加大科研管理力度 促进科技成果获奖——附1994—2004年我院科技成果获奖情况分析

伴随着改革开放的步伐，华中科技大学同济医学院附属协和医院（以下简称协和医院）已逐步建设成为拥有博士、硕士、学士学位授予权的具有较完备学科体系的部属“三甲一医院。肩负着教学、科研、医疗三重任务。科研成果的多少、质量的高低是衡量教学质量．医疗水平的重要标志，也是我院科研实力的具体表现。本丈是对我院1994～2004年获奖科技成果的情况进行一些回顾，从我院科研工作发展的轨迹中，客观地评价科研工作的优势和存在的不足，进而提出相应的对策，以提高我院的科技成果质量、探讨．提高成果获奖率的有效途径。     

热休克蛋白70对缺血再灌注未成熟心肌间质的保护作用

目的观察热休克蛋白70在未成熟心肌中的表达及对缺血再灌注未成熟心肌间质的影响。方法①实验于2001-12/2002-06在华中科技大学同济医学院药理实验室完成。选用出生14～21d的健康新生长耳大白兔12只。随机将兔分为2组对照组和实验组,每组6只。②对照组腹腔注射生理盐水0.4m L,注射后24h取体外心脏,常规建立Langendorff体外心脏灌注模型,灌注15m in转为工作心15m in后停灌45m in,恢复灌注15m in改为工作心30m in;实验组腹腔注射重酒石酸去甲肾上腺素,24h后取体外心脏,方法同对照组。③采用蛋白印迹法测定心肌细胞中热休克蛋白70含量。按公式计算羟脯氨酸含量(样本A值/标准A值×10×稀释倍数×1/组织质量)。采用均相放射免疫竞争法直接测定血浆内皮素1水平。④计量资料差异比较采用t检验。结果兔12只均进入结果分析。①实验组心肌细胞中热休克蛋白70含量(A值)明显高于对照组(P<0.01)。②实验组兔心肌羟脯氨酸含量明显高于对照组(P<0.01),血清内皮素1水平明显低于对照组(P<0.01)。结论腹腔注射重酒石酸去甲肾上腺素诱导未成熟大白兔心脏热休克蛋白70表达,在注射24h后热休克蛋白70表达明显增多,热休克蛋白70可明显减轻未成熟心肌内皮素的释放和心肌胶原的降解。     

血清HSP70与心肌缺血再灌注损伤的相关性研究

20只雄性Wister大鼠随机分为A(假手术)、B(缺血再灌注)两组,各10只,检测两组术前及术后1、3、5、12 h血清热休克蛋白70(HSP70)、肌酸激酶(CK)活性和心肌特异性肌钙蛋白(cTnT).结果显示,B组再灌注1、3、5、12 h血清HSP70水平较A组显著升高(P<0.05),并与CK及cTnT呈正相关(r分别为0.864、0.911,P均<0.01).认为缺血再灌注过程中大鼠血清HSP70水平升高,其升高程度反映了心肌缺血再灌注损伤程度.     

脂氧素A4对心肌缺血再灌注损伤的保护作用

Objective To investigate the roles of lipoxin A4, an endegenous lipid mediator with wide anti-inflammatory fea- tures, in attenuating myocardial ischemia-reperfusion injury and the possible mechanisms. Methods Thirty male KM mice were divid- ed randomly into three groups, 10 in each: ischemia-reperfusion group (group A), lipoxin A4 (0.1 mg/kg) group (group B) and Zn- PP (Zinc protoporphyrin Ⅸ, 25 mg/kg)phus lipoxin A4 (0.1 mg/kg)group (group C). A ischemia-reperfusion heart model was de- veloped by ligating the lift anterior descending branch of the coronary artery. A dine of 10% dehydrated alcohol in 0.2 ml for group A, a dose of isochoric lipoxin A4 for group B and a dose of isochoric ZnPP + lipoxin A4 for group C was infused into the ascending aorta through a catheter, which was kpassing the right common carotid artery, 30 minutes after reperfusion. The concentration of serum TNF-α, activities of serum crestine kinase(CK) and lactate dehydrogenase (LDH), activities of myeloperoxidase (MPO) and malond- ialdehyde (MDA) and the cell apoptosis rate in the myocardial tissue were measured 5 hours after reperfusion. Pathological features of the inflammatory infiltration in the myocardium were also observated.Results As compared with group A, the inflammatoryry infiltra- tion in the ischemic and necrotic regions tithe myocardium was reduced, with group C in the intermediate range. The serum activities of CK and LDH wine significantly lower in group B and C than that in group A, and the lowest activities were detected in group B. Similar findings were observed for MPO, an indicator for neutrophil infiltration, and MDA, an indictor for cell injury caused by oxy- gen radicals, in the myocardium. The concentration of TNF-α and the rate of cadiocyte apoptosis were decreased significantly in group B(P < 0.01). ZnPP, an inhibitor of heine oxygenase (HO)-1, attenusted the above protective effects of lipoxin A4 significantly (P<0.05). Conclusion Lipoxin A4 has protective effects against myocardial ischemia-reperfusion injury, and HO-1 may have a potential role in the protectve mechanisms of lipoxin A4, probably pertly by means of reducing the production of reactive oxygen spe- cies and TNF-α, decreasing the activation and infiltration of neutrophils, alleviating inflammatory damage and avoiding apoptosis.     

