Proteomic profiling of dog urine after European adder (Vipera berus berus) envenomation by two-dimensional difference gel electrophoresis

Between April and September every year, many dogs in Finland are bitten by Vipera berus berus, also known as the European adder, the only venomous snake in the area. Exposure to snake bite venom causes local and systemic symptoms and in severe cases can lead to death. Urine samples were collected from four dogs bitten by V.berus berus and treated in the intensive care unit of the Veterinary Teaching Hospital at the University of Helsinki. The inclusion criteria were a strong suspicion of an adder bite no more than two days before admission and clinical signs of an adder bite. Exclusion criteria were defined as ongoing treatment with glucocorticoids or a known history of liver or kidney diseases. Six privately owned, healthy dogs were obtained as controls. Samples were subjected to 2D-DIGE analysis. Image analysis was performed with DeCyder 7.0 2D software, and protein spots demonstrating a minimum 1.5-fold difference in average spot volume ratios between envenomed and control dogs with a Student's t-test p-value of less than 0.05 were picked and identified using LC-MS/MS. In 2D-DIGE analysis, seven proteins were significantly (p < 0.05) over-expressed in the urine of dogs bitten by V.berus berus compared to the control group. From these, five proteins were identified: beta-2-microglobulin (b2MG), alpha-1-antitrypsin (AAT), albumin, fetuin-B and superoxide dismutase (SOD1). Results indicate that envenomation by V.berus berus alter the urinary protein profile in dogs.     

乌司他丁对颅内血管疾病手术SOD、MDA的影响

目的探讨乌司他丁在颅内血管手术围术期脑保护的作用。方法择期行颅内血管手术成年患者30-N,随机分为鸟司他丁组（U组）和对照组（C组）各15例。麻醉诱导前,u组5min内静注乌司他丁5000IU／kg（用0．9％生理盐水20ml稀释）;C组同速度输入等量生理盐水。分别于麻醉诱导前（Tn）、手术lh（T,）、手术2h（T2）、手术6h（T。）抽取颈静脉球部血测SOD、MDA。记录两组患者手术后24h格拉斯哥GCS评分以及术后恢复情况。结果U组T：、T,的SOD、MDA较L差异无统计学意义（P〉0．05）;C组T,、T。的SOD较]rn降低,T2、L的MDA较T0升高（P〈0．05）。组间比较u组T2、T3的SOD、MDA较c组差异有统计学意义（P〈0．05）。两组术后24h的GCS比较无明显差异（P〉0．05）。结论乌司他丁预处理对颅内血管疾病手术具有脑保护的作用。     

Effect of some free radical scavengers on reperfusion induced arrhythmias in the isolated rat heart

The possible role of oxygen free radicals in the development of reperfusion arrhythmias was investigated using a 10-min period of coronary ligation followed by reperfusion in the isolated rat heart. Superoxide dismutase (5 to 20 u/ml) glutathione (10(-5) to 10(3)M) and ascorbic acid (10(-4) to 5 X 10(-4) M) when given before coronary ligation attenuated the development of reperfusion arrhythmias. Mannitol (2 X 10(-2)M) and catalase (100 and 300 u/ml) did not have any significant effect on reperfusion arrhythmias when given alone but they did potentiate the antiarrhythmic effect of superoxide dismutase. Glutathione, and a combination of superoxide dismutase, catalase and mannitol also reduced the incidence of reperfusion induced ventricular fibrillation when given just before reperfusion. By perfusing hearts with ferricytochrome C it was possible to show an increased reduction of ferricytochrome C during the first minute of reperfusion which could be prevented by the addition of superoxide dismutase. These results provide evidence that oxygen free radicals are produced and may be important in the genesis of reperfusion induced arrhythmias in the isolated rat heart.     

人基因工程重组超氧化物歧化酶对骨髓造血干细胞的辐射防护作用

用照射小鼠的骨髓造血干细胞(CFU-S)外源性脾结节形成法,评价人基因工程重组SOD(rhSOD),聚乙二醇(PEG)修饰的长半衰期重组SOD(PEG-SOD)对CFU-S的辐射防护作用.结果显示:rhSOD和PEG-SOD均能明显提高CFU-S产率,其中rh SOD在照射前1h给药效果好,而PEG-SOD在照射前2～3h给药作用更为明显,化学修饰后SOD血浆半衰期延长,将使给药时机及药物作用持续时间得到改善.     

