一种新的复合型野战创伤早期抗休克液的实验研究

目的 依据失血性休克发生的理论基础和长期的临床实践经验,配制一种新的"复合型战地抗休克液",并利用大鼠失血性休克模型进行评价.方法 建立大鼠失血性休克模型.以乳酸林格液和羟乙基淀粉的2∶1混合液(LH)作为对照组休克复苏液;以添加地塞米松和呋塞米的LH作为加药组休克复苏液;复苏4 h后取血和器官标本检测生化、肝功能指标和肺组织含水量,并观察组织病理学变化.结果 两组液体均可使失血性休克大鼠平均动脉压恢复到基础值.与对照组比较,加药组血生化指标均无明显差异,但大鼠复苏后肺组织湿/干重比值明显降低(4.56±0.14比4.88±0.29,P＜0.05),丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平显著下降[ALT:(73.02±41. 89)U/L比(193.85±104.49)U/L;AST:(199.06±108.70)U/L比(395.25±137.08)U/L,均P＜0.05],利尿作用明显[开始排尿时间:(76±20)min比(153±14)min;尿量:(9.6±5.2)ml比(1.5±2.2)ml,P＜0.01和P＜0.05],腹水量减少[(1.3±0.6)ml比(5.0±3.0)ml,P＜0.05].肝、肺和小肠组织病理学观察也显示加药组损伤轻于对照组.结论 本研究中设计的"复合型战地抗休克液"配方简单,用于大鼠失血性休克复苏可快速恢复并稳定血流动力学,同时可起到减轻组织水肿及肝、肺、小肠损伤的作用.

Abstract：

Objective To propose a resuscitation fluid with a new formula for resuscitation of shock in battlefield on the basis of pathogenesis of hemorrhagic shock and clinical experiences, and to evaluate its safety and effectiveness in a rat hemorrhagic shock model. Methods After hemorrhagic shock was (referred to as LH)was used for resuscitation in animals of control group; LH supplemented with dexamethasone and furosemide(referred to as LHDF)was used as resuscitation fluid for experimental group.After 4 hours of infusion, blood and major organs were obtained for serum biochemical tests, lung water content measurement and histopathological observation. Results The mean arterial pressure of rats of both control and experimental groups recovered rapidly after resuscitation. There was no significant difference in the parameters of serum biochemistry between control group and experimental group. The wet/dry weight ratio of lung tissue in experimental group was significantly lower than control group(4. 56±0. 14 vs. 4. 88±0. 29, P＜0. 05). The blood alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels in experimental group were also significantly lower than in control group[ALT:(73. 02±41.89)U/L vs.(193. 85 ± 104. 49)U/L; AST:(199.06 ± 108. 7)U/L vs.(395.25 ± 137.08)U/L, both P＜0. 05).Diuretic effect was obviously observed in experimental group than control group[time of start urination:(76±20)minutes vs.(153±14)minutes; urine volume:(9. 6±5. 2)ml vs.(1.5±2. 2)ml, P＜0. 01 and P＜0. 05], and the amount of ascitic fluid in experimental group was significantly lower than in control group [(1.3 ± 0. 6)ml vs.(5.0 ± 3.0)ml, P＜ 0. 05). Histopathological observation of the liver, lung and intestine also showed less pathological changes in experimental group than in control group. Conclusion The designed battlefield anti-shock fluid in this study has been shown to be effective in fluid resuscitation for hemorrhage shock in rats, with reduced tissue edema and less injury to the liver, lung and intestine.     

The studies on the changes of nitric oxide synthase in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the secondary lung injuris北大核心CSCD

Objective: To study the pathological changes and expressions of NO and iNOS mRNA in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the lung injury. Methods: A total of 140 male SD rats were randomly (random number) ivided into the room temperature (25°C) environment traumatic hemorrhagic shock group (room temperature group) and the dry heat traumatic hemorrhagic shock groups (dry heat group, temperature 40°C, humidity 10%), respectively, and each groups was further randomly divided into 7 subgroups: the control subgroup, post shock subgroups at 0, 0.5, 1, 1.5, 2 and 3 h (re = 10 in each subgroup). The rats of control subgroup were not treated, and rats of dry heat group were placed in dry heat environment for 60 min, then anesthetized, fixed, and insertion of intravenous indwelling needles and catherization of right carotid artery, jugular vein and the right femoral artery were performed. After stabilization for 10 min, 2500 g iron wheel was used to be dropped from 30 m height and vertically hit the upper left femoral of SD rats in order to make comminuted fracture, wounds were quickly dressed after injury. Exsanguination from right femoral artery was kept until MAP maintained at (35 ± 5) mmHg, and resuscitation was carried out after continue monitoring for 60 min. After the establishment of traumatic hemorrhagic shock model in each environment, the rats were sacrificed at given intervals, and thoracotomy was performed to take broncho - alveolar lavage fluid (BALF) and lung tissue. Pathological changes of lung tissues were observed by using HE staining and NO concentration of lung tissue was detected by one-step method, and changes of the iNOS mRNA expressions were detected by using fluorescence quantitative PCR. Then t test, ANOVA and Pearson correlation analysis were used for the data analysis. Results: The pathological change in dry heat group at each interval was more severe, and pulmonary histopathological injury score was higher, and the protein exudation was more profuse compared with the room temperature group. NO concentration in lung tissue homogenate of dry heat group was higher than that of room temperature group (t = 2.472, P < 0.05), and the difference in NO level between different intervals within the dry heat group was statistically significant (F = 6.11, P < 0.01). The NO concentration in dry heat group reached its maximum at 2 h (3.35 ± 0.23) μmol/g and the peak value emerged sooner than that in room temperature group. The difference was statistically significant in overall expression of iNOS mRNA between two groups analyzed with t test (t = 3.619, P < 0.01), and there was statistically significant difference between intervals within the dry heat group (F = 12.34, P < 0.01). The values of iNOS mRNA in the dry heat group were higher than those in the room temperature group at the same given intervals, and the peak value appears at 1.5 h in dry heat group, and the room temperature group it began to increase at 2 h. The concentration of NO and the expression of iNOS mRNA were positively correlated with each other in two groups (r = 0.680, r = 0.376). The expression of iNOS mRNA and lung histopathological injury score was positively correlated in two groups (r = 0.846, r = 0.899). Conclusions When traumatic hemorrhagic shock occurred in the dry heat desert environment, the lung injury was more severe and appeared sooner than that in the room temperature environment. NO and iNOS played important roles in the secondary lung injury in the wake of traumatic hemorrhagic shock in rats under the dry heat environmengt of desert.     

