青蒿素抑制Nod样受体家族包含pyrin结构域蛋白3复合物对心肌缺血再灌注损伤的保护作用

Protective effect of artemisinin inhibiting NOD-like receptor family,p yrin domain containing 3 complex on myocardial ischemia-reperfusion injury

目的探讨青蒿素抑制Nod样受体家族包含pyrin结构域蛋白3(NLRP3)复合物对心肌缺血再灌注损伤的保护作用。方法选用SD大鼠,结扎左冠状动脉前降支,构建大鼠心肌缺血再灌注损伤模型,并随机分为对照组、模型组和青蒿素组,每组10只。其中10只不结扎的大鼠作为假手术组。青蒿素组大鼠建模前灌胃25 mg/kg青蒿素,模型组和对照组建模前灌胃等体积生理盐水。3组大鼠造模1 d后采用生化酶法检测心肌损伤标志物酸激酶(CK-MB)、乳酸脱氢酶(LDH)和肌钙蛋白Ⅰ(cTnⅠ);采用TTC染色分析3组大鼠梗死百分比;采用酶联免疫吸附试验(ELISA)分析白细胞介素(IL)-1β和IL-18的表达水平;采用蛋白质印迹法(Western blot)分析大鼠心肌组织半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-1/NLRP3表达水平。组间比较采用单因素方差分析。结果青蒿素组大鼠血清CK-MB、LDH和cTnⅠ水平[(2424.30±199.06)U/L、(1128.30±118.46)U/L、(35.26±3.56)pg/ml]明显低于模型组[(4105.10±191.55)U/L、(1682.00±155.21)U/L、(73.46±8.67)pg/ml],差异有统计学意义(t=19.240、8.968、12.890,P<0.05)。青蒿素组大鼠心肌梗死百分比[(28.05±4.51)%]明显低于模型组[(46.07±4.27)%],差异有统计学意义(t=9.163,P<0.05)。青蒿素组大鼠血清IL-1β和IL-18水平[(92.80±7.94)、(81.40±4.50)pg/ml]明显低于模型组[(172.60±11.62)、(117.70±7.89)pg/ml],差异有统计学意义(t=17.930、12.640,P<0.05)。青蒿素组大鼠心肌组织Caspase-1和NLRP3表达水平(1.37±0.10、1.34±0.05)明显低于模型组(1.80±0.11、2.00±0.13),差异有统计学意义(t=9.542、14.670,P<0.05)。结论青蒿素通过抑制NLRP3复合物活性,降低IL-1β和IL-18表达水平,进而对大鼠缺血再灌注损伤心肌组织起保护作用。

Objective To investigate the protective effect of artemisinin on myocardial ischemia-reperfusion injury by inhibiting NOD-like receptor family,pyrin domain containing 3(NLRP3)complex.Methods SD rats were randomly divided into control group,model group and artemisinin group,10 rats in each group.Among them,10 rats without ligation were used as the sham operation group.Rats in the artemisinin group were gavaged with 25 mg/kg artemisinin before modeling,and those in the model group and the control group were gavaged with equal volume of normal saline before modeling.At 1st day after the model was established in the three groups,creatine kinase isoenzymes(CK-MB),lactate dehydrogenase(LDH)and cardiac troponinⅠ(cTnⅠ)were detected by biochemical enzyme method.The percentage of infarct in the three groups was analyzed by TTC staining.The expression level of interleukin(IL)-1βand IL-18 was analyzed by enzyme linked immunosorbent assay(ELISA).The expression level of cysteinyl aspartate-specific protease(Caspase)-1 and NLRP3 in rat myocardium was analyzed by Western blotting.Results The levels of serum CK-MB,LDH and CTNⅠin artemisinin group[(2424.30±199.06)U/L,(1128.30±118.46)U/L,(35.26±3.56)pg/ml]were significantly lower than those in model group[(4105.10±191.55)U/L,(1682.00±155.21)U/L,(73.46±8.67)pg/ml,t=19.240,8.968,12.890,P<0.05].The percentage of myocardial infarction in artemisinin group[(28.05±4.51)%]was significantly lower than that in model group[(46.07±4.27)%,t=9.163,P<0.05].Serum IL-1βand IL-18 levels in artemisinin group[(92.80±7.94),(81.40±4.50)pg/ml]were significantly lower than those in the model group[(172.60±11.62),(117.70±7.89)pg/ml,t=17.930,12.640,P<0.05].The expression levels of caspase-1 and NLRP3 in in the artemisinin group(1.37±0.10,1.34±0.05)were significantly lower than those in the model group(1.80±0.11,2.00±0.13,t=9.542,14.670,P<0.05).Conclusion Artemisinin can reduce IL-1 by inhibiting the activity of NLRP3 complexβand the expression level of IL-18 in the myocardial tissue of rats with ischemia-reperfusion injury.