不同缺血预处理对未成熟心肌细胞功能的影响

目的比较不同方法的缺血预处理对未成熟心肌细胞功能的影响。方法采用Langendorff离体心脏灌注模型。分为4组:缺血/再灌注(I/R)组,离体心脏灌注15min转为工作心15min后停灌45min,恢复灌注15min改为工作心30min;心脏缺血预处理(MIP)组:离体心脏灌注15min转为工作心15min后反复2次缺血5min/再灌注5min,然后重复I/R组缺血/再灌注方法;肾缺血预处理(RIP)组:反复三次阻断左肾动脉5min,放开5min,切取心脏,重复I/R组方法;双下肢缺血预处理(DLIP)组:反复3次捆扎双下肢5min,松开5min,切取心脏,重复I/R组方法。以血清肌酸激酶(CK)和乳酸脱氢酶(LDH)漏出率、心肌组织三磷腺苷(ATP)和丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、心肌细胞内Ca2+含量、心肌线粒体Ca2+-ATPase活性及其Ca2+含量、心肌线粒体合成三磷腺苷能力[ATP]m作为观察指标。结果MIP、RIP及DLIP组ATP含量、SOD活性、心肌线粒体Ca2+-ATPase活性、[ATP]m均高于I/R组(P<0.01),MDA含量、CK、LDH漏出率、心肌细胞内Ca2+含量、心肌线粒体Ca2+含量均低于I/R组(P<0.01)。结论肾缺血预处理、双下肢缺血预处理与心脏缺血预处理有同等的心肌细胞保护作用。     

脂氧素A4对心肌缺血再灌注损伤的保护作用

Objective To investigate the roles of lipoxin A4, an endegenous lipid mediator with wide anti-inflammatory fea- tures, in attenuating myocardial ischemia-reperfusion injury and the possible mechanisms. Methods Thirty male KM mice were divid- ed randomly into three groups, 10 in each: ischemia-reperfusion group (group A), lipoxin A4 (0.1 mg/kg) group (group B) and Zn- PP (Zinc protoporphyrin Ⅸ, 25 mg/kg)phus lipoxin A4 (0.1 mg/kg)group (group C). A ischemia-reperfusion heart model was de- veloped by ligating the lift anterior descending branch of the coronary artery. A dine of 10% dehydrated alcohol in 0.2 ml for group A, a dose of isochoric lipoxin A4 for group B and a dose of isochoric ZnPP + lipoxin A4 for group C was infused into the ascending aorta through a catheter, which was kpassing the right common carotid artery, 30 minutes after reperfusion. The concentration of serum TNF-α, activities of serum crestine kinase(CK) and lactate dehydrogenase (LDH), activities of myeloperoxidase (MPO) and malond- ialdehyde (MDA) and the cell apoptosis rate in the myocardial tissue were measured 5 hours after reperfusion. Pathological features of the inflammatory infiltration in the myocardium were also observated.Results As compared with group A, the inflammatoryry infiltra- tion in the ischemic and necrotic regions tithe myocardium was reduced, with group C in the intermediate range. The serum activities of CK and LDH wine significantly lower in group B and C than that in group A, and the lowest activities were detected in group B. Similar findings were observed for MPO, an indicator for neutrophil infiltration, and MDA, an indictor for cell injury caused by oxy- gen radicals, in the myocardium. The concentration of TNF-α and the rate of cadiocyte apoptosis were decreased significantly in group B(P < 0.01). ZnPP, an inhibitor of heine oxygenase (HO)-1, attenusted the above protective effects of lipoxin A4 significantly (P<0.05). Conclusion Lipoxin A4 has protective effects against myocardial ischemia-reperfusion injury, and HO-1 may have a potential role in the protectve mechanisms of lipoxin A4, probably pertly by means of reducing the production of reactive oxygen spe- cies and TNF-α, decreasing the activation and infiltration of neutrophils, alleviating inflammatory damage and avoiding apoptosis.     

