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目的:探讨川芎嗪、参附注射液及其联合应用预处理对心肌缺血再灌注损伤大鼠左室前壁缺血处的心肌组织热休克蛋白70(HSP70),p38有丝分裂原激活蛋白激酶(p38 MAPK)表达及抗氧化能力的保护作用。方法:Wister大鼠被随机分为5组:假手术组(sham)与缺血再灌注模型组(IR):生理盐水ip连续3 d;川芎嗪注射液预处理组(LI):川芎嗪注射液术前连续3 d ip(20 mg·kg-1·d-1);参附注射液预处理组(SFI):参附注射液术前连续3 d ip(10 mg·kg-1·d-1);川芎嗪联合参附注射液预处理组(LI+SFI):川芎嗪(20 mg·kg-1·d-1)+参附注射液(10 mg·kg-1·d-1)术前连续3 d ip。制备大鼠心肌缺血再灌注损伤模型,心肌组织缺血30 min,再灌注120 min取左室前壁缺血处的心肌组织制备10%匀浆取上清液。应用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,二硫代二硝基甲苯法测定谷胱甘肽过氧化物酶(GSH-Px)活性,应用免疫组化链霉菌抗生物素蛋白-过氧化物酶连结(S-P)法检测左室前壁缺血处的心肌组织热休克蛋白70(HSP70)和p38有丝分裂原激活蛋白激酶(p38 MAPK)蛋白的表达。结果:IR组较sham组SOD,GSH-Px活性显著下降(P<0.01),HSP70,p38 MAPK阳性表达显著增强(P<0.01)。LI组较IR组,SOD活性显著升高(P<0.01)和GSH-Px活性升高(P<0.05),HSP70显著升高(P<0.01),P38MAPK蛋白阳性表达显著减弱(P<0.01)。SFI较IR组,SOD,GSH-Px活性升高(P<0.05),HSP70升高(P<0.05),p38 MAPK表达降低(P<0.05)。LI+SFI组较IR组,SOD,GSH-Px活性显著升高(P<0.01),HSP70显著升高(P<0.01),p38 MAPK蛋白阳性表达显著减弱(P<0.01)。LI+SFI组较LI组(P<0.05)、SFI组(P<0.01)SOD活性增高,LI+SFI组较SFI组GSH-Px活性增高(P<0.05),HSP70阳性表达增强(P<0.05),p38 MAPK阳性表达减弱(P<0.05)。结论:川芎嗪、参附注射液预处理均可拮抗心肌缺血再灌注损伤,尤以川芎嗪联合参附注射液效果更明显。机制可能与增加SOD,GSH-Px活性、增强HSP70表达和抑制p38 MAPK表达有关。
Objective: To investigate the effects of ligustrazine, shenfu injection and their combined pretreatment on the expression of heat shock protein 70 (HSP70), p38 mitogen-activated protein kinase (p38) in myocardial ischemia-reperfusion injury myocardium, p38 MAPK) expression and anti-oxidative capacity of the protective effect. Methods: Wister rats were randomly divided into 5 groups: Sham group and ischemia-reperfusion model group (IR): normal saline ip for 3 days; Ligustrazine injection pretreatment group (LI): Ligustrazine injection The rats in the Shenfu injection pretreatment group (SFI) were given ip (10 mg · kg-1 · d · d-1) for 3 consecutive days before the operation -1); Ligustrazine combined with Shenfu injection pretreatment group (LI + SFI): Ligustrazine (20 mg · kg -1 · d -1) + Shenfu injection (10 mg · kg -1 · d -1 ) For 3 days before surgery ip. Preparation of rat myocardial ischemia-reperfusion injury model, myocardial ischemia 30 min, 120 min reperfusion left ventricular anterior myocardial ischemia in the preparation of 10% homogenate to take the supernatant. The activity of superoxide dismutase (SOD) was measured by xanthine oxidase method. The activity of glutathione peroxidase (GSH-Px) was determined by dithiobisnitrotoluene method. Immunohistochemical streptavidin The expression of heat shock protein 70 (HSP70) and p38 mitogen - activated protein kinase (p38 MAPK) protein in myocardium of left anterior ischemic area were detected by protein - peroxide enzyme linked immunosorbent assay (SP). Results: The activities of SOD and GSH-Px in IR group were significantly lower than those in sham group (P <0.01), and the positive expressions of HSP70 and p38 MAPK were significantly increased (P <0.01). Compared with IR group, the activity of SOD and the activity of GSH-Px in LI group were significantly increased (P <0.01), HSP70 (P <0.01), P38MAPK protein was significantly decreased (P <0.01) . Compared with IR group, SFI increased the activity of SOD and GSH-Px (P <0.05), HSP70 increased (P <0.05) and the expression of p38 MAPK decreased (P <0.05). The activity of SOD and GSH-Px in LI + SFI group was significantly higher than that in IR group (P <0.01), HSP70 was significantly increased (P <0.01), p38 MAPK protein expression was significantly decreased (P <0.01). The activity of GSH-Px in LI + SFI group was higher than that in LI group (P <0.05), and the activity of SOD in SFI group was higher than that in SFI group (P <0.05) ), p38 MAPK positive expression decreased (P <0.05). CONCLUSION: Tetramethylpyrazine and Shenfu injection can both prevent myocardial ischemia-reperfusion injury, especially ligustrazine and Shenfu injection. Mechanism may be related to increasing SOD, GSH-Px activity, enhancing HSP70 expression and inhibiting p38 MAPK expression.