ghrelin对失血性休克大鼠胃黏膜的保护作用

目的 探讨重组大鼠ghrelin对失血性休克大鼠胃黏膜损伤的影响.方法 18只健康雄性SD大鼠被随机分为对照组、失血性休克组、ghrelin治疗组3组.每组6只.失血性休克组及ghrelin治疗组制备失血性休克模型.ghrelin治疗组休克复苏的同时经颈内静脉推注重组大鼠ghrelin 20μg/kg.于复苏后2 h使用激光多普勒血流仪测定大鼠胃黏膜血流量;用光镜及透射电镜观察胃黏膜损伤情况.结果 失血性休克组胃黏膜血流量较对照组明显降低(260.4±49.6)bpu比(418.6±57.3)bpu,P<0.013,胃黏膜细胞坏死、脱落、溶解,局部溃疡形成.ghrelin治疗组胃黏膜血流量较失血性休克组明显提高((352.9±72.9)bpu比(260.45±49.6)bpu,P<0.05),且与对照组比较差异无统计学意义(P>0.05);胃黏膜损伤显著改善.结论 重组大鼠ghrelin能够提高失血性休克大鼠胃黏膜血流量,改善胃黏膜缺血/再灌注损伤.     

高压氧对大鼠肠缺血-再灌注损伤致细胞凋亡的影响

目的 观察大鼠在小肠缺血-再灌注(I/R)不同时期应用高压氧(HBO)治疗对小肠黏膜细胞凋亡的影响,并探讨其作用机制.方法 采取大鼠肠系膜上动脉(SMA)钳夹缺血60 min,松钳夹再灌注60 min,建立S-D大鼠小肠I/R损伤模型,并随机(随机数字法)分成4组:I/R组、缺血前HBO治疗组或HBO预处理组(HBO-P)、缺血期HBO治疗组(HBO-I)和再灌注期HBO治疗组(HBO-R).再灌注60 min后,取回肠末端小肠标本.采用酶连免疫吸附试验法(Elisa)检测肠道组织TNF-α,用比色法检测肠道组织匀浆含量ATP,用免疫组化法检测小肠组织中半胱氨酸天门冬氨酸特异蛋白酶(Caspase-3)的表达.数据以均数±标准差(-x±s)表示,采用单因素方差分析检验,独立样本间比较采用SNK-q检验.结果 肠道组织TNF-α含量在HBO-I组最低,其次为HBO-P组(每两组比较P＜0.05),但前两组明显低于HBO-R组和I/R组(P＜0.05),HBO-R组略低于I/R组,但差异无统计学意义(P＞0.05);Caspase-3表达在HBO-I组最低,HBO-P组次之(两组比较P＜0.05);HBO-R和I/R组最高(与前两组比较P＜0.05),尽管HBO-R组略低于I/R组,但差异无统计学意义(P＞0.05);ATP含量在HBO-I组最低,HBO-P组次之(两组比较P＜0.05),HBO-R和I/R组最高(与前两组比较P＜0.05),尽管HBO-R组略低于I/R组,但差异无统计学意义(P＞0.05).结论 HBO、小肠I/R损伤和小肠黏膜上皮凋亡之间存在联系;HBO治疗可以维持黏膜上皮细胞能量代谢,减少ATP耗竭,降低肠道组织TNF-α含量,减轻I/R损伤小肠黏膜上皮凋亡;在缺血期和缺血前应用HBO治疗显示有益结果,特别是在缺血期应用HBO效果最好,再灌注期应用HBO无效.     

