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目的 本研究旨在探讨受体相互作用蛋白(RIP)3-钙离子/钙调蛋白依赖蛋白激酶(CaMK)II通路激活的程序性坏死(necroptosis)在糖尿病心肌缺血/再灌注(I/R)损伤中的作用。方法 成年(3~4月龄)雄性C57BL/6小鼠70只，随机分为正常组（n=30）与STZ诱导的糖尿病组（n=40）。随后采用小鼠急性心肌I/R在体模型(缺血30min再灌注4 h)。再灌注后取心肌组织，一部分应用Western blot法检测RIP3-CaMKII信号的表达及修饰变化，另一部分做冰冻切片组织免疫荧光，通过伊文氏蓝阳性区域的面积占比反映程序性坏死的程度。结果 与正常组相比，糖尿病组小鼠I/R后心肌程序性坏死程度显著升高（P<0.05）；在糖尿病I/R心肌中RIP3表达水平和磷酸化CaMKII水平较正常组I/R心肌均显著升高（P<0.05）。对糖尿病小鼠再灌注前给予CaMKII抑制剂KN-93处理可以显著降低糖尿病心肌I/R后的程序性坏死比例（P<0.05）。结论 本研究证实在糖尿病心肌I/R损伤中程序性坏死显著增加，提示RIP3-CaMKII信号增强引发心肌程序性坏死在糖尿病心肌缺血易损中起重要作用。
Objectives This study was aimed to investigate the role of necroptosis activated by the receptor-interacting protein (RIP) 3-Ca 2+ / CaMK II pathway in diabetic myocardial ischemia / reperfusion (I / R The role of injury. Methods Seventy male C57BL / 6 Mice (3 to 4 months old) were randomly divided into normal group (n = 30) and STZ-induced diabetic group (n = 40). Acute myocardial I / R in vivo was then used in vivo (4 h reperfusion after 30 min ischemia). Myocardial tissues were harvested after reperfusion. The expression and modification of RIP3-CaMKII signal were detected by Western blot, and the frozen sections were immunofluorescence. The proportion of positive area of Irwin blue reflected the degree of programmed necrosis. Results Compared with the normal group, the degree of myocardial necrosis in diabetic rats was significantly increased after I / R (P <0.05). The expression of RIP3 and phosphorylated CaMKII in diabetic I / R myocardium were significantly higher than those in normal group Myocardium were significantly increased (P & lt; 0.05). Administration of KN-93, a CaMKII inhibitor, prior to reperfusion in diabetic mice significantly reduced the rate of programmed necrosis following diabetic I / R (P & lt; 0.05). Conclusions This study demonstrates a significant increase in programmed necrosis in diabetic myocardial I / R injury, suggesting that increased RIP3-CaMKII signaling may contribute to myocardial necrosis in diabetic myocardium.