丹红注射液预处理和后处理对家兔心肌缺血再灌注损伤的保护作用

Protective effects of pretreatment and posttreatment with Danhong Injection on myocardial ischemia reperfusion injury of rabbits

目的探讨丹红注射液对家兔心肌缺血再灌注损伤的保护作用,并比较预处理和后处理的效果。方法将32只健康成年雄性家兔随机分为假手术组、缺血再灌注组、预处理组、后处理组,每组8只。预处理组在结扎冠状动脉前10 min、后处理组在再灌注前10 min经耳缘静脉给予丹红注射液2 mL/kg,均缺血30 min后再灌注2 h。记录缺血前、结扎30 min、再灌注1 h、再灌注2 h的左室舒张末压(LVEDP)、左心室收缩压(LVSP)、左室内压最大上升速率(+dp/dtmax)。测定再灌注结束时肌酸激酶(CK)、乳酸脱氢酶(LDH)、肌钙蛋白I(cTnI)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,检测心肌组织中Bcl-2、Bax表达情况,TUNEL法检测心肌细胞凋亡指数。结果结扎30 min、再灌注1 h、再灌注2 h,缺血再灌注组LVSP、+dp/dtmax均明显低于假手术组(P均<0.05),LVEDP均明显高于假手术组(P均<0.05);预处理组和后处理组LVSP和+dp/dtmax均明显高于缺血再灌注组(P均<0.05),LVEDP均明显低于缺血再灌注组(P均<0.05),且预处理组LVSP和+dp/dtmax均明显高于后处理组而LVEDP明显低于后处理组(P均<0.05)。再灌注结束时,缺血再灌注组血清CK、LDH、cTnI、MDA和心肌组织中Bcl-2、Bax蛋白表达水平均明显高于假手术组(P均<0.05),血清SOD明显低于假手术组(P均<0.05);预处理组和后处理组血清CK、LDH、cTnI、MDA和心肌组织中Bax蛋白表达水平均明显低于缺血再灌注组(P均<0.05),血清SOD和心肌组织中Bcl-2蛋白表达水平明显高于缺血再灌注组(P均<0.05),且预处理组血清CK、LDH、cTnI、MDA和心肌组织中Bax蛋白表达水平均明显低于后处理组而SOD、Bcl-2蛋白表达水平均明显高于后处理组(P均<0.05)。心肌细胞凋亡指数缺血再灌注组明显高于假手术组(P<0.05),预处理组和后处理组均明显低于缺血再灌注组(P均<0.05),且预处理组明显低于后处理组(P<0.05)。结论丹红注射液对家兔心肌缺血再灌注损伤有保护作用,且预处理效果优于后处理,其保护作用可能是通过抗氧化和抑制心肌细胞凋亡实现的。

Objective It is to investigate the effect of Danhong Injection on myocardial ischemia-reperfusion injury in rabbits and to compare the effects between the prevention and posttreatment with this medicine. Methods 32 healthy adult male rabbits were randomly divided into 4 groups: sham group,myocardial ischemia reperfusion group, pretreatment group and posttreatment group, 8 rabbits in each group. The rabbits in the pretreatment group and posttreatment group were respectively treated with Danhong Injection (2 mL/kg) 10 minutes before coronary artery ligation or 10 minutes before reperfusion intravenously from ear.All the animals were suffered from myocardial ischemia for 30 minutes and reperfusion for 2 h. Left ventricular end-diastolic pressure (LVEDP), left ventricular systolic pressure (LVSP), and left atrial pressure maximal rate of increase (+dp/dtmax) were recorded before ischemia and after 30 min of ligation, 1 h of reperfusion, and 2 h of reperfusion. The activity of creatine kinase (CK), lactate dehydrogenase (LDH), troponin I (cTnI), superoxide dismutase (SOD) and the content of malondialdehyde (MDA) at the end of reperfusion were determined, the expression of Bcl-2 and Bax in myocardial tissue was detected, and the cardiomyocyte apoptosis index was detected by TUNEL method. Results After ligation for 30 min, reperfusion for 1 h, and reperfusion for 2 h, LVSP and +dp/dtmax were significantly lower and LVEDP were significantly higher in the ischemia-reperfusion group than that in the sham-operated group ( P <0.05). The LVSP and +dp/dtmax of the pretreatment group and the posttreatment group were significantly higher and the LVEDP was significantly lower than those of the ischemia-reperfusion group ( P <0.05), the LVSP and +dp/dtmax were significantly higher and LVEDP was significantly lower in the pretreatment group than those in the posttreatment group ( P <0.05). At the end of reperfusion, the levels of serum CK, LDH, cTnI, MDA and Bcl-2, Bax protein in myocardial tissue in the ischemia-reperfusion group were significantly higher and that of serum SOD was significantly lower than those in the sham operation group ( P <0.05);the levels of serum CK, LDH, cTnI, MDA and Bax expression in myocardial tissue in the pretreatment group and the posttreatment group were significantly lower, and the levels of serum SOD and expression of Bcl-2 protein in myocardial tissue was significantly higher than that in ischemia-reperfusion group ( P <0.05), and the levels of serum CK, LDH, cTnI, MDA and Bax protein in myocardial tissue were significantly lower, the expression levels of SOD and Bcl-2 protein in the pretreatment group were significantly higher than those in the posttreatment group ( P <0.05). The myocardial apoptosis index was significantly higher in the ischemia-reperfusion group than that in the sham operation group ( P <0.05), the indexe in the pretreatment group and posttreatment group were significantly lower than the ischemia-reperfusion group ( P <0.05), and the pretreatment group was lower than the posttreatment group ( P <0.05). Conclusion Danhong injection has protective effect on myocardial ischemia-reperfusion injury in rabbits, and the pretreatment effect is better than post-treatment. The protective effect may be achieved by anti-oxidation and inhibition of cardiomyocyte apoptosis.