脉搏轮廓法在感染性休克早期液体复苏中的运用

目的 探讨以脉搏轮廓动脉压波形分析法(PiCCO)指标指导感染性休克患者液体复苏及血管活性药应用及与预后的关系.方法 2006年12月至2008年6月感染性休克患者为常规治疗组,共纳入30例;2008年7月至2009年10月为PiCCO组,共纳入26例.排除标准为既往有慢性心肺及肝肾功能不全病史.常规组给予EDGT方案复苏.PiCCO组根据每搏变异量及全心舒张末期容积指数进行液体复苏,使全心舒张末期容积指数达到600～750 mL/m2,无房颤时同时考虑每搏变异量＜10%;并根据体循环外周阻力指数监测结果应用去甲肾上腺素调整在1300～1500 d·s·cm-5·m-2,根据全心射血分数及心脏指数应用多巴酚丁胺调节心功能,维持平均动脉压≥65 mmHg,同时监测血管外肺水指导液体选择和利尿剂应用.观察患者6 h后中心静脉氧饱和度和乳酸值、3 d的液体平衡状态、28 d生存率、28 d脱离呼吸机时间、住ICU时间和无脏器衰竭时间.结果 两组患者的年龄、性别、病因、APACHEⅡ及SOFA评分、血流动力学状态在液体复苏前差异无统计学意义.PiCCO组与常规组液体复苏6 h后中心静脉氧饱和度和乳酸值差异无统计学意义,28 d生存率和无脏器衰竭时间比较差异无统计学意义(P＞0.05).PiCCO组6 h和1 d血管活性药物多巴酚丁胺量[(145.4±24.5)mg vs.(104.2±46.3)mg;(330.2±30.3)mg vs.(202.4±40.3)mg]及去甲肾上腺素用量[(14.5±3.8)mg vs.(10.2±5.6)mg;(38.2±4.2)mg vs.(20.1±6.2)mg]较常规组明显要多(P＜0.05);但2d与3 d血管活性药量差异无统计学意义(P＞0.05).PiCCO组6 h及3 d液体复苏的量明显减少[(2121±578)mL vs.(2910±987)mL;(3845±435)mL vs.(4545±765)mL;(2467±510)mL vs.(2867±618)mL;(951±332)mL vs.(1472±533)mL],28 d内脱离呼吸机时间[(19.7±8.3)d vs.(15.1±9.1)d]明显增加,住ICU时间明显缩短[(7.5±3.5)d vs.(9.5±3.2)d](P＜0.05).结论 以PiCCO的指标指导感染性休克患者液体复苏及血管活性药应用,能更精确进行液体管理,减少盲目补液,减少机械通气时间,缩短住ICU时间.

Abstract：

Objective To explore the use of pulse indication contour cardiac output (PiCCO) as a guidance for fluid resuscitation and vasopressors employment in patients with septic shock in order to find out the efficacy of this resuscitation strategy in the respect of outcome of patients. Mtthod A total of 30 patients with septic shock were treated with the EDGT fluid treatment protocol as the conventional treatment group from December 2006 to June 2008; and another 26 patients were given fluid treatment under the guidance of PiCCO as PiCCO group from July 2008 to October 2009. Exclusion criteria included patients with history of heart and lung diseases, and liver and kidney dysfunction. The mean arterial blood pressure was maintained above or equal to 65mmHg in PiCCO group,and fluid resuscitation was concluded when global end-diastolic volume index reached 600～750 mL·-2 with the stroke volume variation in ＜ 10% and without auricular fibrillation. Nor-epinephrine was administrated to adiust the systemic peripheral vascular resistance index during 1300～1500 d·s·cm-5·m-2.Dobutamine was empoyed when global ejection fraction was compromised. The options of liquid and diuretics depend upon the presence d extra-vascular lung water. Central venous oxygen saturation and the level of lactate were observed 6hours after resuscitation. The liquid equilibrium for 3 days and the dosage of vasopressors were also recorded. The rate of survival, the time taken for weaning from mechanical ventilation, the days of ICU stay and rate of intact organ function within 28 days were compared between two groups. Results The demphics of patients of two goups were similar. There were no significant difference between PiCCO and the conventional group in values of central venous oxygen saturation and lactate 6 hours after admission to ICU (P ＞ 0.05). And 6 h and 1d after fluid resuscitation, the dosages of dobutamine and the dosages of nor-epinephrine used in PiCCO group were significantly higher than those in conventional group in which the dosages of dobutamine in two groups were [(145.4±24.5)mg vs. (104.2 ± 46.3) mg and (330.2 ± 30.3) mg vs. (202.4 ± 40.3) mg], respectively, and the dosages of nor-epinephrine [(14.5±3.8) mgvs. (10.2±5.6) mgand (38.2±4.2) rng vs.(20.1±6.2) mg], respectively. However, the dosages of vasopressors were similar between two groups 2 d and 3 d later. The amounts of liquid administered to get equilibrium in 6 h and the 3 d in PiCCO group were significantly less than those in conventional group [(2121±578) mL vs. (2910±987) mL and (3845±435) mL vs. (4545 ± 765) mL and (2467±510) mLvs. (2867±618) mL and (951±332) mLvs. (1472±533) mL], respectively. The days required to get free from mechanical ventilation within 28d were significantly longer in PiCGO group[(19.7 ± 8.3) d vs.(15.1±9.1) d], but the days of ICU stay were significantly shorter in PiCCO group [(7.5 ± 3.5) d vs. (9.5±3.2) d] (P＜0.05). The rates of survival and days free from organ failure within 28 days of two groups were similar(P＞0.05). Conclusions When the early fluid management guided with PiCCO in septic shock patients,the fluid management can be implement more safely and precisely. It can shorten the days of ICU stay and days of mechanical ventilation support with avoidance of fluid resuscitation.     

