失血性休克鼠肺组织Toll样受体基因的表达

目的探讨单纯性失血性休克（无复苏）对肺组织中Toll样受体（TLR）mRNA表达的影响及意义。方法C57BL／6小鼠45只。随机分为失血组、脂多糖（LPS）组（阳性对照组。尾静脉注射LPS5mg／kg）、假手术组（阴性对照组）。每组15只。心脏穿刺复制失血性休克模型，在不同时间点取出肺组织。提取总RNA，通过逆转录-聚合酶链反应（RT—PCR）半定量方法检测肺组织TLR2 mRNA和TLR4 mRNA的表达水平。结果在失血性休克和LPS刺激后肺出现明显的中性粒细胞浸润、红细胞渗出。正常肺组织中有TLR2 mRNA、TLR4 mRNA表达；住失血性休克和LPS刺激后0、1、2、4和6h肺组织TLR2 mRNA、TLR4 mRNA表达均逐渐增加；而假手术组未发生明显改变。结论失血性休克后肺组织TLR2 mRNA、TLR4 mRNA表达增加与急性肺损伤（ALI）的发生有密切关系。除增强了机体非特异性免疫能力外，同时增加了宿主对随后各种刺激的易感性。过度表达的TLR2、TLR4可能造成组织、器官结构和功能损害。     

失血性休克小鼠心肌Toll样受体2/4 mRNA的表达

目的 探讨无复苏对失血性休克小鼠心肌Toll样受体（TLR）表达变化的影响及其意义。方法 将45只C57BL／6小鼠随机分为失血性休克模型组、假手术组、脂多糖（LPS）组（由尾静脉注射LPS5mg／kg），每组15只；采用心脏穿刺法建立小鼠失血性休克模型。心肌TLR2mRNA和TLR4mRNA表达采用逆转录-聚合酶链反应（RT—PCR）方法进行分析；测定左室收缩末压（LVESP）以反映左室收缩功能。结果 ①与假手术组比较，失血性休克及LPS刺激后小鼠的动脉血压出现下降，均可导致左室收缩功能障碍；②失血性休克及LPS刺激后TLR2和TLR4的mRNA表达水平均出现不同程度上调，而假手术组在各时间点未见明显改变。结论 ①失血性休克及LPS刺激后心肌TLR2及TLR4的mRNA表达上调与心功能障碍存在密切联系，但这两种病理状态下的信号转导通路可能存在差异；②失血性休克和LPS刺激后TLR2、TLR4的mRNA升高增强了机体的天然免疫功能，提高了机体对急性炎症的应激能力，对机体具有保护作用，但其过度表达也可能对组织、器官功能产生损害。     

Gender dimorphism in trauma-hemorrhage-induced thymocyte apoptosis.

Studies indicate that immune responses after trauma-hemorrhage are significantly depressed in males compared with enhanced immune responses in females under such conditions. Although androgen depletion in male mice by castration before soft tissue trauma and hemorrhagic shock prevents the depression of cell-mediated immunity, the underlying mechanism responsible for this remains unclear. Because the thymus is the primary location of T-cell lymphopoiesis and thymocytes express a large number of androgen receptors, we investigated whether differences in thymic apoptosis might contribute to the divergent immune response in males versus females after trauma-hemorrhage. To study this, male and female C3H/HeN mice were subjected to sham operation or soft tissue trauma (laparotomy) and hemorrhagic shock followed by fluid resuscitation. Animals were killed 72 h thereafter and thymocytes were isolated. Thymocyte interleukin 3 (IL-3) release was significantly suppressed in males, but not females, after trauma-hemorrhage. A parallel increase in thymic apoptosis that was primarily in the CD8+ thymocyte subset was observed in the males. Furthermore, in vitro treatment of thymocytes with 5a-dihydrotestosterone (DHT) increased the rate of apoptosis and decreased IL-3 release in a dose-dependent manner. Thus, the gender-dependent dimorphic immune response after trauma-hemorrhage may be in part due to an androgen-induced increase in thymic apoptosis in males under such conditions.     

Gender-related therapy: early IL-10 administration after hemorrhage restores immune function in males but not in females

Recent experimental studies have found gender differences in the immune response after hemorrhagic shock with an enhanced immune function and lower mortality after subsequent sepsis in females than in males. Interleukin-10 (IL-10) has been shown to play a potential role in the treatment of early proinflammatory state after hemorrhagic shock. Although studies showed beneficial effects of the treatment with IL-10, it remains unclear whether the effects are gender related. To study this, male and female CBA/J mice were subjected to hemorrhage (35+/-5 mmHg for 90 min and fluid resuscitation) or sham operation. At resuscitation, each received either recombinant murine IL-10 (rmIL-10) or placebo i.p. At 48 h after resuscitation, peritoneal macrophages (pMphi) and splenocytes were harvested. IL-1beta and IL-12 release by pMphi and splenocyte proliferation and splenocyte IL-2 and interferon (IFN)-gamma release capacity were assessed. Interleukin-10 plasma levels were not increased after rmIL-10 treatment. The results indicate that rmIL-10 treatment restores depressed immune response (splenocyte proliferation, IFN-gamma, IL-1beta in males after hemorrhagic shock. In contrast, the immune responses after shock in females were not influenced by rmIL-10, with the exception of depressed splenocyte proliferation. In addition, sham-operated male mice treated with rmIL-10 showed immune depression compared with the placebo group. Thus, administration of rmIL-10 during resuscitation after hemorrhage produces salutary effects on the depressed immune responses in males but did not further enhance the immune functions in females under those conditions.     