盐酸戊乙奎醚结合液体复苏抑制失血性休克大鼠小肠黏膜凋亡的研究

目的 探讨盐酸戊乙奎醚结合液体复苏对失血性休克大鼠小肠黏膜细胞凋亡的影响.方法 32只SD大鼠,制成重度失血性休克模型,分成对照组、假手术组、液体复苏组和盐酸戊乙奎醚结合液体复苏组,每组8只.采用流式细胞仪和FITC-Annexin V/PI荧光染色法检测和比较失血/复苏/急救后各组大鼠小肠黏膜细胞凋亡的发生情况.结果 无论采取治疗措施,经历了失血性休克和复苏的大鼠,其小肠黏膜细胞均存在显著的细胞凋亡情况.液体复苏组和盐酸戊乙奎醚结合液体复苏组大鼠的小肠黏膜细胞凋亡率明显高于对照组和假手术组,差异均有统计学意义（P均＜0.05）.同时,与液体复苏组相比较,盐酸戊乙奎醚结合液体复苏组大鼠的小肠黏膜细胞凋亡率明显降低,差异有统计学意义（P＜0.05）. 结论 盐酸戊乙奎醚结合液体复苏能有效抑制失血性休克大鼠肠道黏膜细胞的凋亡,从而减少失血/复苏对肠道黏膜完整性的损害,可能有助于改善预后.     

早期胃内灌注ω-3多不饱和脂肪酸对失血性休克大鼠肠黏膜屏障功能影响的实验研究

[目的]探讨失血性休克复苏后早期胃内灌注ω-3多不饱和脂肪酸（ω-3 PUFAs）对大鼠肠黏膜屏障功能的影响.[方法]选取健康成年SD大鼠72只,按照胃内灌注状况,随机分为ω-3 PUFAs组、牛奶组、不干预组.乌拉坦腹腔麻醉后,制作休克大鼠模型,休克30 min后复苏,ω-3PUFAs组和牛奶组在复苏结束、复苏2h分别进行胃内灌注.各组分别在休克30 min、复苏2h和4h处死1/3大鼠,取肝脏组织进行细菌培养,取一段空肠制成病理切片.[结果]肝脏组织细菌培养显示,ω-3 PUFAs组、牛奶组复苏2h、4h菌落数较休克30min少,同时限点也较不干预组少（P＜0.05）;肠道病理切片显示,复苏2h、4hω-3 PUFAs组较牛奶组和不干预组轻,不干预组损伤最重.[结论]失血性休克救护过程中,早期胃内灌注ω-3PUFAs更有利于肠道屏障功能的恢复、减少肠内细菌移位.     

大鼠失血性休克复苏早期肠黏膜损伤与修复的形态学观察

目的观察失血性休克复苏后早期，大鼠肠黏膜损伤修复的形态学特征。方法建立失血性休克致肠缺血-再灌注模型，通过光镜和电镜观察不同阶段肠黏膜的改变及肠上皮损伤指数。结果从复苏0h起回肠和空肠肠道上皮就发生明显的损伤改变，至3h损伤改变仍然存在，以1h最为明显；3h起部分肠黏膜即出现修复现象，6h大部分黏膜已修复，24h肠黏膜结构恢复正常；大肠与小肠在损伤和修复的变化过程方面基本相同，但其程度明显低于小肠；小肠黏膜厚度和绒毛高度1h后逐渐减少，大肠各组间则无明显变化。结论失血性休克致肠缺血-再灌注损伤早期肠黏膜屏障受累，同时损伤的肠黏膜能够得到快速修复重建，大肠比小肠具有更强的抗损伤能力。     

大鼠肠缺血再灌注损伤的实验研究

目的:通过大鼠失血性休克模型,探讨肠缺血再灌注损伤后肠黏膜的损伤情况及肠屏障功能的变化。方法:SD大鼠40只,分对照组(假休克)和休克组,后者又分为休克复苏后1h、3h和6h3小组,每小组大鼠10只。颈动脉和颈静脉插管,通过放血使大鼠的平均动脉压降至40mmHg,持续70min后回输血和生理盐水复苏,血压平稳后分别以各时点取血和回肠标本进行组织学、肠黏膜损伤度检查和D-乳酸(D-LAC)、二胺氧化酶(DAO)活性测定。结果:大鼠失血性休克后,肠黏膜明显受损,组织学检查主要为黏膜的水肿、灶性坏死、绒毛脱落及炎性细胞浸润,复苏后1h、3h、6h肠黏膜损伤指数分别达3.0、2.4、1.6;外周血DAO活性升高显著(与对照组相比,1h、3h和6h组P值均<0.05);D-LAC于复苏后1h后显著升高(与对照组相比,P<0.05)。结论:大鼠失血性休克后肠黏膜明显损伤,肠通透性增加。     