C5aR和P38-MAPK在脓毒性休克大鼠心肌损伤中的作用

目的 研究补体C5a受体与P38-MAPK在脓毒性休克情况下诱导心肌损伤的关系.方法 采用盲肠结扎剪切法构建早期脓毒性休克大鼠动物模型,实验地点在武汉大学人民医院麻醉科实验室.30只Sprague-Dawley大鼠随机(随机数字法)分为正常对照组6只和模型组24只(12 h组12只,24 h组12只).12 h组于术后12 h时点检测血清乳酸脱氢酶(LDH)和肌酸激酶(CK)水平,12 h后麻醉处死大鼠,迅速开胸取出心脏组织,行HE染色、应用免疫组织化学方法检测C5a受体和P38-MAPK的表达情况.24 h组亦于术后24 h时点检测血清LDH和CK值,24 h后处死大鼠取心肌组织行HE染色、检测C5a受体和P38-MAPK的表达情况.结果 模型组(12 h组和24 h组)与正常对照组相比,LDH值和CK值均明显升高(P＜0.05).24 h组LDH值和CK值与12 h时间点相比,差异具有统计学意义[(2 568.9±280)vs.(2 201.2±149),(5 029.7±458)vs.(2 629.4±140),P＜0.05].C5aR和P38-MAPK灰度值分析显示,模型组与正常对照组相比,均有显著性增高(P＜0.05),24 h组较12 h组相比,差异具有统计学意义[(702.77±122)vs.(388.36±113),(646.40±181)vs.(307.32±61),P＜0.05].相关分析显示C5aR和P38-MAPK存在显著正相关关系,(P＜0.05),P38-MAPK与LDH和CK均存在显著正相关关系(P＜0.05).结论 C5a受体和P38-MAPK在诱导脓毒性休克心肌损伤中有着强大的协同效应.

Abstract：

Objective To investigate effects of complement C5a receptor and P38-MAPK on myocardial injury brought about by septic shock in rats. Method The early septic shock models were established by the method of cecal ligature and incision (CLI). A total of 30 Sprague-Dawley rats were randomly( random number) divided into normal control group ( n = 6 ) and model group ( n = 24 ) and the model group was further 12 hours later divided into 12 h subgroup (n = 12) and 24 h subgroup (n = 12). The arterial blood samples were collected 12 hours later for detecting the levels of lactate dehydrogenase (LDH) and creatine kinase (CK), and then the rats were sacrificed and the myocardial tissues were taken to assay the expressions of C5a receptor and P38-MAPK by using immunohistochemistry after HE staining. And the above procedure as did in 12 h subgroup was done 24 hours later. Results Compared with the control group, the levels of LDH and CK in rats of Model group were significantly higher (P ＜ 0. 05). There were significant differences in LDH and CK between 24 h subgroup and 12 h subgroup [(2 568.9 ± 280) vs. (2 201.2 ± 149)] and [(5 029.7±458) vs. (2 629.4±140)] ,P＜0. 05, P＜0.05. The analysis of C5aR and P38-MAPK gray values showed that there were significant differences between the model group and normal control group [(702.77 ±122) vs. (388.36±113)], P＜0. 05 and [(646.40±181) vs. (307.32 ±61)] ,P＜0.05,and those differences also found between the 24 h subgroup and 12 h subgroup. There was a significant positive correlation between C5aR and P38-MAPK (P＜0.05 ), and also the P38-MAPK had significant positive relationships with LDH(P＜0.05) and CK (P＜0.05). Conclusions The C5aR strongly potentiates the P38-MAPK to induce myocardial injury by septic shock.     

膜攻击复合物C5b-9对创伤失血性休克大鼠肝损害的影响研究

目的 观察大鼠创伤失血性休克时肝脏是否受膜攻击复合物攻击,以及膜攻击复合物是否对肝脏细胞凋亡产生影响.方法 雄性Wistar大鼠50只,按随机数字表法均分为正常组及模型1、3、6、24 h组5组.采用骨折后经颈动脉放血至血压40 mm Hg(1 mm Hg=0.133 kPa)制备创伤失血性休克模型.取血浆,采用酶联免疫吸附法(ELISA)检测膜攻击复合物C5b-9浓度,采用速率法测定丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)浓度;取肝脏组织,采用免疫组化法检测C5b-9阳性表达,用原位末端缺刻标记法(TUNEL)检测肝细胞凋亡,苏木素-伊红(HE)染色,光镜下观察病理改变.结果 正常组血中能检测到少量的C5b-9;模型1、3、6 h组血中C5b-9浓度(ng/L)较正常组显著升高(272.91±9.56、192.01±9.04、156.78±8.37比25.98±5.87,均P＜0.05).模型3 h组血ALT(U/L)、模型1 h组血AST(U/L)即显著上升(92.90±8.83、264.83±31.14),24 h达高峰(184.30±12.98、647.36±60.02),与正常组(38.75±5.40、66.69±19.95)比较差异均有统计学意义(均P＜0.05).正常组及模型1 h、6 h组肝脏未发现C5b-9阳性表达;模型3 h组门管区大量肝脏实质细胞存在C5b-9(个)沉积(22.60±1.06),模型24 h组C5b-9沉积明显减少(2.20±0.60,P＜0.05).正常组未检测到凋亡细胞;模型1、6、24 h组发现散在凋亡细胞(个:1.20±0.25、5.60±0.37、1.60±0.26),模型3 h组中央静脉周围凋亡细胞明显增加,出现凋亡高峰(20.60±0.47),与其余各组比较差异均有统计学意义(均P＜0.05).模型组可见肝细胞水肿变性、肝细胞膜完整性破坏,细胞溶解,以24 h病理损害最重.结论 创伤失血性休克时大鼠肝脏受到膜攻击复合物C5b-9的攻击,并且肝脏C5b-9的表达高峰与凋亡高峰在同一时间点出现,但并不是同一部位;模型3 h后血中低水平的C5b-9提示预后极差.

