法舒地尔对大鼠心肌缺血再灌注氧化损伤的保护作用

Protective Effect of Fasudil on Myocardial Ischemia-reperfusion Injury in Rats

目的探究法舒地尔对于大鼠心肌缺血再灌注氧化损伤的影响。方法选择SD大鼠48只,随机分成6组,每组8只。对照组,对大鼠不进行手术;假手术组,对大鼠进行手术但不进行缺血处理;I/R组,大鼠进行缺血1h,再灌注3h处理;法舒地尔低、中、高剂量三种处理组,即大鼠在手术前1h分别用10、25和50mg·kg-1法舒地尔水溶液进行处理。采用结扎左冠状动脉前降支的方法制备在体大鼠心肌缺血再灌注损伤模型。分别检测6组大鼠心肌梗死面积、肌酸激酶活性、心脏组织心肌细胞凋亡Caspase-3活性。Real-time PCR用于检测心肌组织中NADPH氧化酶2(NOX2)mRNA的表达。Western blot实验检测心肌肌球蛋白轻链2v(MLC2v)磷酸化以及NOX2的蛋白表达。检测H2O2产生以及Rho激酶(ROCK),肌球蛋白轻链磷酸酶(MLCP)和NOX活性。结果I/R组心肌纤维化和紊乱加重,血清中肌酸激酶(CK)活性增加,I/R处理之后大鼠心脏TUNEL阳性细胞增加,ROCK活性升高,而MLCP活性降低(P均<0.05);胞核中I/R处理p-MLC2v水平增加,NOX2的mRNA以及蛋白水平在I/R处理之后在大鼠心脏中上调,并且NOX活性以及H2O2产物增加(P均<0.05),而法舒地尔处理组能够抑制上述现象,呈剂量依赖性(P<0.05)。结论法舒地尔能够减轻缺血再灌注对大鼠心肌的损伤,与抑制氧化损伤有关。

Objective To explore the protective effect of fasudil on myocardial ischemia-reperfusion injury in rats.Methods Forty-eight SD rats were selected and randomly divided into 6 groups with 8 rats in each group.SD rats were randomly divided into six groups:control group,without any surgery on rat;the sham group,rats underwent surgical procedures without ischemic results;the I/R group,rats were subjected to 1h-ischemia followed by 3h-reperfusion;the fasudil-treating group,the fasudil at low(L),medium(M)or high(H)dose group,rats were treated with 10,25 or 50mg·kg-1 fasudil H2O solution 1h before surgery,respectively.The rat model of myocardial ischemia-reperfusion injury was prepared by ligation of the left anterior descending coronary artery.The infarct area of the cardiac tissue,creatine kinase activity,Caspase-3 activity,Hydrogen peroxide,ROCK,MLCP and NOX activities were measured in the six different groups.TUNEL assay was used to evaluate cardiomyocyte apoptosis.Real-time PCR was used to determine NOX2 mRNA expression in cardiac tissues.Western blot was used to detect cytoplasmic and nuclear phosphorylation of MLC2v and expression of NOX2.Results Myocardial fibrosis and disorder were aggravated in I/R group,creatine kinase(CK)activity in serum was significantly increased,TUNEL positive cells in rat heart were significantly increased after I/R treatment,ROCK activity in rat heart was significantly increased after I/R treatment(P<0.05),while MLCP activity was significantly decreased,p-MLC2v level in nucleus was increased after I/R treatment,NOX2 gene expression and protein water were increased.After I/R treatment,the level of NOX and the production of H2O2 in rat heart increased significantly(P<0.05).Fasudil treatment could significantly inhibit the above phenomena in a dose-dependent manner(P<0.05).Conclusion Fasudil can alleviate myocardial damage induced by ischemia-reperfusion in rats which is associated with inhibition of oxidative damage.