胸腺五肽对重度创伤失血性休克大鼠SIgA的影响

目的 探讨早期加用胸腺五肽(TP-5)对重度创伤失血性休克大鼠SIgA的影响.方法 72只健康雄性SD大鼠随机分为常规复苏组和TP-5复苏组,每组36例.常规复苏组选用乳酸林格液复苏;TP-5复苏组在常规复苏的基础上早期加用TP-5.在休克前(t0)、休克复苏前(t1)及休克复苏或复苏给药后12 h(t2)、24 h(t3)、48 h(t4)、72 h(t5)各时间点采集肠道黏液,应用ELISA试剂盒测定SIgA含量,并在光学显微镜下进行小肠上皮损伤指数评分.结果 与休克前比较,复苏前后两组SIgA含量均明显降低(P<0.01),随复苏时间的延长,SIgA含量逐渐升高,两组复苏后12 h、24 h、48 h、72 h比较差异有统计学意义(P<0.05).两组休克复苏前及复苏后12 h、24 h、48 h小肠上皮损伤指数均明显升高(P<0.05,P<0.01),随复苏时间的延长,小肠上皮损伤指数逐渐回降,TP-5组复苏后24 h、48 h明显优于常规复苏组(P<0.05).结论 重度创伤失血性休克大鼠复苏早期联合应用TP-5较常规复苏治疗可显著提高SIgA的分泌量,加快小肠黏膜形态、结构完整性的恢复,保护肠道黏膜屏障,有益于提高机体免疫能力.     

脑缺血再灌注老龄大鼠脑组织核转录因子кB和一氧化氮合酶的变化

目的从脑组织核转录因子κB、热休克蛋白和一氧化氮合酶(eNOS、iN-OS、nNOS)表达变化揭示老年脑缺血再灌注损伤可能的病理生理机制.方法实验于2003-01/2004-09在河南中医学院老年医学研究所完成.SD雄性青年(五六月龄)、老龄(21月龄)大鼠各44只,采用大脑中动脉局灶性脑缺血法复制动物模型,排除手术过程中死亡和造模不成功者,设青年假手术组(n=6)、青年模型组(n=30)、老龄假手术组(n=6)、老龄模型组(n=30).各模型组又分为缺血3 h和再灌注1,3,6,12 d 4个时间点,每个时间点6只.观察缺血3 h和再灌注1,3,6,12 d神经症状积分、脑组织含水量、病理变化、核转录因子κB、热休克蛋白、一氧化氮合酶表达的变化.结果实验大鼠青年组36只、老龄假组36只,全部进入结果分析.①青年模型组和老龄模型组脑组织含水量(各时间点)、神经症状积分(再灌注1,3,6 d)、核转录因子κB(再灌注1,3,6,12 d)、热休克蛋白(缺血3 h,再灌注1,3,6 d)、eNOS(再灌注1,3,6 d)、nNOS(各时间点)和iNOS(再灌注1,3,6 d)的表达分别高于同时段青年、老龄假手术组.②老龄模型组神经症状积分(缺血3 h,再灌注6 d)、核转录因子κB(再灌注1,3 d)、nNOS(缺血3 h,再灌注1 d)和iNOS(再灌注1,3 d)表达高于青年模型组,而eNOS(再灌注3,6 d)和热休克蛋白(缺血3 h,再灌注1 d)表达低于同时段青年模型组.③各组大鼠脑组织病理观察,假手术大鼠大脑皮质各层神经元结构完整,无炎性浸润、出血灶等.青年模型组大鼠神经元、胶质细胞肿胀,血管扩张充血、管周水肿、炎性细胞浸润积聚.老龄模型组大鼠神经元、胶质细胞肿胀,排列紊乱,血管扩张充血、管周水肿、炎性细胞浸润(血管套),血管外细胞外间隙水肿.④脑组织神经细胞、微血管基底膜电镜观察,假手术组神经元细胞器形态结构正常.青年模型组大鼠脑组织神经元、胶质细胞造模3 h胞质、线粒体开始水肿,线粒体嵴和膜开始消失,内质网脱颗粒;微血管周围轻度水肿,部分基底膜断、水肿;1～3 d逐步加重,12 d损伤减轻.老龄模型组大鼠脑组织神经元、胶质细胞造模3 h后胞质、线粒体高度水肿,线粒体嵴和膜消失,微血管周围水肿,基底膜缺损;3 d超微结构破坏达到最严重,微血管基底膜大部分溶解、破坏,甚至消失,内皮细胞胞膜缺损,绒毛缺失.结论脑缺血再灌注损伤与核转录因子κB、iNOS、nNOS表达增强有关,老年脑缺血再灌注损伤严重可能为随着增龄核转录因子κB、iNOS、nNOS表达增强和热休克蛋白、eNOS表达降低所致.     

