Danshensu (Dan shen suan A) is an active ingredient of Salvia miltiorrhiza with wide cardiovascular benefit.

Danshensu通过清除自由基减少线粒体膜上的脂质过氧化，并通过降低硫醇氧化抑制线粒体膜的渗透性和传导[1]。Danshensu显著提高H9c2心肌细胞的存活率，并降低乳酸脱氢酶(LDH)的释放。Danshensu在H9c2细胞中增加Akt和细胞外信号相关激酶1/2(ERK1/2)的磷酸化，其保护作用部分被磷脂酰肌醇3'-激酶(PI3K)特异性抑制剂wortmannin或ERK特异性抑制剂U0126所抑制。Danshensu能显著对抗心肌缺血/再灌注(MI/R)损伤，其潜在机制可能通过激活PI3K/Akt和ERK1/2信号通路抑制心肌细胞凋亡。通过激活Akt和ERK信号通路，Danshensu增加Bcl-2表达，并减少Bax、活性caspase-3表达。Danshensu已被证明在改善微循环、抑制活性氧物种的形成、抑制血小板黏附和聚集、保护心肌免受缺血损害、保护内皮细胞免受炎症诱导伤害等方面具有生物活性[2]。

在ISO处理的大鼠中，预先使用Danshensu显著降低了ST段（P<0.001）与仅使用ISO的大鼠相比。其预处理同样显著减少了血清cTnI水平（P<0.001）。因此，Danshensu对ISO诱导的大鼠心肌梗死具有显著的心脏保护作用[1]。在MI/R损伤的大鼠模型中，Danshensu显著减小了心肌梗死面积以及血清中肌酸激酶-MB（CK-MB）、心脏肌钙蛋白（cTnI）的产生[2]。

Measurement of inhibitory activities of MG-132 against 20S proteasome: The reaction mixture for the 20S proteasome inhibitory assay contains 0.1 M Tris-acetate, pH 7.0, 20S proteasome, MG-132, and 25 μM substrate dissolved in dimethyl sulfoxide in a final volume of 1 mL. After incuba tion at 37 °C for 15 minutes, the reaction is stopped by the addition of 0.1 mL of 10% SDS and 0.9 mL of 0.1M Tris acetate, pH 9.0. The fluorescence of the reaction products is measured. To determine the IC50 against 20S proteasome, various concentrations of MG-132 are included in the assay mixture.

Cardiomyocytes are exposed to ischemia by replacing medium with an 'ischemic buffer', this buffer is designed to simulate the extracellular milieu of myocardial ischemia, with the approximate concentrations of potassium, hydrogen, and lactate ions occurring in vivo. Cells are incubated in the hypoxic/ischemic chamber at 37℃ for 2 h in a humidified atmosphere of 5% CO2 and 95% nitrogen. At the onset of reperfusion, cardiomyocytes are randomly exposed to one of the following treatments: vehicle, Danshensu (1 μM or 10 μM), Danshensu plus the PI3K inhibitor wortmannin (10 nM), Danshensu plus the ERK inhibitor U0126 (10 μM). Simultaneously, in the control group, H9c2 cardiomyocytes are cultured under normal conditions in CO2 incubation. (Only for Reference)