长效重组人超氧化物歧化酶的研制及其辐射防护作用

选用无毒、无免疫原性的高分子化合物聚乙二醇(PEG)为修饰剂,以活性酯法制备出PEG- 重组人超氧化物歧化酶(PEG-rh SOD)共价结合物.修饰酶的生物半衰期为15h,比修饰前延长了90倍,对酸、碱、热的耐受力明显高于修饰前.用照射小鼠的活存率评价了PEG-rhSOD的辐射防护作用.小鼠受8.5Gy γ射线照射前1h腹腔注射30万U/kg PEG-rh SOD,可提高活存率42%,与未修饰的rhSOD相比,活存率提高14%.实验仍在进行中.     

Role of reactive oxygen in phospholipase A2 activation by ischemia/reperfusion of the rat kidney

Purpose. To investigate the role of phospholipase A₂ (PLA₂) in reperfusion injury of the kidney in an in vivo animal model, renal mitochondrial PLA₂ activity was measured under three different conditions. Methods. Male Wistar rats (n = 72) anesthetized with pentobarbital underwent renal ischemia surgically for 45 min and were reperfused for the indicated time (renal ischemia/reperfusion). Treatments included reperfusion for various predetermined periods (phase 1), exposure to hyperbaric oxygen (phase 2), and administration of reactive oxygen species (ROS) scavenger (phase 3). Thereafter, each kidney was harvested, and mitochondrial PLA₂ activity was measured by a radioisotope technique. Results. Ischemia/reperfusion resulted in time-related PLA₂ activation in the renal mitochondria up to 48 h of reperfusion after renal ischemia. Renal mitochondrial PLA₂ activity was further augmented by hyperbaric oxygen exposure prior to reperfusion, whereas administration of the ROS scavengers suppressed mitochondrial PLA₂ activity. Conclusion. These data suggest that ROS may play an important role in the in vivo activation of PLA₂ associated with renal ischemia/reperfusion. Received for publication on July 6, 1998; accepted on November 30, 1998     

异丙酚对内毒素血症鼠NO、SOD及MDA水平的影响

目的 研究异丙酚对内毒素血症大鼠体内一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)的影响。方法 72只Waster雄性大鼠随机分为三组,对照组(C组),内毒素组(L组)和异丙酚 内毒素组(P组)。L组腹腔内注入内毒素10mg·kg-1;P组皮下缓慢注入异丙酚20mg·kg-1·h-1后再注入内毒素10mg·kg-1;C组:腹腔内注入等量生理盐水。L组和P组在注射后30、90、180、360min取静脉血并处死动物(每时间点8只),C组在相应时间点留取3ml静脉血后,于360min处死大鼠留取肺组织。测定各时间点血清和肺组织中MDA和NO浓度及外周红细胞SOD活性。结果 P组用异丙酚后显著性降低内毒素血症大鼠血清和肺组织中NO和MDA浓度,并增加SOD活性,与L组比较差异有显著性(P<0.01或P<0.05)。结论 异丙酚可降低内毒素血症鼠NO水平,抑制MDA合成,增加SOD活性,具有一定的抗氧化作用,可改善内毒素血症鼠的功能状态。     

益心康泰胶囊对老年SD大鼠生精功能及血清超氧化物歧化酶和丙二醛的影响

目的:观察益心康泰对老龄SD大鼠精子密度和活率,以及血清超氧化物歧化酶(SOD)和丙二醛(MDA)的影响,并探讨其可能的作用机制。方法:将老年(24月龄)SD雄性大鼠随机分为空白组、十一酸睾酮组、益心康泰高、中、低剂量组共5组,根据体重,分别给予10 ml/kg生理盐水、4 mg/kg十一酸睾酮悬浊液、0.6 g/kg、0.3 g/kg、0.15 g/kg益心康泰悬浊液灌胃,1次/d,30 d后,取附睾,剥离脂肪、包膜,剪碎放入37℃恒温水浴锅中预热的生理盐水试管内,利用CASA精子自动分析系统(伟力)检测精子各项常规参数。取血查血清SOD和MDA含量。并行睾丸组织HE染色,在光镜下观察睾丸组织形态学变化。结果:与空白对照组相比,高、中、低剂量益心康泰均能显著提高老年大鼠的精子密度[分别为(152.3±24.0)×106/ml、(334.5±97.7)×106/ml、(302.7±158.9)×106/ml、(221.4±75.6)×106/ml,P<0.05]和活率[分别为(26.5±2.0)%、(35.5±5.9)%、(33.1±5.4)%、(28.4±2.1)%,高、中剂量与空白组相比P<0.01],同时,高剂量组可以显著提高血清SOD含量[(22.0±5.3)U/ml,P<0.01],降低MDA含量[(13.6±4.6)nmol/ml,P<0.01];睾丸组织学检测显示,给药组相对于空白组细胞增殖更为活跃,可见较多分裂期细胞,管腔内成熟精子数目较多,间质稀少,血管丰富,间质细胞发育良好。结论:益心康泰可以通过影响SOD和MDA的含量而改善老年SD大鼠的生精功能。     