肝动脉/门静脉早期复流对肝移植大鼠小肠缺血再灌注损伤的影响

背景:当肝动脉与门静脉早期复流时序不同时,是否会加重对肝移植大鼠小肠缺血/再灌注的损伤尚未见大量报道.目的:探讨肝动脉与门静脉早期复流对肝移植太鼠小肠缺血/再灌注损伤的影响.方法:采用门静脉灌注的大鼠自体肝移植模型,78只SD大鼠以简单随机化法分为3组:肝动脉组(忙36):行自体肝移植手术,以40C乳酸林格液由门静脉灌肝40min,开放肝动脉及下腔静脉,10min后开放门静脉;门静脉组(n=36):行自体肝移植手术,门静脉开放恢复肝脏血流后10 min再开放肝动脉血流:假手术组(n=6):打开腹腔,游离肝脏后关腹.观察各组小肠显微及超微结构变化并测定一氧化氮水平.结果与结论:术后各实验组不同时段先后出现小肠绒毛排列不整或紊乱,小肠黏膜细胞线粒体大小不一,明显肿胀,呈类圆形,内有空泡变性,严重者可见嵴减少、断裂或消失.小肠组织一氧化氮水平均升高.上述变化在术后12 h达高峰.术后肝动脉先复流组小肠显微及超微结构损伤及小肠组织一氧化氮水平明显高于门静脉先复流组.提示,肝动脉早期复流可以通过早期肝脏供氧以减少移植肝脏的损害,但门静脉的延迟开放则加重了肝移植大鼠小肠的缺血,再灌注损伤.     

重组人促红细胞生成素后处理缺血/再灌注前后骨骼肌诱导型一氧化氮合酶及一氧化氮的变化

背景:促红细胞生成素对包括心脑在内的多种组织器官的缺血/再灌注损伤有保护作用.目的:观察重组人促红细胞生成素后处理对家兔后肢腓肠肌缺血/再灌注前后诱导型一氧化氮合酶、一氧化氮和超微结构的影响.方法:将家兔随机分为空白对照组、模型组和重组人促红细胞生成素组,后2组建立兔左后肢腓肠肌缺血/再灌注实验模型,其中重组人促红细胞生成素组在兔左后肢缺血2 h后静脉注射重组人促红细胞生成素.结果与结论:再灌注后4,12 h,重组人促红细胞生成素组诱导型一氧化氮合酶表达水平及一氧化氮浓度增高幅度较模型组低(P < 0.05).电镜下腓肠肌细胞超微结构显示,模型组内皮细胞膜溶解,肿胀明显,肌纤维内线粒体水肿.重组人促红细胞生成素组肌纤维损伤较轻,肌节内Z线及肌节内各带结构基本正常,大部分线粒体结构正常,显示重组人促红细胞生成素组超微结构损伤程度明显轻于模型组.结果证实,重组人促红细胞生成素后处理可通过抑制诱导型一氧化氮合酶蛋白的表达,减少再灌注后过量一氧化氮生成,改善骨骼肌缺血/再灌注损伤.     

