OGT介导的RIPK3糖基化在七氟醚后处理减轻离体大鼠心肌缺血再灌注损伤中的作用  

Effects of O-N-acetyl-glucosamine transferase-mediated receptor interacting protein kinase 3 O-GlcNAcylation on ischemia reperfusion injury in rats after sevoflurane postconditioning

作　　者：张静[1] 王城 余鹏[3] 唐晓祎 田龙 邓小兵 Zhang Jing;Wang Cheng;Yu Peng;Tang Xiaoyi;Tian Long;Deng Xiaobing(Department of Anesthesiology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Anesthesiology,Nanchang Hongdu Hospital of Traditional Chinese Medicine,Nanchang 330038,China;Department of Metabolism and Endocrinology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Anesthesiology,Lushan Rehabilitation Center of PLA Joint Logistics Support Force,Nanchang 332000,China)

机构地区：[1]南昌大学第二附属医院麻醉科,南昌330006 [2]南昌市洪都中医院麻醉科,南昌330038 [3]南昌大学第二附属医院内分泌科,南昌330006 [4]解放军联勤保障部队庐山康复疗养中心麻醉科,南昌332000

出　　处：《国际麻醉学与复苏杂志》2022年第6期571-578,共8页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金(82160371);江西省自然科学青年基金(20212BAB216051)。

摘　　要：目的:评估O-N-乙酰葡糖胺(O-N-acetyl-glucosamine, O-GlcNAc)糖基化修饰转移酶(O-GlcNAc transferase, OGT)介导的受体相互作用蛋白(receptor interacting protein kinase, RIPK) 3糖基化在七氟醚后处理减轻离体大鼠心肌缺血再灌注(ischemia reperfusion, IR)损伤中的作用。方法:取Langendorff离体灌注模型制备成功的大鼠心脏45个,采用随机数字表法分为5组(每组9个):假手术组(SHAM组)、IR组、七氟醚后处理组(SPC组)、SPC+溶剂组[SPC+二甲基亚砜(dimethyl sulfoxide, DMSO)组]和SPC+OGT抑制剂组(SPC+OSMI-1组)。建模完成后各组均平稳30 min,之后SHAM组给予K-H液持续灌注150 min,其余各组大鼠心脏均经历停止灌注30 min后恢复灌注2 h的过程;SPC组、SPC+DMSO组和SPC+OSMI-1组均于再灌注初给予含2.4%七氟醚的K-H液持续灌注15 min;此外,在术前准备的K-H液中,SPC+DMSO组给予50 μmol/L DMSO, SPC+OSMI-1组给予50 μmol/L OSMI-1。连续监测各组大鼠缺血前即刻(T 0)、再灌注30 min (T 1)、再灌注60 min (T 2)、再灌注90 min (T 3)、再灌注2 h (T 4)的左室峰压(left ventricular peak pressure, LVSP)、左室舒张末压(left ventricular end-diastolic pressure, LVEDP)、心率、左室压力升高最大速率(maximum rate of rise of left ventricular pressure, +dp/dt max)与左室压力降低最大速率(maximum rate of drop of left ventricular pressure,-dp/dt max);在再灌注末,采用1%氯化三苯基四氮唑 (triphenyl tetrazolium chloride, TTC)染色检测各组大鼠心肌梗死范围;采用ELISA法检测各组大鼠冠状动脉漏出液乳酸脱氢酶(lactate dehydrogenase, LDH)浓度;Western blot法检测各组大鼠心脏O-GlcNAc、OGT、O-糖基化糖苷酶(O-GlcNAcase, OGA)、磷酸化受体相互作用蛋白1 (phosphorylated RIPK1, p-RIPK1)、磷酸化RIPK3 (phosphorylated RIPK3, p-RIPK3)和磷酸化混合系激酶结构域样蛋白(phosphorylated mixed lineage kinase domain-like protein, p-MLKL)蛋白水平。 结果:与T 0比较,IR组、SPC组、SPC+DMSO组Objective To evaluate the effect of O-N-acetyl-glucosamine(O-GlcNAc)transferase(OGT)mediated receptor interacting protein kinase 3(RIPK3)O-GlcNAcylation on ischemia reperfusion injury(IR)in rats after sevoflurane postconditioning.Methods A total of 45 rat hearts were successfully prepared by Langendorff through the in vitro perfusion modeling method.According to the random table method,they were randomly divided into five groups(n=9):a sham operation group(group SHAM),an ischemia reperfusion group(group IR),a sevoflurane postconditioning group(group SPC),a SPC+solvent group[group SPC+dimethyl sulfoxide(DMSO)]and a SPC+OGT inhibitor group(group SPC+OSMI-1).After modeling,each group kept stable for 30 min,then group SHAM was given K-H solution for 150 min,and the rat hearts were stopped perfusion for 30 min followed by 2 h reperfusion in all groups expect group SHAM.In group SPC,group SPC+DMSO and group SPC+OSMI-1,K-H solution containing 2.4%sevoflurane were continuously perfused at the first 15 min of reperfusion.In addition,in the K-H solution prepared before surgery,group SPC+DMSO was given 50µmol/L DMSO,while group SPC+OSMI-1 was given 50µmol/L OSMI-1.Then,the left ventricular peak pressure(LVSP),left ventricular end-diastolic pressure(LVEDP),heart rate,the maximum rate of rise of left ventricular pressure(+dp/dtmax),and the maximum rate of drop of left ventricular pressure(-dp/dtmax)immediately before ischemia(T0),after reperfusion for 30 min(T1),60 min(T2),90 min(T3),and 2 h(T4).At the end of reperfusion,the myocardial infarction area was measured by 1%2,3,5 triphenyltetrazolium chloride(TTC)staining.The concentration of serum lactate dehydrogenase(LDH)was detected by enzyme-linked immunosorbent assay(ELISA).The levels of O-GlcNAc,OGT,O-GlcNAcase(OGA),p-RIPK1,p-RIPK3 and phosphorylated mixed lineage kinase domain-like protein(p-MLKL)were measured by Western blot.Results Compared with those at T0,group IR,group SPC,group SPC+DMSO,and group SPC+OSMI-1 showed remarkable decreases in heart rate,LVSP,+dp/dtmax a

关 键 词：糖基化 心肌 缺血再灌注损伤 麻醉药 吸入 

分 类 号：R614[医药卫生—麻醉学]