胺碘酮联合磷酸肌酸钠对窒息兔心肺复苏后心肌形态学及丙二醛、超氧化物歧化酶的影响

目的 探讨胺碘酮联合磷酸肌酸钠对窒息兔心肺复苏( cardiopulmonary resuscitation,CPR)后心肌形态学、丙二醛( malondialdehyde,MDA)含量及超氧化物歧化酶(superoxide dismutase,SOD)活性的影响.方法 采用窒息法制作CPR模型.选取健康日本长耳大白兔40只,完全随机分为A组(假手术组)、B组(生理盐水组)、C组(胺碘酮5 mg/kg组)、D组(磷酸肌酸钠100 mg/kg组)、E组(胺碘酮5 mg/kg联合磷酸肌酸钠100 mg/kg组),每组8只.CPR后6h处死兔,光镜及电镜下观察心肌形态学变化,检测心肌MDA含量、SOD活性.结果 光镜下心肌组织病理改变显示:E组心肌损伤轻于C组和D组,C组和D组又轻于B组.电镜下心肌超微结构显示:A组兔的心肌线粒体完整,肌丝排列整齐,明暗带清晰;B组兔的心肌线粒体部分水肿、空泡化,线粒体嵴断裂,肌丝排列紊乱,明暗带分界不清;C组兔心肌少数线粒体空泡化、嵴断裂,肌丝排列尚整齐,明暗带显示不清;D组兔心肌少数线粒体空泡化、嵴断裂,明暗带清晰;E组兔心肌线粒体完整,个别线粒体水肿、嵴断裂,明暗带清晰可见.B组、C组、D组兔心肌MDA含量(2.45±0.41,1.65±0.18,1.71±0.17)较A组(0.98±0.22)升高,差异有统计学意义(P＜0.05);B组兔心肌MDA含量显著高于C组和D组,C组和D组又高于E组,差异有统计学意义(P＜0.05);E组(1.21±0.55)与A组比较差异无统计学意义.B组、C组、D组和E组兔心肌SOD活性(16.2±0.9,17.9±1.0,18.2±0.6,19.8±1.4)较A组(21.4±2.1)减低,差异有统计学意义(P＜0.05);B组心肌SOD活性低于C组和D组,C组和D组又低于E组,差异有统计学意义(P＜0.05).结论 胺碘酮联合磷酸肌酸钠能减轻CPR后心肌损伤;部分抑制CPR后的过氧化反应,减轻脂质过氧化损伤,可能是胺碘酮联合磷酸肌酸钠心肌保护作用的主要机制之一.     

兔肝癌组织缺血再灌注后的损伤及凋亡

目的研究肝癌缺血再灌注后的损伤及凋亡。方法超声引导穿刺新西兰兔肝脏左中叶注射VX2肿瘤组织混悬液,建立肝脏肿瘤模型,阻断肿瘤所在的肝左中叶的肝动脉分支60min后去除血管阻断恢复血流,分别再灌注0min、1h、1d、3d和1w。取肿瘤组织、癌周组织和肝脏组织,除测定肿瘤组织和肝脏组织超氧化物歧化酶(SOD)、丙二醛(MDA)的含量外,还对肿瘤组织、癌周组织及肝脏组织进行了HE染色。结果缺血再灌注后肿瘤组织和正常肝组织中的SOD浓度均迅速下降并分别于再灌注1h和0min达最低水平(64.59±4.97,12.38±0.31),其后逐渐恢复(112.83±5.84,25.78±0.56)但至再灌注7d仍低于再灌注前(200.32±26.43,42.00±1.07)。但癌组织中的SOD含量下降更为显著。从缺血再灌注开始至7d肿瘤组织的MDA含量下降(21.59±0.59),而肝脏组织缺血再灌注后0min至7d(29.04±1.43),MDA含量明显升高,均高于缺血再灌注前水平(18.26±0.43)。HE染色显示,肝癌组织缺血再灌注后凋亡细胞明显增多,其中以1d最为显著(23.08%),而癌周和肝组织改变不甚明显(与同时点的癌组织相比P<0.01)。结论缺血再灌注可增强对肝癌组织的损伤并促进细胞凋亡而癌周和正常肝脏组织的改变不明显。