失血性休克和内毒素二次打击肺损伤时转化生长因子-β1/smad2信号通路的变化

目的 观察失血性休克(HS)+内毒素二次打击致肺损伤时肺转化生长因子-β1(TGF-β1)/smad2信号通路的变化.方法 24只SD大鼠被随机分为假手术(Sham)组和HS组,每组12只.HS组建立未控制HS+内毒素二次打击模型,并按失血前期90 min、复苏期60 min、观察期210 min进行实验.监测各时间点平均动脉压(MAP)、心率、呼吸频率,实验结束后取血测定动脉血气,活杀大鼠取肺组织测定肺毛细血管通透性及肺湿/干重(W/D)比值,采用苏木素-伊红(HE)染色在光镜下观察肺组织病理学改变,用免疫组化法和逆转录-聚合酶链反应(RT-PCR)测定肺组织TGF-61的蛋白及mRNA表达,用蛋白质免疫印迹法(West-ern blotting)测定肺组织smad2蛋白表达.结果 与Sham组比较,HS组MAP自院前期60 min后明显降低,血乳酸(Lac)明显升高,pH值、动脉血氧分压(PaO2)、HCO-3、动脉血氧饱和度(SaO2)均明显降低,剩余碱(BE)负值增大,肺毛细血管通透性及W/D比值明显升高(P均＜0.01);HS组肺组织TGF-β1呈强阳性表达,主要分布于肺泡上皮、间质炎性细胞、肺泡腔内巨噬细胞和小支气管黏膜上皮细胞;TGF-β1 mRNA和smad2蛋白表达均较Sham组明显上调(P均＜0.01).结论 二次打击大鼠TGF-β1/smad2信号通路被激活,并与肺毛细血管通透性和肺损伤的严重程度密切相关,可能是HS后肺损伤的重要机制之一.     

Hypertonic saline improves intestinal mucosa barrier function and lung injury after trauma-hemorrhagic shock

Our objective was to test the hypotheses that small volume hypertonic saline (HTS) resuscitation protects against trauma-hemorrhagic shock (T/HS)-induced intestinal and lung injury better than standard volume resuscitation with Ringer's lactate (RL), and that the degree of lung injury correlates with the degree of gut injury after therapy. Male Sprague-Dawley rats were subjected to laparotomy (trauma) and 90 min of T/HS or sham shock (T/SS), and were then resuscitated with RL or 7.5% NaCl solution at an equivalent sodium load. Intestinal and lung injury was assessed at 3 and 24 h after resuscitation. Lung permeability, pulmonary myeloperoxidase (MPO) levels, and the bronchoalveolar lavage fluid (BALF) protein to plasma protein ratio were increased after T/HS, but were significantly lower in HTS-resuscitated than RL-treated rats. The incidence of bacterial translocation (BT) was not different between the groups, but the magnitude of BT after T/HS was less after HTS than RL resuscitation. Barrier function of intestinal segments was impaired only in the T/HS rats resuscitated with RL and histological analysis demonstrated fewer injured villi in the T/HS rats resuscitated with HTS than RL. Linear regression analysis revealed direct correlations between the percent of injured villi, increased lung permeability, and pulmonary neutrophil sequestration. Resuscitation with HTS ameliorated T/HS-induced gut and lung injury seen with RL resuscitation. These results, together with the direct correlation found between gut and lung injury, suggest that lung injury after T/HS may be mediated by gut injury.     

