参附注射液对糖尿病大鼠缺血再灌注损伤心肌DJ-1表达的影响

Effect of Shenfu injection on expression of DJ-1 in ischemia reperfusion myocardium of diabetic rats

目的观察参附注射液(SFI)对糖尿病大鼠缺血再灌注损伤(IR)心肌DJ-1表达的影响,并探究其对糖尿病IR心肌保护作用的分子机制。方法健康雄性SD大鼠腹腔注射60 mg/kg链尿佐菌素制备糖尿病模型。取糖尿病造模成功大鼠30只,随机数字表法分为三组(n=10):假手术组(S组)、IR组、IR+SIF组。心肌IR模型采用结扎冠脉左前降支(LAD)30 min后松开120 min制备,S组只穿线包绕LDA而不结扎。IR+SFI组开胸前SFI以10 m L·kg-1·h-1持续泵注,开胸后以3 m L·kg-1·h-1持续输注至手术结束,其余两组泵注等量晶体液。检测SD大鼠心梗面积、血清肌酸激酶-MB(CK-MB)含量、心肌DJ-1表达,超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与S组比较,IR组大鼠的心梗面积[(49.0±5.3)%vs(0.0±0.0)%],CK-MB[(1 982±275)U/L vs(1 076±262)U/L]、MDA[(13.1±1.9)nmol/mg vs(7.6±1.1)nmol/mg]含量显著增加,差异均有统计学意义(P<0.05),而DJ-1表达[(0.65±0.14)vs(1.0±0.0)]和SOD活性[(79.0±19.3)U/mg vs(143±15.4)U/mg]显著降低,差异均具有统计学意义(P<0.05);与IR组相比,IR+SFI组大鼠的心梗面积[(35.7±5.3)%vs(49.0±5.3)%],CK-MB[(1 364±228)U/L vs(1 982±275)U/L]和MDA[(7.3±1.25)nmol/mg vs(13.1±1.9)nmol/mg]含量显著降低,差异均有统计学意义(P<0.05),而DJ-1表达[(0.9±0.14)vs(0.65±0.14)]和SOD活性[(119.4±14.6)U/mg vs(79.0±19.3)U/mg]显著降低,差异均有统计学意义(P<0.05)。结论 SFI能显著降低糖尿病心肌IR损伤的易损性,其机制可能与其上调心肌DJ-1表达、增强内源性抗氧化应激有关。

Objective To investigate the effect of Shenfu injection(SFI) on expression of DJ-1 in diabetic ischemia reperfusion(IR) myocardium, and explore the mechanisms of its cardioprotection. Methods Type 1 diabetes models were induced by a single intraperitoneal of 60 mg/kg streptozotocin in healthy male SD rats(weighing200-220 g), and confirmed by fasting blood glucose ≥16.7 mmol/L. Thirty diabetic rats were randomly assigned into3 groups(n=10 each): sham group(S group), IR group and IR+SFI group. Myocardium IR model was established by occlusion of the anterior descending branch of left coronary artery(LAD) for 30 min followed by 120 min reperfusion.LAD was exposed but not occluded in S group. In IR + SFI group, SFI was continuously pumped into femoral vein with 10 m L·kg-1·h-1before opening chest, and after that, it was continuously pumped with 3 m L·kg-1·h-1. Crystal injection was pumped in parallel in the other two groups. After 120 min reperfusion, hearts were removed for determination of infarct size, DJ-1 expression, superoxide dismutase(SOD) activity and malonaldehyde(MDA) release. Blood was collected for creatine kinase-MB(CK-MB) assay. Results Compared with S group, larger myocardium infarct size, higher CK-MB and MDA release, lower DJ-1 expression and SOD activity were detected in IR group,(49.0 ±5.3)% vs(0.0±0.0)%,(1 982±275) U/L vs(1 076±262) U/L,(13.1±1.9) nmol/mg vs(7.6±1.1) nmol/mg,(0.65±0.14) vs(1.0±0.0),(79.0±19.3) U/mg vs(143±15.4) U/mg, P<0.05. Compared with IR group, SFI significantly reduced myocardium infarct size, CK-MB and MDA release, and increased DJ-1 expression and SOD activity in IR + SFI group,(35.7±5.3)% vs(49.0±5.3)%,(1 364±228) U/L vs(1 982±275) U/L,(7.3±1.25) nmol/mg vs(13.1±1.9) nmol/mg,(0.9±0.14) vs(0.65±0.14),(119.4±14.6) U/mg vs(79.0±19.3) U/mg, P<0.05. Conclusion SFI can decrease the vulnerability of diabetic myocardium to IR injury. The mechanism is probably involving in DJ-1 activation, and then enhancing the ability of endogenous antioxidation.