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损伤的影响.方法 雄性SD大鼠,8~ 10周龄,体重200~300 g,制备Langendorff离体心脏灌注模型.取模型制备成功的心脏24个,采用随机数字表法,将其分为3组(n=8):缺血再灌注组(I/R组)、NGF-β预先给药组(NGFPt组)、NGF-β预处理组(NGFPc组).各组先用K-H液平衡灌注10 min;I/R组K-H液继续灌注30 min,NGFPt组用含0.1 μg/ml NGF-β的K-H液继续灌注20 min,NGFPc组用含0.1 μg/ml NGF-β的K-H液继续灌注20 min,再用K-H液冲洗10 min;各组均停灌30 min,K-H液复灌120 min.于平衡灌注末、停灌前即刻、再灌注5、30、60和120 min(T1-6)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);于T1和T3-6时收集冠脉流出液2 ml,测定肌酸激酶(CK-MB)、乳酸脱氢酶(LDH)的活性;T6时取心肌组织测定心肌细胞凋亡情况,观察心肌病理学改变.结果 与T1时比较,各组T3-6时+ dp/dtmax降低,LVEDP、冠脉流出液CK-MB、LDH的活性升高,I/R组T3-6时LVDP、HR降低,NGFPt组T3,4时和NGFPc组T3-6时LVDP降低,NGFPt组和NGFPc组T2-6时HR升高(P<0.05).与I/R组比较,NGFPt组和NGFPc组LVDP、+dp/dtmax和HR升高,LVEDP、冠脉流出液CK-MB、LDH活性和心肌细胞凋亡指数降低(P<0.05);与NGFPc组比较,NGFPt组LVDP、HR和+ dp/dtmax升高,冠脉流出液LDH活性降低,心肌细胞凋亡指数降低(P<0.05),LVEDP和冠脉流出液CK-MB活性差异无统计学意义(P>0.05).I/R组心肌病理学损伤明显,而NGFPt组和NGFPC组明显减轻.结论 NGF-β(0.1 μg/ml)预先给药可减轻大鼠离体心脏缺血再灌注损伤,其效果优于预处理法.
Objective To evaluate the effect of pretreatment with nerve growth factor-beta (NGF-β) on ischemia-reperfusion (I/R) injury in isolated rat hearts by comparing it with NGF-β preconditioning.Methods Pathogen-free male Sprague-Dawley rats,weighing 200-300 g,aged 8-10 weeks,were anesthetized with intraperitoneal 10% chloral hydrate 300 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with KH solution aerated with 95% O2 and 5% CO2 at 37℃.Twenty-four isolated rat hearts were randomly divided into 3 groups (n =8 each) using a random number table:I/R group,NGF-β pretreatment group (group NGFPt) and NGF-β preconditioning group (group NGFPc).The hearts were perfused with K-H solution for 10 min (equilibration) in each group.In group I/R,the hearts were continuously perfused with K-H solution for 30 min.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min before ischemia in group NGF-Pt.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min followed by 10 min washout before ischemia in group NGFPc.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in each group.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 10 min equilibration (baseline,T1),immediately before suspension of perfusion (T2),and at 5,30,60 and 120 min of reperfusion (T3-6).The activities of creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) in coronary effluent were measured at T1 and T3-6.Myocardial specimens were obtained at T6 for detection of myocardial apoptosis (by TUNEL) and for microscopic examination.Apoptotic index (AI) was calculated.Results Compared with the baseline value at T1,+ dp/dtmax was significantly decreased,and LVEDP and activities of CK-MB and LDH were increased at T3-6 in each group,LVDP and HR were decreased at T3-6 in group I/R,LVDP was decreased at T3,4 in group NGFPt and at T3-6 in group NGFPc,and HR was increased at T2-6 in NGFPt and NGF Pc groups (P 〈 0.05).Compared with group I/R,LVDP,+ dp/dtmax and HR were significantly increased and LVEDP and activities of CK-MB and LDH and AI were decreased in NGFPt and NGFPc groups (P 〈 0.05).Compared with group NGFPc,LVDP,+ dp/dtmax and HR were significantly increased,while the LDH activity and AI were decreased (P 〈 0.05) and no significant changes were found in LVEDP and CK-MB activity in group NGFPt (P 〉 0.05).The pathologic changes of myocardium were significantly reduced in NGFPt and NGFPc groups as compared with I/R group.Conclusion Pretreatment with 0.1 μg/ml NGF-β attenuates I/R injury in isloated rat hearts,and the efficacy is superior to that of NGF-β preconditioning. Keywords:Nerve growth factor; Ischemic preconditioning; Myocardial reperfusion injury; Pretreatment
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2013年第9期1062-1065,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金青年基金(81000066)
关键词
神经生长因子
缺血预处理
心肌再灌注损伤
预先给药
Nerve growth factor
Ischemic preconditioning
Myocardial reperfusion injury
Pretreatment