高渗氯化钠溶液复苏抑制失血性休克大鼠肺细胞凋亡的初步研究

目的 探讨高渗氯化钠溶液(HS)复苏对失血性休克大鼠肺细胞凋亡的影响及其意义. 方法 将23只SD大鼠制作成重度失血性休克模型,随机分为假手术组(Sham组,8只)、高渗氯化钠溶液复苏组(HS组,9只)和等渗盐水复苏组(NS组,6只),采用流式细胞仪FITC-AnnexinV/PI荧光染色法定量测定休克/复苏后各组大鼠肺组织细胞的凋亡情况,并加以比较和分析. 结果 在失血性休克/复苏后的早期阶段,HS组和NS组大鼠的肺组织细胞即有大量凋亡发生,其肺细胞凋亡率均明显高于Sham组,差异有统计学意义(P＜0.01).同时,NS组大鼠的肺细胞凋亡率则显著高于HS组,差异有统计学意义(P＜0.01). 结论 在重度失血性休克大鼠模型中,与等渗盐水复苏相比较,高渗氯化钠溶液复苏能显著抑制失血/复苏后肺细胞的凋亡,有助于减轻休克后急性肺损伤,这可能也是高渗氯化钠溶液复苏肺保护作用的重要机制之一.     

Serine proteases are involved in the pathogenesis of trauma-hemorrhagic shock-induced gut and lung injury

The objective of this work was to test the hypothesis that Intraluminal serine proteases are involved in trauma-hemorrhagic shock (T/HS)-induced intestinal and lung injury. Male Sprague-Dawley rats were administrated the serine protease inhibitor (6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfate, Nafamostat) either intraluminally into the gut or intravenously after a laparotomy (trauma) and then subjected to 90 min of hemorrhagic shock (T/HS) or sham shock (T/SS). Intestinal and lung injury was assessed at 3 h after resuscitation with Ringer's lactate solution. In a second set of experiments, mesenteric lymph was collected from the groups of rats subjected to T/HS or T/SS and its ability to activate normal neutrophils was tested. Lung permeability, pulmonary myeloperoxidase levels, and the bronchoalveolar lavage fluid protein to plasma protein ratio were increased after T/HS but were significantly decreased in the T/HS rats receiving intraluminal (P < 0.05), but not intravenous, nafamostat. Likewise, T/HS-induced intestinal villus injury was less in the nafamostat-treated shock rats (P < 0.05). Last, the ability of T/HS mesenteric lymph to increase PMN CD11b expression or prime neutrophils for an augmented respiratory burst was significantly reduced by the intraluminal administration of nafamostat. Because intraluminal nafamostat reduced T/HS-induced gut and lung injury as well as the neutrophil activating ability of intestinal T/HS lymph, the presence of serine proteases in the ischemic gut may play an important role in T/HS-induced gut and hence lung injury.     

Erythropoietin attenuates the tissue injury associated with hemorrhagic shock and myocardial ischemia

Here we investigate the effects of erythropoietin (EPO) on the tissue/organ injury caused by hemorrhagic shock (HS), endotoxic shock, and regional myocardial ischemia and reperfusion in anesthetized rats. Male Wistar rats were anesthetized with thiopental sodium (85 mg/kg i.p.) and subjected to hemorrhagic shock (HS; i.e., mean arterial blood pressure reduced to 45 mmHg for 90 min, followed by resuscitation with shed blood for 4 h), endotoxemia (for 6 h), or left anterior descending coronary artery occlusion (25 min) and reperfusion (2 h). HS and endotoxemia resulted in renal dysfunction and liver injury. Administration of EPO (300 IU/kg i.v., n = 10) before resuscitation abolished the renal dysfunction and liver injury in hemorrhagic, but not endotoxic, shock. HS also resulted in significant increases in the kidney of the activities of caspases 3, 8, and 9. This increase in caspase activity was not seen in HS rats treated with EPO. In cultured human proximal tubule cells, EPO concentration-dependently reduced the cell death and increase in caspase-3 activity caused by either ATP depletion (simulated ischemia) or hydrogen peroxide (oxidative stress). In the heart, administration of EPO (300 IU/kg i.v., n = 10) before reperfusion also caused a significant reduction in infarct size. In cultured rat cardiac myoblasts (H9C2 cells), EPO also reduced the increase in DNA fragmentation caused by either serum deprivation (simulated ischemia) or hydrogen peroxide (oxidative stress). We propose that the acute administration of EPO on reperfusion and/or resuscitation will reduce the tissue injury caused by ischemia-reperfusion of the heart (and other organs) and hemorrhagic shock.     

