。与氧化应激组相比，TSN 组 PHB 表达、凋亡率和[Ca2+]i 降低，MMP 显著升高。与 siRNA 阴性对照组相比，siRNA 组心肌细胞 PHB 表达下调，凋亡率和[Ca2+]i 升高，MMP 显著降低。
TSN 可降低氧化应激损伤心肌细胞中 PHB 的表达，从而保护心肌细胞。
