丹参酮ⅡA对H9c2心肌细胞缺血再灌注损伤的保护机制

Mechaniam of the protective effect of Tanshinone ⅡA on myocardial ischemia-reperfusion injury

目的观察丹参酮ⅡA对心肌缺血再灌注损伤的保护作用及相关信号传导通路。方法通过建立H9c2心肌细胞缺血再灌注模型,分别加入不同浓度丹参酮ⅡA,采用CCK-8检测细胞存活率,通过流式细胞术检测细胞凋亡率;另外分为丹参酮ⅡA组、AG490组、丹参酮ⅡA及AG490组,通过Western blot方法检测JAK2、P-JAK2、STAT3、P-STAT3蛋白表达。结果 CCK-8检测显示模型组细胞存活率为78.90%±5.163%,与对照组比较,差异有统计学意义(P<0.05);丹参酮ⅡA 2.5μM组细胞存活率为85.76%±6.101%,与模型组比较,P>0.05;丹参酮ⅡA 10μM组细胞存活率为90.62%±2.321%,与模型组比较,P<0.05;丹参酮ⅡA 40μM组细胞存活率为86.38%±4.712%,与模型组比,P<0.05;流式细胞术检测显示加入丹参酮ⅡA缺血再灌注导致的心肌细胞凋亡数减少;丹参酮ⅡA组P-JAK2、P-STAT3蛋白表达较缺血再灌注组明显上升,而AG490组的P-JAK2蛋白表达明显下调。结论丹参酮ⅡA可改善缺血再灌注引起的大鼠心肌细胞凋亡,其保护机制可能与JAK2/STAT3信号通路有关。

Objective To study the protective effect of tanshinone ⅡA on myocardial ischemia-reperfution（I/R） injury and related signialing pathway. Methods Made H9c2 myocytes I/R models, add different concentrations of tanshinone ⅡA after 24 hours, then test apoptosis rate by cck-8 and flow cytometry. Add tanshinone ⅡA, AG490, tanshinone ⅡA＋AG490 respectively, then assay the expression of JAK2, P-JAK2, STAT3, P-STAT3 by western blot. Results CCK-8tests showed the cell survival rate of model group was 78.90%±5.163%, compared with the control group had statistical differences. Compared with the model group, the cell survival rate increased to 85.76% ±6.101% at the tanshinone ⅡA concentration of 2.5 μM（P0.05）, increased to 90.62%±2.321% at the tanshinone ⅡA concentration of 10 μM（P0.05）, and increased to 86.38%±4.712% at the tanshinone ⅡA concentration of 40 μM（P0.05）. Flow cytometry analysis also displayed tanshinone ⅡA could decrease the number of myocardial apoptosis which caused by I/R injury. The expression of P-JAK2 and P-STAT3 increased in tanshinone ⅡA group compared to I/R group. The expression of PJAK2 reduced in AG490 group. Conclusion Tanshinone ⅡA can reduce I/R induced myocardial apoptosis, and the protective mechanism may be related to the JAK2-STAT3 signaling pathway.