Abstract：

Objective To observe whether the membrane attack complex C5b-9 would accumulate in the rats' liver after receiving the assault of traumatic hemorrhagic shock, and whether the membrane attack complex deals an impact on liver apoptosis. Methods Fifty male healthy Wistar rats were randomly divided into five groups: normal group, 1, 3, 6, 24-hour model groups. The model of traumatic hemorrhagic shock was reproduced by withdrawal of blood from carotid artery after a bone fracture till the blood pressure lowered to 40 mm Hg(1 mm Hg= 0. 133 kPa). Plasma membrane attack complex C5b-9 concentration was assayed using enzyme linked immunoadsorbent assay. Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in blood was determined by Rate method. Immunohistochemistry was used to detect C5b-9 deposition in the liver. Apoptosis of liver cells was then detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay. The pathological changes in paraffin sections stained with hematoxylin-eosin(HE)were observed under light microscope. Results A small amount of C5b-9 in plasma was found in normal group, and the values(ng/L)of 1, 3, 6-hour models were significantly higher than those of the normal group(272. 91±9.56, 192. 01±9. 04, 156. 78±8. 37 vs. 25. 98±5.87, all P＜0. 05). ALT(U/L)in 3-hour model group and AST(U/L)in 1-hour model group were increased significantly(92.90 ± 8. 83, 264.83 ± 31.4), peaked at 24 hours(184.30 ± 12. 98, 647.36 ± 60. 02), and there was significant difference compared with normal group(38. 75±5.40, 66. 69± 19.95, all P＜0. 05). In the normal group and the 1-hour and 6-hour model groups, no C5b-9 was found in liver, but in the 3-hour model group a large number of liver parenchymal cells in the portal area were found to contain C5b-9 (22. 60± 1.06), however the number decreased significantly in the 24-hour model(2. 20±0. 60, P＜0. 05).In normal group there was no apoptotic cell, and in 1, 6, 24-hour model groups there were scattered apoptotic cells(1. 20±0. 25, 5. 60±0. 37, 1. 60±0. 26). In the 3-hour model group apoptosis of hepatic cells around the central vein was increased to the peak(20. 60 ± 0. 47), and there was significant difference compared with other groups(all P＜0. 05). In the model groups the liver cells became edematous, and the integrity of the membrane was lost, and some cells were even lysed. The pathological damage is most serious in 24-hour model group. Conclusion The membrane attack complex C5b-9 insulted the rats' liver after a traumatic hemorrhagic shock, and apoptosis of hepatic cells and the content of C5b-9 peaked in 3-hour model, though they do not occur in the same site. A low level of C5b-9 in blood 3 hours after shock predict a poor prognosis.     

前列腺素E1预先给药对出血性休克复苏后大鼠肾组织Caspase-3表达的影响

目的 探讨PGE1预先给药对急性出血性休克复苏后大鼠肾组织活化Caspase-3以及TNF-α mRNA,iNOS mRNA表达的影响.方法 32只雄性SD健康大鼠随机分成4组(n=8),A:正常对照组,不进行任何处理;B:手术对照组,0.5 mL生理盐水尾静脉注射,1 h后切开左侧腹股沟皮肤暴露股动静脉;C:出血性休克复苏组,大鼠硫喷妥纳麻醉后,经股静脉穿刺置管放血,平均动脉血压降至25～35 mmHg,维持1 h,自体温血复苏30 min,维持平均动脉血压80～100 mmHg,制作失血性休克复苏模型;D:出血性休克复苏+lipo-PGEl预先给药组,lipo-PGEl按10 μg/kg溶于0.5 mL生理盐水尾静脉汴射,1 h后制作出血性休克复苏模型.各组分别于复苏后6 h时间点用Northern blotting法检测肾组织TNF-α mRNA和iNOS mRNA的表达;免疫组化检测肾脏组织活化Caspase-3的表达.数据采用单因数方差分析(SNK-q检验),以P<0.05为差异具有统计学意义.结果 出血性休克复苏模型组肾组织出现Caspase-3阳性表达细胞;TNF-α mRNA和iNOS mRNA的表达也明显高于正常对照组[(0.651±0.028)vs.(0.030±0.003),(0.524±0.022)vs.(0.026±0.003),P<0.05]和手术对照组[(0.651±0.028)vs.(0.029±0.002),(0.524±0.022)vs.(0.025±0.003),P<0.05];预先给药组肾组织Caspase-3阳性细胞数量显著减少;TNF-αmRNA和iNOS mRNA的表达明显低于模型组[(0.250±0.019)vs.(0.651±0.028),(0.203±0.020)vs.(0.524±0.022),P<0.05],正常对照组和手术对照组间肾组织Caspase-3的表达以及TNF-α mRNA和iNOS mRNA的表达尢显著差异[(0.029±0.002)vs.(0.030±0.003),(0.029±0.002)vs.(0.030±0.003),P>0.05].结论 lipo-PGEl预先给药可以下调出血性休克复苏后大鼠肾组织Caspase-3的表达水平,其机制可能与在基因转录水平抑制TNF-α和I-NOS的表达有关.     

