芎芍胶囊联合缺血后适应对大鼠缺血/再灌注心肌MCP-1及TNF-α的影响

Effects of Xiongshao Capsule Combined with Ischemic Postconditioning on Monocyte Chemoattractant Protein-1 and Tumor Necrosis Factor-α in Rat Myocardium with Ischemic Reperfusion Injury

目的观察芎芍胶囊(Xiongshao Capsule,XSC)联合缺血后适应(ischemic postconditioning,IPoC)对大鼠缺血/再灌注(ischemic reperfusion,I/R)损伤心肌单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量及炎性细胞浸润的影响。方法 Spragu-Dawley(SD)大鼠75只,随机分为假手术组、I/R组、IPoC组、福辛普利钠加IPoC组、XSC加IPoC组,每组15只。除假手术组外,其他各组均结扎大鼠冠状动脉前降支30min,再灌注1h。IPoC组大鼠在再灌注1h前予以3次10s的再灌注/缺血循环,福辛普利钠加IPoC组和XSC加IPoC组分别给予福辛普利钠0.9mg/kg和芎芍胶囊0.135g/kg,连续灌胃14天,末次灌胃2h后实施手术取材。采用比色法测定血清肌酸激酶同工酶(creatine kinase-MB,CK-MB)和肌钙蛋白T(cardiac troponin T,cTnT)的含量,氯化硝基四氮唑兰(nitro blue tetrazolium chloride,NBT)染色测定大鼠左室心肌梗死面积,酶联免疫吸附测定法(enzyme-linked immunosorbant assay,ELISA)测定心肌组织MCP-1和TNF-α含量,HE染色观察心肌组织炎性细胞的浸润。结果与I/R组比较,IPoC组大鼠血清CK-MB和cTNT水平明显降低(P<0.01),心肌梗死面积显著减小(P<0.01),心肌组织MCP-1、TNF-α含量和炎性细胞浸润明显下降(P<0.05,P<0.01);与IPoC组比较,XSC加IPoC进一步降低了I/R大鼠心肌梗死面积和CK-MB含量(P<0.01),大鼠心肌MCP-1、TNF-α含量和炎性细胞浸润数量显著降低(P<0.05,P<0.01)。结论芎芍胶囊可加强缺血后适应对I/R大鼠心肌的保护作用,其机制可能与抑制MCP-1、TNF-α表达和炎性细胞浸润有关。

Objective To investigate effect of Xiongshao Capsule （XSC） combined with ischemic postconditioning （IPoC） on tumor necrosis factor-α （TNF-α） and monocyte chemoattractant protein-1 （MCP-1） contents as well as inflammatory cell infiltration （ICI） in myocardium of rat with ischemic reperfusion （I/R） injury.Methods Seventy-five Sprague-Dawley rats were equally randomized into 5 groups,the sham-operated group （A）,the I/R group （B）,the IPoC group （C）,the fosinopril sodium plus IPoC group （D）,and the XSC plus IPoC group （E）.Excepting rats in Group A,all animals received I/R injury through a 30-min occlusion of left anterior descending artery followed by 1-h reperfusion.Additionally,IPoC （3 cycles of 10s reperfusion/10s of ischemia） was applied on rats in Group C before 1h of reperfusion;while rats in Groups D and E were pretreated for 14 days with 0.9mg/kg fosinopril sodium and 0.135g/kg XSC respectively via gastrogavage,and the I/R injury with IPoC applied 2h after the final gavage.Serum creatine kinase-MB （CK-MB） and cardiac troponin T （cTnT） levels were detected by colorimetric method,myocardial infarction size was measured by nitro blue tetrazolium chloride （NBT） staining,MCP-1 and TNF-α contents in myocardial tissue were examined by enzyme-linked immunosorbent assay （ELISA）,and ICI was detected by HE staining.Results Compared with Group B,myocardial enzymes and infarction size were significantly decreased （P0.01）,contents of MCP-1,TNF-α and ICI in myocardial tissue were significantly decreased （P0.05,P0.01） in Group C.Compared with Group C,further reduced infarction size and release of myocardial enzyme CK-MB （P0.01） were seen in Group E,and contents of MCP-1 and TNF-α as well as ICI in myocardial tissue in Group E were also significantly lower （P0.05,P0.01）.Conclusion XSC could enhance the protective effect of IPoC on rat with myocardial I/R injury,and the mechanism may be related to its inhibition on MCP-1 and TNF-α expressions as well as ICI suppression.