金属硫蛋白对未成熟心肌和心肌间质的保护作用

目的探讨诱导金属硫蛋白(MT)在未成熟心肌中的表达对缺血-再灌注未成熟心肌和心肌间质的影响。方法将24只兔(14~21d)按随机数字表法分为4组,每组6只,对照组:腹腔注射蒸馏水0.3ml,24h后取离体心脏行Langendorff灌注;组1、组2和组3在腹腔注射3.6%ZnSO4(1.5ml/kg),分别于注射后12h、24h和48h取离体心脏行Langendorff灌注。测定心肌细胞中MT含量、血流动力学、生化指标,观察心肌超微结构改变。结果组2、组3与对照组、组1比较,MT含量、三磷酸腺苷(ATP)含量、超氧化物歧化酶活性、羟脯氨酸含量、心肌线粒体[Ca2+-ATPase]m、心肌线粒体合成ATP的能力明显增高(P<0.01),心肌含水量、丙二醛含量、心肌肌酸激酶、乳酸脱氢酶漏出率、心肌线粒体Ca2+含量、心肌线粒体[Ca2+]m和内皮素含量明显降低(P<0.01),心肌超微结构损伤明显减轻。结论腹腔注射ZnSO4可诱导心肌MT长时间表达,MT可减轻未成熟心肌和心肌间质的缺血-再灌注损伤。     

血管内皮细胞生长因子在大鼠心脏移植急性排斥期的表达研究

目的:观察血管内皮细胞生长因子(VEGF)在大鼠心脏移植急性排斥期中的表达及与排斥反应的关系。方法:实验分为对照组和环孢霉素A(CSA)组,每组27只。采用颈部心脏异位移植术式建立移植模型,于心脏移植手术后分别静脉给予0.85%氯化钠溶液及CSA干预。常规监测排斥反应发生情况。每组7只用于观察移植物存活时间,余20只移植术后1,3,7,11d各切取5例移植心标本。样本采用逆转录聚合酶链反应(RTPCR)的方法检测移植心VEGF的DNA表达水平。结果:CSA组移植心存活时间[(21.1±2.9)d]长于对照组[(12.4±2.3)d],P<0.01;对照组各采样时段VEGF的DNA表达强度均强于CSA组,P<0.01。结论:VEGF高表达与移植心的炎性浸润以及急性排斥有密切关系。     

热休克蛋白70对兔未成熟心肌和心肌间质的影响

目的: 探讨热休克蛋白70(HSP70)对未成熟心肌和心肌间质的影响. 方法:健康新生长耳大白兔12只随机分为2组. 对照组(C组):ip生理盐水0.4 mL,24 h后取离体心脏,常规建立Langendorff离体心脏灌注模型,灌注15 min转为工作心15 min后停灌45 min,恢复灌注15 min改为工作心30 min;实验组(E组):ip去甲肾上腺素, 24 h后取离体心脏,方法同对照组. 测定心肌细胞中HSP70含量、血流动力学指标、心肌含水量(MWC)、心肌肌酸激酶(CK)和乳酸脱氢酶(LDH)漏出率、三磷酸腺苷(ATP)含量、超氧化物歧化酶(SOD)和丙二醛(MDA)含量、心肌组织羟脯氨酸(HP)含量、内皮素(ET) 含量、心肌细胞内Ca2 含量、心肌线粒体Ca2 -ATPase活性及其Ca2 含量、心肌线粒体合成ATP能力[ATP]m,心肌超微结构. 结果:E组HSP70含量明显高于C组(P<0.01);MWC低于C组(P<0.05);ATP含量、SOD活性、心肌线粒体Ca2 -ATPase活性、[ATP]m, HP含量优于C组(P<0.01),MDA含量、CK, LDH漏出率、心肌细胞内Ca2 含量、心肌线粒体Ca2 含量、ET含量低于C组(P<0.01),心肌超微结构损伤较C组明显减轻. 结论:HSP70对缺血再灌注未成熟心肌和心肌间质具有明显的保护作用.