C5aR和P38-MAPK在脓毒性休克大鼠心肌损伤中的作用

Objective To investigate effects of complement C5a receptor and P38-MAPK on myocardial injury brought about by septic shock in rats. Method The early septic shock models were established by the method of cecal ligature and incision (CLI). A total of 30 Sprague-Dawley rats were randomly( random number) divided into normal control group ( n = 6 ) and model group ( n = 24 ) and the model group was further 12 hours later divided into 12 h subgroup (n = 12) and 24 h subgroup (n = 12). The arterial blood samples were collected 12 hours later for detecting the levels of lactate dehydrogenase (LDH) and creatine kinase (CK), and then the rats were sacrificed and the myocardial tissues were taken to assay the expressions of C5a receptor and P38-MAPK by using immunohistochemistry after HE staining. And the above procedure as did in 12 h subgroup was done 24 hours later. Results Compared with the control group, the levels of LDH and CK in rats of Model group were significantly higher (P ＜ 0. 05). There were significant differences in LDH and CK between 24 h subgroup and 12 h subgroup [(2 568.9 ± 280) vs. (2 201.2 ± 149)] and [(5 029.7±458) vs. (2 629.4±140)] ,P＜0. 05, P＜0.05. The analysis of C5aR and P38-MAPK gray values showed that there were significant differences between the model group and normal control group [(702.77 ±122) vs. (388.36±113)], P＜0. 05 and [(646.40±181) vs. (307.32 ±61)] ,P＜0.05,and those differences also found between the 24 h subgroup and 12 h subgroup. There was a significant positive correlation between C5aR and P38-MAPK (P＜0.05 ), and also the P38-MAPK had significant positive relationships with LDH(P＜0.05) and CK (P＜0.05). Conclusions The C5aR strongly potentiates the P38-MAPK to induce myocardial injury by septic shock.     

多烯磷脂酰胆碱对脓毒症大鼠肝脏的保护作用

目的 观察多烯磷脂酰胆碱(PPC)对脓毒症大鼠肝脏的保护作用.方法 将50只健康雄性SD 大鼠按随机数字表法分为正常对照组(10只)、模型组(15只)、PPC对照组(10只)和PPC保护组(15只).通过腹腔注射单剂大肠杆菌脂多糖(LPS)6 mg/kg建立脓毒症大鼠模型;正常对照组和PPC对照组给予等量生理盐水.PPC对照组和PPC保护组在注射LPS前24 h和6 h分别通过大鼠尾静脉给予等倍稀释的PPC 10 ml/kg(232.5 mg/kg);正常对照组和模型组给予等量的5%葡萄糖溶液.观察注射LPS后24 h内各组大鼠的行为改变和死亡率;处死动物后取静脉血检测丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST);留取肝脏测定大鼠全肝重/体重比值(L/Wb)和肝湿重/干重比值(W/D),并行组织病理学检查,用免疫组化二步法检测肝组织中细胞间黏附分子-1(ICAM-1)的表达.结果 正常对照组和PPC对照组大鼠精神状态正常;模型组大鼠精神萎靡,饮水、活动少;PPC保护组精神状态较好,饮水、活动较好.PPC保护组大鼠死亡率明显低于模型组[6.7%(1/15)比46.7 %(7/15),P<0.05].PPC保护组血浆ALT和AST水平、L/Wb比值、W/D 比值及肝组织ICAM-1阳性率均低于模型组[ALT(U/L):157.71±32.63比225.63±43.47;AST(U/L):53.21±13.85比85.25±18.91;L/Wb比值:(4.09±0.28)%比(4.50±0.25)%;W/D比值:3.52±0.27比3.84±0.18;ICAM-1阳性率:35.7%(5/14)比87.5%(7/8),P<0.05或P<0.01].正常对照组与PPC对照组上述指标未见统计学差异.模型组大鼠肝脏炎症反应强烈,炎性细胞聚集,肝实质细胞水肿渗出明显;正常对照组和PPC对照组肝脏形态正常;而PPC保护组介于两者之间.结论 大鼠预防性注射PPC能阻止内毒素引起的进行性炎症紊乱,减轻肝脏水肿和炎症反应,减少肝脏ICAM-1的表达,保护肝功能,并降低死亡率.

Abstract：

Objective To investigate the protective effect of polyenylphosphatidyl choline(PPC)on liver in rat with sepsis.Methods A total of 50 healthy male SD rats were randomly divided into four groups according to random number table:normal control group(NS group,n=10),sepsis model group(LPS group,n=15),PPC control group(PPC group,n=10)and PPC protection group(P+L group,n=15).Asingle dose of lipopolysaccharide(LPS)6 mg/kg was injected to the peritoneal cavity to reproduce the sepsis model,while in NS group and PPC group,same volume of normal saline was used.PPC 10 ml/kg (232.5 mg/kg)was injected 24 hours and 6 hours before LPS via the tail vein in PPC group and P+L group,while in NS group and LPS group 5%glucose solution in the same volume was given.Behavioral changes and mortality rate of rats were recorded,and all survived rats were sacrificed 24 hours after LPS injection.Venous blood was taken for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)determination.Ratios of liver weight/body weight(L/Wb)and wet wieght/dry weight of liver(W/D)were calculated.Histopathology changes in liver tissue were observed with hematoxylin eosin(HE)stain,and intercellular adhesion molecule-1(ICAM-1)expression with instant two-step immunohistochemistry.Results Twenty-four hours after LPS injection(LPS group),the rats became lethargic,with less activity and drinking,while rats in P+L group,showed much better mental status,water intake and activity than LPS group.The mortality rate of P+L group was significantly lower than that of LPS group[6.7%(1/15)vs.46.7%(7/15),P<0.05].Compared with LPS group,rats in P+L group had lower levels of plasma ALT and AST,L/Wb,W/D,and liver ICAM-1 positive expression ratio[ALT(U/L):157.71±32.63 vs.225.63±43.47; AST(U/L): 53.21±13.85 vs.85.25±18.91;L/Wb:(4.09±0.28)%vs.(4.50±0.25)%,W/D:3.52±0.27 vs.3.84±0.18,ICAM-1 positive expression ratio:35.7%(5/14)vs.87.5(7/8),P<0.05 or P<0.01].All parameters were normal in NS group and PPC group,and no statistical significance was observed between them(all P>0.05).LPS-injection led to severe inflammatory reaction in hepatic tissue,including obvious edema of hepatic parenchymal cells,exudation and aggregation of inflammatory cells,while in P+L group,these morphological changes were milder than that in LPS group but more obvious than that of NS group.Conclusion Pre-treatment of rats with PPC alleviates progressive inflammatory disturbances of hepafic tissue caused by endotoxin injection,and it attenuates liver edema and ICAM-1 expression,protects liver function and decreases mortality rate in rats with sepsis.     

