RIP3-CaMKⅡ通路激活程序性坏死在糖尿病心肌缺血/再灌注损伤中的作用  

RIP3-CaMKⅡ pathway is involved in activating necroptosis in diabetic myocardial ischemia/reperfusion injury

作　　者：薛涵 李晨[2] 杨铮[2] 石曌玲[1] 刘曼玲[2] 殷玥[1,2] 马恒[1,2] 

机构地区：[1]第四军医大学基础部生理学教研室,陕西西安710032 [2]第四军医大学基础部病理生理学教研室,陕西西安710032

出　　处：《心脏杂志》2018年第2期125-129,共5页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(81322004,31671424);陕西省社发攻关项目资助(2011KJXX66,2015KW-050)

摘　　要：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30 min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。AIM To investigate the role of necroptosis activated by RIP3-CaMKII pathway under conditions of diabetic myocardial ischemia/reperfusion. METHODS Male C57BL/6 mice （aged 3 -4 months） were randomized into diabetes and normal control groups. For the diabetes group, a model of type 1 diabetes mellitus was employed. Mice were subjected to a myocardial I/R model utilizing anterior descending artery ligation for 30 minutes followed by d-hour reperfusion. Myocardial tissue was excised after reperfusion. Western blot was used to detect RIP3 and phospho-CaMKII expression and cryosection and immunofluorescence were used to evaluate necroptosis by Evans-blue dye positive area percentage. RESULTS Compared with the normal I/R group, the diabetic I/R group showed enhanced necroptosis indicated by a higher EBD-positive area percentage （P 〈 0. 05 ） and increased expression of RIP3 and phospho-CaMKII （P 〈 0.05）. KN-93, an inhibitor diabetic I/R group （P 〈 0.05 ）. CONCLUSION of CaMKII, significantly reduced necroptosis in the Necroptosis is enhanced in diabetic myocardial I/R injury, implying that enhanced the RIP3-CaMKII pathway, which activates myocardial necroptosis, plays an important role in diabetic myocardial I/R.

关 键 词：糖尿病 心肌缺血/再灌注 程序性坏死 RIP3 CAMKII 

分 类 号：R331.3[医药卫生—人体生理学]