龙胆苦甙后处理对心脏缺血再灌注损伤的防治作用

Preventive and therapeutic effect of post-treatment with gentiopicroside on myocardial ischemia-reperfusion injury

目的观察龙胆苦甙(GPS)后处理对离体心肌缺血/再灌注损伤(I/R)的防治作用及其可能的机制。方法将30只C57BL6小鼠分为三组,即假手术组(只开胸不结扎冠状动脉左前降支),I/R组以及I/R+GPS后处理组(I/R+GPS组)。采用小鼠在体模型,小鼠麻醉后,开胸短暂结扎冠状动脉左前降支模拟缺血,缺血30 min后松开线结进行再灌注,再灌注前10 min腹腔注射给药。分别于再灌注2 h(Western blot法检测Caspase-3、Bax、Bcl-2)以及24 h(心脏超声测定EF值、TTC伊文氏蓝双染色检测心肌梗死面积以及经颈动脉取血行LDH活性检测)后处死动物。结果与假手术组比较,I/R组EF值降低,心肌梗死面积增加,血清中LDH含量增高,Caspase-3、Bax表达增高,Bcl-2表达降低(P均<0.01)。与I/R组相比,I/R+GPS组的EF值升高,心肌梗死面积减少,血清中LDH含量降低,Caspase-3、Bax表达降低,Bcl-2表达增高(P均<0.01)。结论 GPS后处理可以明显降低缺血再灌注对心脏的损伤,增加心脏收缩能力,增强抗凋亡分子Bcl-2的表达,抑制Bax以及Caspase-3所介导的心肌细胞凋亡。

Objective To investigate the preventive and therapeutic effect of post-treatment with gentiopicroside （GPS） on myocardial ischemia-reperfusion （I/R） injury and the underlying mechanism. Methods We divided 30 C57BI.6 mice into three groups： sham operation group （thoracotomy without ligation of the left anterior descending coronary artery）, I/R group and I/R ＋ GPS post-treatment group. After anesthetization, the left anterior descending coronary arter- ies of C57 mice were temporally ligatured by a slipknot for 30 minutes to mimic ischemia situation. Ten minutes before reperfusion, the GPS was given. After 2 h and 24 h, the mice were sacrificed. In addition, at 2 h, Western blotting was used to detect the expression of caspase-3, Bax and Bcl-2, and at 24 h, ejection fraction （EF） was determined by cardiac ultrasound, myocardial infarction size was detected by TI＇C Evans blue staining and LDH activity detection was conducted in the carotid artery blood. Results Compared with the sham group, the EF was declined, the infarction size was en- larged, the serum LDH amount was increased, the expression of caspase-3 and Bax was up-regulated, while Bcl-2 expres- sion was down-regulated in the I/R group （all P 〈 0.01 ）. Compared with the I/R group, the ejection fraction recovered, the infarction size and the LDH level were declined, the expression of apoptotic relative protein, such as caspase-3 and Bax was down-regulated, and the expression of Bcl-2 was un-regulated in the I/R ＋ GPS group （ all P 〈0.01 ）. Conclusion Post-treatment with GPS can significantly reduce the ischemia-reperfusion damage to the heart, increase cardiac contractili- ty, enhance the expression of Bcl-2, and inhibit Bax and caspase-3 mediated apoptosis of myocardial cells.