Effect of ulinastatin on expressions of serum cardiac troponin Ⅰ, myocardial tumor necrosis factor-α and endothelin-1 in septic rats

Objective To investigate the myocardial protective effects of different dosage of ulinastatin (UTI) and the possible mechanism in septic rats.Methods Forty male Sprague-Dawley (SD) rats were randomly divided into five groups: control group, sham group, model group, UTI in low dose or high dose group.Cecal ligation and puncture (CLP) was adopted to reproduce animal model of sepsis.Left ventricular myocardium was harvested and blood samples were collected at 24 hours after successful establishment of animal model.Serum cardiac troponin Ⅰ (cTnI), the contents of myocardial tumor necrosis factor-α (TNF-α)and endothelin-1 (ET-1) were measured, and myocardial pathological changes were observed.Results In the model group, the level of serum cTnI, and the expressions of myocardial TNF-α and ET-1 were much higher than those in control group [cTnI (μg/L): 7.58 ± 0.53 vs.1.05 ± 0.21, TNF-α (pg/g): 945.6 ±72.0 vs.238.2±35.2, ET-1 (pg/g): 776.8±123.9 vs.170.1±28.3, all P＜0.01].There were no differences in the levels of serum cTnI, myocardial TNF-α and ET-1 between low dose UTI group and the model group [cTnI (μg/L): 7.21± 0.51 vs.7.58 ± 0.53, TNF-α(pg/g):910.5 ± 96.6 vs.945.6 ± 72.0,ET-1 (pg/g): 714.0±66.7 vs.776.8±123.9, all P＞0.05].However, serum cTnI, myocardial TNF-α and ET-1 were lower significantly in high dose UTI group than in model group [cTnI (μg/L): 4.30±0.84 vs.7.58±0.53, TNF-α(pg/g): 430.5±75.6 vs.945.6±72.0, ET-1 (pg/g): 377.1±39.0 vs.776.8±123.9, all P＜0.01].Conclusion High dose (but not low dose) UTI may protect myocardium from the damage resulted from sepsis in a rat model, probably by lowering expressions of TNF-α and ET-1.     

瑞舒伐他汀对兔心肌缺血-再灌注损伤内皮功能的影响

目的 研究兔心肌缺血-再灌注损伤中内皮功能改变及瑞舒伐他汀对其影响.方法 将16只新西兰大白兔随机分为两组:心肌缺血-再灌注损伤组(对照组),瑞舒伐他汀干预组(药物组).建立心肌缺血-再灌注模型,心电图显示ST段明显弓背向上抬高(≥0.2 mV)表示结扎左前降支成功;40 min后剪断结扎线,心电图ST段回落1/2以上,标志再灌注成功.分别在缺血前,缺血40 min,再灌注60 min和再灌注180 min四个时相点留取血标本,检测一氧化氮(nitric oxide,NO)、内皮素-1(endothe-lin-1,ET-1)含量.运用SPSS 11.5软件,采用重复测量资料的方差分析,以P<0.05为差异具有统计学意义.结果 对照组与药物组血清NO含量随缺血及再灌注时间的延长而降低,血浆ET-1含量随缺血及再灌注时间的延长而增高;缺血前对照组与药物组血清N0含量[(109.875±32.255)μmol/Lvs.(114.500±37.405)μmol/L,P>0.05]及血浆ET-1含量[(221.111±28.125)pg/mL vs.(204.594±31.790)pg/mL,P>0.05]差异无统计学意义;缺血40 min、再灌注60 min、再灌注180 min时,药物组血清NO含量[分别为(63.125±18.962)、(43.500±16.518)、(29.625±14.162)μmol/L vs.(82.000±13.825)、(63.375±17.541)、(50.250±18.987)μmol/L,P<0.05]均显著高于对照组,药物组血浆ET-1含量[分别为(331.785±35.341)、(375.914±45.204)、(459.829±70.110)pg/mL vs.(282.541±38.928)、(315.152±55.263)、(377.795±60.427)pg/mL,P<0.05]均显著低于对照组.结论 瑞舒伐他汀通过升高血清中NO,降低血浆ET-1,从而改善兔心肌缺血-再灌注损伤时内皮功能.     