腹腔复苏对失血性休克大鼠肺损伤的保护作用

目的探讨腹腔复苏对失血性休克大鼠肺损伤的保护作用。方法健康雄性SD大鼠40只,随机分成假手术组(Ⅰ组)、失血性休克组(Ⅱ组)、静脉复苏组(Ⅲ组)、PD-2液腹腔复苏加静脉复苏组(Ⅳ组)。Ⅰ组依次行右颈总动脉、右股静脉、左股动脉插管及全身肝素化;Ⅱ组在Ⅰ组基础上制备失血性休克大鼠模型;Ⅲ组在Ⅱ组基础上于造模后经右股静脉补入放出的血量及2倍量林格氏液进行静脉复苏;Ⅳ组在Ⅲ组基础上同时腹腔内注入30 mL2.5%PD-2液。各组分别于T0、T1、T2进行PaO2/FiO2值血气分析;T2时测定肺组织W/D比值,TNF-α、IL-1β、IL-10的含量,观察肺组织病理学结果。结果与Ⅰ组比较,Ⅱ组PaO2/FiO2值明显降低,Ⅲ组于T0、T2时下降明显,Ⅳ组T0时下降明显;与Ⅰ组比较,Ⅱ组、Ⅲ组、Ⅳ组肺组织TNF-α、IL-1β、IL-10的含量和W/D比值均升高(P<0.01),与Ⅱ组比较,Ⅲ组和Ⅳ组降低,Ⅳ组肺组织IL-10含量升高(P<0.05或P<0.01)。与Ⅳ组比较Ⅲ组肺组织IL-10含量降低;肺组织病理学比较,Ⅱ组较Ⅰ组有明显损伤,Ⅳ组病理损伤最轻。结论 PD-2液腹腔复苏能够减轻失血性休克大鼠肺组织损害的程度,降低炎症反应,对肺损伤具有保护作用。     

不同液体复苏对未控制失血性休克大鼠肺损伤及肺水通道蛋白表达的影响

目的 观察不同液体复苏对未控制失血性休克大鼠肺损伤和肺水通道蛋白l(AQP1)和AQP5表达的影响.方法 SD大鼠被随机分为假手术组(C组)、无液体复苏组(NF组)、乳酸林格液组(LRS组)、高渗盐水组(HS组)和羟乙基淀粉组(HES)5组,每组12只.建立未控制失血性休克大鼠模型,模拟临床分为4期;动态观察各期的平均动脉压(MAP)变化.①失血性休克期(时间为60 min):在15 min内将MAP降至40 mm Hg(1mm Hg=0.133 kPa)并维持60 min;然后向气管内注射内毒素2 mg/kg,并用断尾法造成大鼠未控制失血性休克.②液体复苏期(时间为30 min):LRS、HS、HES组分别用3倍放血量的LRs、4 ml/kg的7.5%NaCl和1倍放血量的6%HES 130/0.4进行复苏,C组和NF组不处理.③综合复苏期(时间为1 h):在1 h内回输全部失血及1:1失血量的生理盐水.④复苏后观察期:输血、输液结束后继续观察3.5 h. 实验结束后测定肺组织湿/干重(W/D)比值;用免疫组化测定肺组织AQPl和AQP5表达,用苏木素一伊红(HE)染色,光镜下观察肺损伤程度.结果 在C组和HES组,肺血管内皮细胞AQP1和肺泡上皮AQP5均呈阳性表达;HS组仅AQP1呈阳性表达,AQP5则呈微弱表达;NF组和LRS组AQP1、AQP5均呈微弱表达.各组MAP、肺W/D比值和肺组织损伤程度比较显示,HS组和HES组显著优于NF组和LRS组(P<0.05或P<0.01).结论 遭受"二次打击"失血性休克大鼠并发肺损伤时,肺AQP1和AQP5表达下调;采用6%HES 130/0.4进行休克复苏可有效抑制AQP1和AQP5下调,并使肺损伤明显减轻;采用7.5%NaCI复苏仅能抑制AQPl表达下调和在一定程度上减轻肺水肿;而采用LRS复苏既不能保持AQP1、AQP5的表达,也不能有效防止肺损伤的发生.     

