神经生长因子β预先给药对大鼠离体心脏再灌注时内质网应激性细胞凋亡的影响  

Effects of nerve growth factor-beta pretreatment on endoplasmic reticulum stress-induced cell apoptosis during ischemia-reperfusion in isolated rat hearts

作　　者：魏珂[1] 刘力[1] 谢飞[1] 罗洁[1] 闵苏[1] 

机构地区：[1]重庆医科大学附属第一医院麻醉科,400016

出　　处：《中华麻醉学杂志》2014年第5期541-544,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年基金（81000066）;卫生部国家临床重点专科建设项目[财社（2011）170号];重庆市医学重点学科建设项目[渝卫科教[2007]2号]

摘　　要：目的 评价神经生长因子β（NGF-β）预先给药对大鼠离体心脏缺血再灌注时内质网应激性细胞凋亡的影响.方法 雄性SD大鼠32只,体重180 ～ 220 g,制备Langendorff离体心脏灌注模型.取模型制备成功的心脏32个,采用随机数字表法,将其分为4组（n=8）：正常对照组（C组）、缺血再灌注组（I/R组）、NGF-β预先给药组（N组）和NGF-β联合trkA受体拮抗剂K252a组（N＋K组）.C组用K-H液持续灌注195 min.I/R组先用K-H液平衡灌注45 min,N组和N＋K组用K-H液平衡灌注15 min,再分别用含有0.1 μg/ml NGF-β和0.1 μg/ml NGF-β混合100 nmol/L K252a的K-H液继续灌注30min.除C组外,其余各组均停止灌注30 min和再灌注120 min.于平衡灌注末（基础水平）、再灌注5、30、60和120 min时记录心率（HR）、左室舒张末压（LVEDP）、左室发展压（LVDP）、左室内压最大上升速率（＋dp/dtmax）;再灌注120 min时取心肌组织,测定心肌细胞凋亡指数、葡萄糖调节蛋白78 （GRP78）、C/EBP同源蛋白（CHOP）和caspase-12的表达水平.结果 与C组比较,I/R组和N组HR、LVDP和＋dp/dtmax降低,LVEDP升高,心肌组织细胞凋亡指数、GRP78和CHOP表达升高,I/R组心肌组织caspase-12表达上调（P＜0.05）;与I/R组比较,N组HR、LVDP和＋dp/dtmax升高,LVEDP降低,心肌组织细胞凋亡指数、GRP78、CHOP和caspase-12表达降低,N＋K组心肌组织GRP78表达下调（P＜0.05）,心功能各指标差异无统计学意义（P＞0.05）;与N组比较,N＋K组HR、LVDP和＋dp/dtmax降低,LVEDP升高,心肌组织细胞凋亡指数、GRP78和CHOP表达升高,心肌组织caspase-12表达上调（P＜0.05）.结论 NGF-β预先给药可减轻大鼠离体心脏缺血再灌注损伤,其机制与激活trkA受体后抑制内质网应激性细胞凋亡有关.Objective To evaluate the effects of nerve growth factor-beta （NGF-β） pretreatment on cell apoptosis induced by endoplasmic reticulum stress during ischemia-reperfusion in isolated rat hearts.Methods Male Sprague-Dawley rats,weighing 180-220 g,were anesthetized with intraperitoneal 10% chloral hydrate 300 mg/kg.The hearts were excised and perfused in a Langendorff apparatus with K-H solution aerated with 95% O2 and 5% CO2 at 37 ℃.Thirty-two isolated rat hearts were randomly divided into 4 groups （n =8 each） using a random number table：control group （group C）,I/R group,NGF-β pretreatment group （group N） and NGF-β combined with K252a （trkA receptor antagonist） pretreatment group （group N ＋ K）.In group C,the hearts were continuously perfused with K-H solution for 195 min.The hearts were perfused with K-H solution for 45 min in group I/R.In N and N ＋ K groups,the hearts were perfused with K-H solution for 15 min,and then with K-H solution containing 0.1 μg/ml NGF-β and 0.1 μg/ml NGF-β mixed with 100 nmol/L K252a,respectively,for 30 min.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in I/R,N and N ＋ K groups.HR,left ventricular end-diastolic pressure （LVEDP）,left ventricular developed pressure （LVDP） and ＋ dp/dtmax were measured at the end of 15 min equilibration （baseline） and at 5,30,60 and 120 min of reperfusion.Myocardial specimens were obtained at 120 min of reperfusion for detection of myocardial apoptosis （by TUNEL） and expression of glucose-related protein 78 （GRP78）,CCAAT/enhancer-binding protein homologous protein （CHOP）,and caspase-12 （by Western blot analysis）.Apoptosis index was calculated.Results Compared with group C,HR,LVDP and ＋ dp/dtmax were significantly decreased,and LVEDP,apoptosis index and expression of GRP78 and CHOP were increased in I/R and N groups,and the expression of caspase-12 was upregulated in I/R group.Compared with group I/R,HR,LVDP,and ＋ dp/dtmax were significantly

关 键 词：神经生长因子 内质网 应激 细胞凋亡 心肌再灌注损伤 

分 类 号：R965[医药卫生—药理学]