肠系膜淋巴管结扎对失血性休克大鼠的器官保护作用

目的 观察肠系膜淋巴管结扎对失血性休克大鼠肠、肝、肺组织细胞因子表达以及组织病理学的影响.方法 24只SD大鼠被随机均分成对照组、失血性休克组、失血性休克+肠系膜淋巴管结扎组.采用逆转录一聚合酶链反应(RT-PCR)检测各组大鼠肠、肝、肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的mRNA表达;苏木素一伊红(HE)染色观察各组织的病理学改变.结果 失血性休克大鼠肠、肝、肺组织TNF-α mRNA和IL-6 mRNA表达均较对照组明显升高(TNF-α mRNA:肠0.54±0.07比0.37±0.05,肝1.014-0.06比0.56±0.07,肺0.94±0.07比0.62±0.06;IL-6 mRNA:肠0.89±0.12比0.50±0.09,肝1.07±0.10比0.57±0.12,肺1.09±0.09比0.67±0.06,P均<0.01);肠系膜淋巴管结扎可明显降低肠、肝、肺组织TNF-αmRNA和IL-6 mRNA表达(TNF-α mRNA:肠0.47±0.05比0.54±0.07,肝0.81±0.07比1.01±0.06,肺0.80±0.05比0.94±0.07;IL-6 mRNA:肠0.66±0.07比0.89±0.12,肝0.83±0.13比1.07±0.10,肺0.73±0.11比1.09±0.09,P<0.05或P<0.01).组织病理学观察显示,肠系膜淋巴管结扎可明显减轻失血性休克引起的肠黏膜绒毛坏死、脱落;减轻肝细胞变性、坏死;减轻肺水肿和炎性细胞浸润.结论 肠系膜淋巴管结扎可降低失血性休克大鼠肠、肝、肺组织细胞因子TNF-α、IL-6的表达及病理损伤程度,对器官功能起保护作用.     

阿魏酸钠对L-精氨酸诱导的重症急性胰腺炎大鼠的作用

目的 研究阿魏酸钠(soaium ferulate,SF)对L-精氨酸诱导的大鼠重症急性胰腺炎(severeacute pancreatitis,SAP)的治疗作用及其机制.方法 将60只成年SD大鼠随机分为对照组、SAP组、SF组,每组20只.采用腹腔内分两次注射大剂量L-精氨酸(2.5g/kg×2,间隔1 h)的方法制备SAP大鼠模型,SAP组和SF组大鼠分2次间隔1 h腹腔内注射20%L-精氨酸溶液(2.5 g/kg×2);对照组同法予等量生理盐水.5 min后,SF组大鼠再尾静脉注射SF注射液100 mg/kg,q d×3 d,其余两组同法予等量生理盐水.72 h后比较三组大鼠腹水性状、胰腺组织病理变化和血清淀粉酶(AMY)、内皮素-1(ET-1),TNF-α,IL-6水平和胰腺组织丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)含量及预后.计量资料组间比较用单因素方差分析,计数资料用Fisher确切概率法检验,以P<0.05为差异有统计学意义.结果 72 h后与对照组比较,SAP组出现典型的SAP病理形态改变,且大鼠血清AMY,ET-1,TNF-α,IL-6及胰腺组织NDA均显著升高[(9715.5±301.3)IU/L vs.(729.2±134.2)IU/L;(25.32±3.67)ng/L vs.(14.32±2.69)ng/L;(102.95±11.24)ng/L.vs.(38.62±3.87)ng/L;(538.63±9.53)ng/L vs.(186.35±1.19)ng/L;(34.8±3.9)mol/kg vs.(8.1±2.1)mol/kg,均P<0.01],胰腺组织抗氧化物质GSH,SOD明显降低[(7.1±0.6)mg/kg、vs.(16.9±1.9)mg/kg;(6423±1978)kU/kg vs.(29 905±z945)kU/kg,均P<0.01];与SAP组比较,SF组胰腺组织病理损害减轻且组织病理评分较低(P<0.05),血清AMY,ET-1,TNF-α,IL-6及胰腺组织NDA明显降低[(8104.6±149.9)IU/L vs.(9715.5±301.3)IU/L;(20.26±5.86)ng/L vs.(25.32±3.67)ng/L;(84.19±15.14)hg/L vs.(102.95±11.24)ng/L;(458±5.37)mol/kg vs.(538.63±9.53)ng/L;(28.3±2.5)mot/kg vs.(34.8±3.9)mol/kg,均P<0.05],胰腺组织GSH、SOD升高[(8.5±1.4)mg/kg vs.(7.1±0.6)mg/kg;(10316±2810)kU/kg、rs.(6423±1978)ldJ/kg,均P<0.05];SF组72 h死亡率低于SAP组,但差异无统计学意义(P=0.25).结论 SF能清除氧自由基、提高胰腺组织抗氧化物质SOD和GSH含量,降低血清促炎细胞因子及ET-1水平,减轻胰腺组织的病理损害,并且有可能降低死亡率,在SAP治疗中具有独特的优势.     

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.     

淋浆对内毒素休克大鼠肾肝血流灌注量变化的干预作用

目的 观察正常淋浆对内毒素休克(ES)大鼠肾、肝微区血流灌注量变化的干预作用. 方法 将30只Wistar大鼠按随机数字表法均分为对照组、模型组、淋浆组.采用静脉注射脂多糖(LPS,15 mg/kg)复制ES模型;对照组给予等量生理盐水.制模15 min后,淋浆组自颈静脉输入仅占全血量1/15 的淋浆;模型组和对照组以生理盐水代替.连续观察注射LPS后6 h内的颈动脉血压,用激光多普勒血流灌注仪连续记录肾、肝微区血流灌注量;并观察大鼠存活时间. 结果 注射LPS后,模型组及淋浆组血压迅速下降,回升波后模型组呈缓慢进行性下降,而淋浆组在休克早期及后期血压虽低于对照组,但显著高于模型组.模型组和淋浆组注射LPS后肾、肝微区血流灌注量(PU)显著低于对照组(肾:559.90±111.87、 577.50±91.49比672.60±50.81;肝:160.50±54.17、 163.56±34.04比232.30±34.23,均P<0.05).两组肾微区血流灌注量于30 min时回升并显著高于对照组,且淋浆组高水平维持至75 min,其后维持在对照组水平;而模型组120 min时显著降低,且于210 min起显著低于对照组.淋浆组肾微区血流灌注量(PU)于LPS后270～360 min显著高于模型组,以330 min后增高最为明显(330 min:615.44±98.71比364.40±146.76,360 min:584.56±104.72比307.11±143.11,均P<0.05).两组肝微区血流灌注量于30 min时恢复正常,模型组于LPS后270 min起呈进行性下降;淋浆组300 min后肝微区血流灌注量(PU)显著高于模型组,以330 min后增高最为明显(330 min:224.67±20.65比172.90±46.74,360 min:218.56±20.40比158.52±44.71,均P<0.05).模型组肾微区血流灌注量出现持续性低灌注的时间(min)显著早于肝脏(123.00±68.85比282.00±96.90,P<0.01).淋浆组动物存活时间(h)明显长于模型组(11.80±2.67比7.21±1.33,P<0.01). 结论 正常淋浆对ES时肾、肝组织低灌注具有良好的干预作用,并可改善低血压,延长存活时间.     