Resuscitation with hydroxyethyl starch solution prevents nuclear factor kappaB activation and oxidative stress after hemorrhagic shock and resuscitation in rats

Fluid resuscitation is vital for treating traumatic hemorrhagic shock (HS), but reperfusion is believed to have the adverse consequences of generating reactive oxygen species and inflammatory cytokines, both of which cause multiple organ dysfunctions. We investigated the effects of various resuscitation fluids on the changes of redox-sensitive molecules after HS and fluid resuscitation (HS/R). We induced HS by bleeding male Sprague-Dawley rats to a blood pressure of 30 to 40 mmHg for 60 minutes. Thirty minutes later, the rats were killed (HS group) or immediately resuscitated with shed blood (HS + BL group), L-isomer lactated Ringer's solution (HS + LR group), or hydroxyethyl starch (HS + HES group). After HS, we found a significant increase in nuclear factor kappaB DNA binding activity, which was effectively inhibited using HES solution or blood resuscitation. Moreover, resuscitation with blood or LR solution, but not HES solution, induced significant oxidative stress, manifested by a high ratio of oxidized glutathione to reduced glutathione in the lungs, liver, and spleen. HS alone, however, did not increase the ratio of the oxidized glutathione to reduced glutathione in all organs. Although the protein expression of anti-apoptotic Bcl-2 and pro-apoptotic Bax varied in different organs, we found that resuscitation using HES solution prevented the HS-induced reduction of the Bcl-2/Bax ratio in the heart. HES solution was an appropriate resuscitation fluid in reversing nuclear factor kappaB activation, maintaining the Bcl-2/Bax ratio, and preventing oxidative stress after acute HS.     

不同方式液体复苏对失血性休克大鼠外周血单个核细胞中核转录因子-κB活性的影响

目的观察失血性休克大鼠外周血单个核细胞（PBMC）中核转录因子-κB（NF—κB）活性的变化以及不同方式液体复苏对其的影响。方法将32只成功复制的失血性休克模型SD大鼠随机分为对照组、无液体复苏组、限制性液体复苏组和快速大量液体复苏组，每组8只；比较各组的救治疗效，并用酶联免疫吸附法（ELISA）检测各组PBMC中NF—κB活性的变化。结果限制性液体复苏组大鼠的存活时间较快速大量液体复苏组及无液体复苏组明显延长（P均〈0．05）；限制性液体复苏组大鼠72h存活率明显高于快速大量液体复苏组和无液体复苏组，但低于对照组（P均〈0．05）。除对照组外，其余各组创伤后60min和120min PBMC中NF—κB活性均较创伤前有明显升高，且120min较60min也明显升高（P均〈0．05）；限制性液体复苏组NF～κB活性明显低于快速大量液体复苏组和无液体复苏组（P均〈0．05）；死亡组创伤后60min和120min NF—κB活性明显高于存活组（P均〈0．05）。结论限制性液体复苏可显著降低失血性休克大鼠的72h死亡率；PBMC中NF—κB活性与预后密切相关，NF—κB活性高则提示预后不良，而限制性液体复苏时NF-κB活性明显降低，有助于改善预后。