参元丹对大鼠心肌缺血预适应心肌梗死面积及一氧化氮合酶、蛋白激酶C的影响

目的通过检测大鼠缺血心肌梗死面积的大小、血清一氧化氮合酶(NOS)变化、蛋白激酶C(PKC)的表达,探讨参元丹的药理性预适应作用及其机制。方法选取清洁级健康成年Wistar大鼠60只,雌雄各半,体重(274±42)g,鼠龄4月左右,随机分为6组,分别为对照组、再灌注组、假性缺血预处理组、缺血预处理组、药物预处理+缺血预适应组、药物预处理组,每组10只。分别造模、取血、取心肌组织。将心肌组织浸入10%甲醛溶液中固定,脱水,封蜡,切片,染色。应用HE染色方法对组织切片进行染色,4倍镜下采用彩色病理图像分析系统分析心肌梗死面积。将所取血液离心后取血清,用紫外分光光度法检测血清总NOS的活力和分NOS的活力。应用蛋白激酶C(PKC)免疫组化试剂盒对组织切片进行处理,20倍镜下观察PKC表达情况。结果参元丹能减少大鼠的心肌梗死面积,并与缺血预适应有协同作用,能够加强彼此的抗心肌缺血的能力。各组间血清NOS的变化差异无统计学意义(P0.05)。缺血预处理组、药物预处理+缺血预适应组和药物预处理组的PKC有明显表达,其余各组没有表达。结论参元丹能够减少心肌梗死的面积,具有药理性预适应样心肌保护作用,其作用机制可能与PKC介导有关。

Effect of Shenyuandan on Myocardial Infarct Size, Nitric Oxide Synthase and Protein Kinase C of Myocardial Ischemic Preconditioning Rat

Objective By detecting the infarct size of myocardial ischemia in rats, the changes of serum nitric oxide synthase （NOS） and the expression of protein kinase C （PKC）, to discuss the role of Shenyuandan pharmacological preconditioning and its mechanisms. Method Sixty clean healthy adult Wistar rats half male and half female, （274±42）g body weight, 4 months old, were randomly divided into 6 groups, respectively for the control group, reperfusion group, pseudo-ischemic preconditioning group, ischemic preconditioning group, drug pretreatment ＋ ischemic preconditioning group and drug pretreatment group, 10 rats in each group. Then made the modle, drawn blood, and took the cardiac muscle organization separately. The cardiac muscle organization was plunged into 10% formaldehyde solution to be fixed, dehydrated, wax sealed, slice cut and dyed. HE staining method was used to dye the tissue slice and the colored pathological image analysis system was used to analyze the cardiac arrest area. The activity of serum total NOS and sub-NOS was detected by ultraviolet spectrophotometry. PKC immunohistochemical tissue kit was applied to deal with the tissue and PKC expression situation was observed. Result Shenyuandan could reduce myocardial infarct size in rats, and synergistic effect of ischemic prrconditioning, so as to strengthen their ability of anti-myocardial ischemia. There was no significantly difference in serum NOS among groups （P 〉0.05）. There were obvious expression of PKC in ischemic preconditioning group, drug pretreatment ＋ ischemic preconditioning group and drug pretreatment group, but the rest groups not. Conclusion Shenyuandan was able to reduce the infarct size of myocardial ischemia in rats and had a pharmacological preconditioning-like myocardial protection, its mechanism may be related to PKC-mediated mechanism.