基于自噬探讨芬太尼复合丙泊酚对大鼠心脏缺血再灌注损伤的影响

The effects of fentanyl combined with propofol on heart ischemia-reperfusion injury in rats via autophagy

目的基于自噬探讨芬太尼复合丙泊酚对大鼠心脏缺血再灌注(I/R)损伤的影响。方法健康雄性SPF级SD大鼠30只,随机分为5组:假手术组、模型组、芬太尼治疗组、丙泊酚治疗组和芬太尼+丙泊酚治疗组。除假手术组外,其余大鼠均建立心肌缺血再灌注损伤模型,芬太尼治疗组、丙泊酚治疗组和芬太尼+丙泊酚治疗组在造模前分别用20μg/kg芬太尼、50 mg/kg丙泊酚以及20μg/kg芬太尼复合50 mg/kg丙泊酚预处理。实验结束后30 min处死大鼠,取心脏组织做后续检测。通过HE和TUNEL染色观察大鼠心脏组织病理损伤,TTC染色观察大鼠心肌梗塞面积,电镜观察心肌自噬泡的形成情况,Western blot检测自噬相关蛋白Beclin1、ATG5和LC3B的表达情况。结果与假手术组相比,模型组大鼠心肌病理变化损伤严重,心脏细胞凋亡与梗死面积百分比升高,心肌细胞线粒体发生肿胀及自噬相关蛋白Beclin1、ATG5和LC3B-II表达增加;与模型组相比,20μg/kg芬太尼、50 mg/kg丙泊酚以及20μg/kg芬太尼复合50 mg/kg丙泊酚预处理均显著减少了I/R大鼠心肌病理损伤,降低了心脏细胞凋亡与梗死面积百分比,并减轻了心肌细胞线粒体肿胀程度及降低了自噬相关蛋白Beclin1、ATG5和LC3B-II的表达,20μg/kg芬太尼复合50 mg/kg丙泊酚预处理在其中的表现最好,而芬太尼和丙泊酚之间没有显著差异。结论芬太尼和丙泊酚均能抑制心肌细胞自噬和线粒体肿胀,同时减轻心肌I/R损伤,20μg/kg芬太尼与50 mg/kg丙泊酚对心肌I/R损伤的治疗效果没有显著差异,此外,芬太尼和丙泊酚联合使用对心肌I/R损伤有协同作用。

Objective To investigate the effect of fentanyl combined with propofol on cardiac autophagy and cardiac ischemia reperfusion injury(I/R)in rats.Methods Thirty healthy male SPF SD rats were randomly divided into five groups:the sham group,model group,fentanyl group,propofol group and fentanyl+propofol group.The I/R model was established for all groups except the sham group.The fentanyl group,propofol group and fentanyl+propofol treatment group were pretreated with 20μg/kg fentanyl,50 mg/kg propofol and 20μg/kg fentanyl combined with 50 mg/kg propofol,respectively.After 30 min,the rats were sacrificed and heart tissue was collected for analyses.HE and TUNEL staining were used to observe the pathological injury of rat heart tissue,TTC staining was used to observe the area of myocardial infarction and electron microscopy was used to observe mitochondrial structure.Western blot was used to detect the expression of autophagy-related proteins Beclin1,ATG5 and L3B.Results Myocardial pathological changes were observed in the model group but not the sham group;in addition,the percentage of cardiac cell apoptosis and infarct area were increased,mitochondrial swelling in myocardial cells was observed and the expression of autophagy-related proteins Beclin1,ATG5 and L3B-II were increased.Treatment of 20μg/kg fentanyl,50 mg/kg propofol and 20μg/kg fentanyl combined with 50 mg/kg propofol pretreatment significantly reduced myocardial pathological injury,cardiac apoptosis and percentage of infarct area in I/R rats.The treatments alleviated mitochondrial swelling in myocardial cells and decreased the expression of autophagy-related proteins Beclin1,ATG5 and L3B-II.Pretreatment of 20μg/kg fentanyl combined with 50 mg/kg propofol showed the best performance among the three treatments;there was no significant difference between fentanyl and propofol.Conclusions Both fentanyl and propofol inhibit autophagy,alleviate mitochondrial swelling and reduce myocardial I/R injury.There was not a significantly different effect between 20μg/kg fentanyl and 50 mg/kg propofol in the treatment of myocardial I/R injury.In addition,the combination of fentanyl and propofol had a synergistic effect on myocardial I/R injury.