高渗盐溶液经骨髓腔通路复苏失血性休克犬的骨髓细胞学及病理学改变

目的 观察不同复苏液经骨髓腔通路复苏失血性休克后骨髓细胞学及病理学改变.方法 按随机数字表法将18只犬均分为假手术组、生理盐水组、高渗盐溶液组[即7.5%氯化钠-6%羟乙基淀粉40溶液(HSH)组].生理盐水和HSH组动物建立犬失血性休克模型,经胫骨骨髓腔快速(5 min内)按4 ml/kg分别推注生理盐水或HSH进行复苏;假手术组动物不进行放血和输液处理.于复苏前后取外周静脉血和骨髓液计数CD34+阳性率,骨髓涂片染色进行细胞分类及病理观察.结果 复苏后48 h,HSH组静脉血及骨髓液CD34+阳性率均高于生理盐水组(静脉血:(6.915±1.178)%比(4.848±0.527)%,骨髓液:(7.900±0.648)%比(6.875±0.403)%,均P<0.053.复苏后1周,与假手术组相比,生理盐水组红细胞系、粒细胞系细胞比例均显著升高(红细胞系:0.289±0.016比0.253±0.014,粒细胞系:0.615±0.019比0.560±0.013,均P<0.05),淋巴细胞系细胞比例显著下降(O.049±0.007比0.132±0.015,P<0.05);HSH组红细胞系(0.273±0.012)和淋巴细胞系比例(0.162±0.014)均显著升高(均P<0.05),而粒细胞系比例(0.517±0.038)显著下降(P<0.05).生理盐水组和HSH组4周后骨髓各造血细胞系恢复至正常;骨髓病理观察表明输液后骨髓改变轻微,未见坏死细胞.结论 经骨髓腔通路输注小剂量HSH复苏失血性休克犬后,骨髓改变轻微,是一种安全、有效的复苏方案.     

肠淋巴途径在大鼠休克致肝脏炎症反应中的作用

目的 观察结扎肠系膜淋巴管对重症失血性休克不同时期大鼠肝脏炎症介质、自由基的变化,探讨阻断肠淋巴途径对休克大鼠肝脏炎症反应的影响.方法 78只雄性Wistar大鼠被随机分为假手术组(n=6)、休克组(n=42)和结扎组(n=30).休克组与结扎组复制重症失血性休克模型,结扎组于休克复苏后行肠系膜淋巴管结扎术.于休克90 min、输液复苏后0、1、3、6、12和24 h各处死6只大鼠,制备肝组织匀浆,检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、一氧化氮(NO)、一氧化氮合酶(NOS)、丙二醛(MDA)、超氧化物歧化酶(SOD)以及髓过氧化物酶(MPO)水平;采用逆转录-聚合酶链反应(RT-PCR)测定肝组织诱生型一氧化氮合酶(iNOS)mRNA表达.结果 休克组大鼠输液复苏后不同时间点肝组织TNF-α、IL-6、NO、NOS、MDA、MPO以及iNOS mRNA均有不同程度的升高,6～12 h持续在较高水平,均显著高于假手术组,肝组织SOD活性显著低于假手术组(P＜0.05或P＜0.01);结扎组输液复苏后3、6、12和24 h肝组织TNF-α、IL-6、NO、NOS、MDA、MPO以及iNOS mRNA均显著低于休克组相应时间点.SOD活性高于休克组相应时间点(P＜0.05或P＜0.01).结论 肠系膜淋巴管结扎可减少肝脏中性粒细胞扣押,降低TNF-α、IL-6释放,抑制iNOS mRNA表达及NO生成,减少自由基损伤与SOD消耗,从而减轻肝脏的炎症反应.     

休克淋巴液对大鼠肺微血管内皮细胞炎症介质表达的影响

目的 观察休克淋巴液对大鼠肺微血管内皮细胞(PMVECs)自由基及一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达的影响,进一步探讨休克淋巴液损伤PMVECs的机制.方法 原代培养大鼠PMVECs至第3代进行研究.无菌条件下复制大鼠重症失血性休克模型(动脉压40 mm Hg维持90 min,1 mm Hg=0.133 kPa).引流正常大鼠和休克大鼠肠系膜淋巴液及门静脉血,与PMVECs孵育6h,同时以胎牛血清(FBS)和无血清的DMEM培养液作为对照.逆转录-聚合酶链反应(RT-PCR)检测诱生型一氧化氮合酶(iNOS)、TNF-α和IL-6的mRNA表达;检测培养上清液中丙二醛(MDA)、NO、TNF-α和IL-6的含量.结果 体积分数为4%终浓度的休克淋巴液作用6 h后,PMVECs中iNOS、TNF-α和IL-6的mRNA表达以及培养上清液中MDA、NO、TNF-α和IL-6水平均显著高于空白对照组、正常淋巴液组、休克血浆组、正常血浆组和无血清对照组;且休克血浆作用PMVEC 6 h后的iNOS、TNF-α和IL-6的mRNA表达及培养上清液中NO水平均显著高于空白对照组、正常淋巴液组、正常血浆组和无血清对照组(P<0.05或P<0.01).结论 4%终浓度的休克淋巴液可致大鼠PMVECs中iNOS、TNF-α和IL-6的mRNA表达增强,促进自由基释放,从而诱导细胞损伤.