AMPK通路在硫化氢后处理减轻2型糖尿病大鼠心肌缺血-再灌注损伤中的作用

发布时间：2018-12-09 18:48

【摘要】：目的观察硫化氢后处理对2型糖尿病大鼠心肌缺血-再灌注(ischemia reperfusion,IR)损伤的效果,探讨AMP依赖的蛋白激酶(AMPK)通路在其中发挥的作用。方法雄性SD大鼠84只,1月龄,体重100~150g,采用随机数字表法分为六组:假手术组(Sham组)、IR组、AMPK抑制剂compound c组(CC组)、compound c溶剂组(DMSO组)、硫化氢后处理组(NaHS组)和CC+NaHS组,每组14只。成功建立糖尿病模型后,Sham组仅开胸但不结扎阻断血流;IR组在开胸分离左冠状动脉前降支后用止血钳夹紧圈套管缺血30min,松开圈套管再灌注4h后取心脏;CC组于术前1h腹腔注射CC 250μg/kg,其余操作同IR组;DMSO组于术前1h给予同样剂量的DMSO,其余操作同IR组;NaHS组于开放左冠状动脉1min内快速静脉注射NaHS 0.05mg/kg,再灌注4h;CC+NaHS组:于术前1h腹腔注射CC,随后于冠状动脉左前降支阻断30min,同样于开放左冠状动脉1min内快速静脉注射NaHS 0.05mg/kg,再灌注4h。之后对所有动物进行安乐死,采用TTC染色法测定心肌梗死范围,Western blot法测定AMPK、LC3、p62蛋白含量。结果与Sham组比较,其余五组再灌注4h时大鼠心肌梗死范围明显增加,AMPK、LC3、p62蛋白含量明显升高(P0.05);与IR组比较,NaHS组大鼠再灌注4h时心肌梗死范围明显减小(P0.05),AMPK蛋白含量明显升高,LC3、p62蛋白含量明显降低(P0.05);与NaHS组比较,CC+NaHS组再灌注4h时心肌梗死范围明显增加,AMPK水平明显降低,LC3、p62蛋白表达含量明显升高(P0.05)。CC组与DMSO组上述各指标差异无统计学意义。结论硫化氢后处理能减轻2型糖尿病大鼠缺血-再灌注损伤后心肌梗死,其机制与调节自噬小体清除及修复AMPK通路调控的自噬流有关。
[Abstract]:Objective to observe the effects of hydrogen sulfide post-treatment on myocardial ischemia reperfusion (ischemia reperfusion,IR) injury in type 2 diabetic rats and to explore the role of AMP dependent protein kinase (AMP) (AMPK) pathway. Methods Eighty-four male SD rats were randomly divided into six groups (Sham group (), IR group), AMPK inhibitor compound c group (CC group), compound c solvent group (DMSO group), sham operation group (), IR group) and AMPK inhibitor compound c group (DMSO group). Hydrogen sulfide group (NaHS group) and CC NaHS group (14 rats in each group). After the diabetic model was established successfully, the Sham group only opened the chest without ligation to block the blood flow, and in the IR group, the left anterior descending coronary artery was separated with a hemostatic clamp for 30 minutes after ischemia, and the heart was taken after reperfusion for 4 hours after releasing the trap tube. CC 250 渭 g / kg was injected intraperitoneally 1 hour before operation in CC group, and the rest was treated with the same dose of DMSO, at 1 hour before operation in DMSO group, IR group in DMSO group, NaHS 0.05 mg / kg in NaHS group in open left coronary 1min for 4 h after reperfusion. In the CC NaHS group, CC, was injected intraperitoneally 1 hour before operation and then blocked at the left anterior descending coronary artery for 30 minutes, and NaHS 0.05mg / kg was injected intravenously into the open left coronary 1min for 4 h after reperfusion. After that, all animals were euthanized, and the content of AMPK,LC3,p62 protein was determined by TTC staining method, Western blot method. Results compared with Sham group, the myocardial infarction size and the content of AMPK,LC3,p62 protein in the other five groups were significantly increased at 4 h after reperfusion (P0.05). Compared with IR group, the myocardial infarction size of NaHS group decreased significantly at 4 h reperfusion (P0.05). (P0.05) the content of), AMPK protein increased significantly, and the content of LC3,p62 protein decreased significantly (P0.05). Compared with NaHS group, the myocardial infarction size of, CC NaHS group increased significantly, the level of AMPK decreased and the expression of LC3,p62 protein increased significantly at 4 h after reperfusion in, CC NaHS group (P0.05). There was no significant difference between). CC group and DMSO group (P0.05). Conclusion hydrogen sulfide post-treatment can alleviate myocardial infarction after ischemia-reperfusion injury in type 2 diabetic rats, and its mechanism is related to regulation of autophagy clearance and repair of autophagy regulated by AMPK pathway.
【作者单位】：苏州大学附属第二医院麻醉科;
【基金】：江苏省自然科学基金面上研究项目(BK20141187) 苏州市科技计划项目(SYS201473,SS201613) 苏州大学科研预研基金项目(SDY2015A18)
【分类号】：R587.2

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