CD38抑制剂通过Sirt3/FoxO1途径抑制心肌缺血/再灌注损伤的研究

闫莉; 寿锡凌; 梁磊; 杨光

doi:10.12125/j.chj.202303067

摘要:

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。

Abstract:

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 T and ROS levels in group B were obvious higher than those in group A (P<0.01), the troponin T and ROS levels in group C were significant lower than those in group B (P<0.01) and the troponin T and ROS levels in group D were higher than those in group C (P<0.01 and 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<0.01 and P<0.05). 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.

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