CD38抑制剂通过Sirt3/FoxO1途径抑制心肌缺血/再灌注损伤的研究

CD38 inhibitor inhibits myocardial ischemia-reperfusion injury through Sirt3/FoxO1 pathway

目的探讨CD38抑制剂减轻心肌缺血/再灌注损伤的作用及机制研究。方法雄性SD大鼠20只给予适应性喂养7 d后,随机分为四组,分别为假手术(A)组、心肌缺血/再灌注(B)组、心肌缺血/再灌注+CD38抑制剂(C)组和心肌缺血/再灌注+CD38抑制剂+Sirt3抑制剂(3-TYP)(D)组。B、C、D组建立心肌缺血/再灌注模型。C组和D组用木犀草素(特异性CD38抑制剂)[100 mg/(kg·d)]连续灌胃2周,D组造模前30 min尾静脉注射3-TYP。记录心电图,检测血清肌钙蛋白T水平、ROS水平,测定心肌病变,检测心肌组织中Sirt3、FoxO1、Bax、Bcl-2蛋白表达水平。结果与A组相比,B组心电图ST段明显抬高;与B组相比,C组ST段回落;与C组相比,D组ST段抬高。与A组相比,B组肌钙蛋白T、ROS水平显著增高(P<0.01);与B组相比,C组肌钙蛋白T、ROS水平则显著降低(P<0.01);与C组相比,D组肌钙蛋白T、ROS水平明显增高(P<0.01和P<0.05)。HE染色显示A组心肌排列整齐,无明显炎症浸润,B组结构紊乱,可见炎症细胞浸润,C组和D组较B组排列明显整齐。TUNEL染色结果显示,与A组相比,B组TUNEL阳性细胞比例显著增加(P<0.01);与B组相比,C组的TUNEL阳性细胞比例明显降低(P<0.01);与C组相比,D组的TUNEL阳性细胞比例则显著增加(P<0.01)。与A组相比,B组Sirt3蛋白表达水平显著降低(P<0.01);与B组相比,C组Sirt3蛋白表达水平显著升高(P<0.01);与C组相比,D组Sirt3蛋白表达水平则明显降低(P<0.05)。与A组相比,B组FoxO1蛋白和Bcl-2蛋白表达水平显著降低(P<0.01);与B组相比,C组FoxO1蛋白和Bcl-2蛋白表达水平则显著升高(P<0.01);与C组相比,D组则显著降低(P<0.01和P<0.05)。与A组相比,B组Bax蛋白表达显著升高(P<0.01);与B组相比,C组则显著降低(P<0.01);与C组相比,D组Bax水平则显著升高(P<0.01)。结论CD38抑制剂可能通过Sirt3/FoxO1途径抑制心肌缺血/再灌注引起的心肌损伤。

AIM To investigate the effects of CD38 inhibitors on myocardial ischemia reperfusion injury.METHODS Twenty male Sprague-Dawley rats were given adaptive feeding for 7 days and they were randomly divided into four groups:A.Sham group,B.Myocardial ischemia/reperfusion(I/R)group,C.Myocardial ischemia/reperfusion+CD38 inhibitor group and D.Myocardial ischemia/reperfusion+CD38 inhibitor+Sirt3 inhibitor(3-TYP)group.Myocardial I/R model was established in groups B,C and D.CD38 inhibior Luteolin(100 mg/kg/d)was administered by gavage for 2 weeks in C and D groups,and 3-TYP was injected via tail vein 30min before modeling in D group.Electrocardiogram(ECG),serum troponin T(cTNT)level,ROS level,myocardial lesion and the expressions of Sirt3,FoxO1,Bax and Bcl-2 in myocardial tissue were detected.RESULTS The ECG ST segment elevation in group B was significantly higher than that in group A,ST segment in group C was lower than that in group B and ST segment in group D was higher than that in group C.The troponin I and ROS levels in group B were obvious higher than those in group A(P<0.01),the troponin I and ROS levels in group C were significant lower than those in group B(P<0.01)and the troponin I and ROS levels in group D were higher than those in group C(P<0.01,P<0.05).HE staining showed that the myocardium in group A was arranged neatly and there was no obvious inflammatory infiltration.The structure of group B was disordered and inflammatory cells infiltration was observed.The structures of group C and group D were more orderly than that of group B.TUNEL staining showed that the proportion of TUNEL positive cells in group B was markedly greater than those in group A and group C(P<0.01),and the proportion in group C was much less than that in group D(P<0.01).The protein expression level of Sirt3 in group B was significant lower than those in groups A,C and D(P<0.01)and the expression level in group C was obviously greater than that in group D(P<0.05).The expressions of FoxO1 protein and Bcl-2 protein in group B were much less than those in group A and group C(both,P<0.01),and the expressions in group C was obviously higher than those in group D(P<0.05,P<0.01).However,the expression of Bax protein in group B was notably higher than those in group A and group C(both,P<0.01),and the expression in group C was prominently lower than that in Group D(P<0.01).CONCLUSION CD38 inhibitor inhibits myocardial I/R injury through Sirt3/FoxO1 pathway.