Preservation of splenic immune functions by female sex hormones after trauma-hemorrhage

OBJECTIVES: Immune functions are markedly depressed in males but not in proestrus females after trauma-hemorrhage. Nonetheless, it is unclear what role sex steroids play in the maintenance of immune function in females after trauma-hemorrhage. DESIGN: Prospective, controlled animal study. SETTING: University research laboratory. SUBJECTS: Eight-week-old female CBA/J mice. INTERVENTIONS: Mice underwent sham-ovariectomy or ovariectomy. Two weeks thereafter, ovariectomized and proestrus sham-ovariectomized mice were subjected to laparotomy (i.e., soft tissue trauma) and hemorrhagic shock (35 +/- 5 mm Hg for 90 mins, resuscitated) or sham operation. Splenocyte proliferation and interleukin-2, interleukin-3, and interferon-gamma release were determined at 2 hrs after trauma-hemorrhage. MEASUREMENTS AND MAIN RESULTS: These immune functional capacities were maintained in proestrus sham-ovariectomized mice after trauma-hemorrhage, whereas they were suppressed in ovariectomized mice subjected to trauma-hemorrhage. 17beta-Estradiol in vitro had no effect on splenocyte functions in proestrus sham-ovariectomized females; however, addition of 17beta-estradiol to splenocytes from ovariectomized females subjected to trauma-hemorrhage normalized immune functional capacities. CONCLUSIONS: These findings suggest that elevated circulating 17beta-estradiol in proestrus females plays a direct role in the maintenance of immunocompetence after trauma-hemorrhage.     

The effect of dehydroepiandrosterone on hemorrhage-induced suppression of cellular immune function

OBJECTIVE: To determine whether the steroid hormone dehydroepiandrosterone (DHEA) improves cellular immune functions after hemorrhagic shock. DESIGN AND SETTING: Prospective controlled study in a research laboratory at an university medical center. SUBJECTS: Male NMRI mice. INTERVENTIONS: Animals received 0.9% saline or DHEA (20 mg/kg subcutaneously) before induction of a volume-controlled hemorrhagic shock (55% of estimated circulating blood volume) by retro-orbital puncture. One hour after hemorrhage mice underwent fluid resuscitation by intravenous infusion of lactated Ringer's solution (300% of the shed blood). Separate groups of mice were killed to obtain whole blood and spleen 1 h after hemorrhage, 1 h after fluid resuscitation, and 24 h after hemorrhage to determine lymphocyte distribution (CD4(+), CD8(+), NK1.1-AG(+)), splenocyte apoptosis, and plasma concentrations of tumor necrosis factor-alpha and interleukin-10. MEASUREMENTS AND RESULTS: Hemorrhage in control mice was associated with a rapid increase in circulating NK cell numbers. Elevated splenocyte apoptosis, an increased CD4/CD8 ratio, and decreased number of circulating CD8(+) T-cells was observed 24 h after hemorrhagic shock. DHEA administration was accompanied by a normalization of splenocyte apoptosis and lymphocyte migration. Induction of hemorrhagic shock did not affect TNF-alpha or IL-10 plasma concentrations in either treatment group. CONCLUSIONS: DHEA administration improves cellular immune function after hemorrhage and may therefore be beneficial in patients with hemorrhagic shock.     

热休克蛋白70对严重创伤休克后肝脏的作用

目的 探讨热休克蛋白70（HSP70）对严重创伤休克后肝脏作用.方法 成年Wistar大鼠,采用双侧股骨骨折伴失血性休克致严重创伤模型,动态观察伤后8 h大鼠肝组织HSP70、血清肝功能生化指标、肝脏病理等变化.HSP70表达测定采用免疫印迹法,并进行计算机图像分析.结果 伤后HSP70在肝组织中表达迅速增加,6 h达到峰值,伤后8 h仍维持较高水平;创伤合并休克后,HSP70表达高峰提前至伤后4 h,持续表达至伤后6 h后逐渐下降,死亡前在肝组织中仍有少量表达.创伤休克后血清ALT、TB伤后4 h开始明显增高（P＜0.01）,白蛋白下降（P＜0.01）.肝脏镜下创伤休克后6 h肝窦内出现较多炎性细胞浸润.结论 在创伤休克早期,HSP70可能参与了肝组织细胞抗损伤机制的启动,但随着休克时间的延长,HSP70的过高持续表达,则可能对肝脏造成损害.HSP70在创伤休克后肝保护与肝损害过程中可能发挥双重作用.     

新型钙增敏剂哒嗪酮对低血容量性休克大鼠肝脏结构与功能的影响

目的：研究哒嗪酮（ＭＣＩ１５４）对低血容量性休克大鼠肝脏功能和结构改变的影响。方法：用大鼠低血容量性休克模型,设立ＭＣＩ１５４治疗组和生理盐水对照组,测定肝脏组织血流量以及肝脏组织提取液和线粒体悬液的生化指标,观察肝脏结构及超微结构的变化。结果：ＭＣＩ１５４治疗后肝脏组织血流量和Ｈ＋ＡＴＰ酶的活性显著高于相应的生理盐水对照组,组织提取液中丙氨酸转氨酶、乳酸脱氢酶、β葡萄糖醛酸酶和丙二醛以及线粒体内钙含量等指标显著低于相应的生理盐水对照组,肝脏细胞结构、超微结构损伤程度明显轻于生理盐水对照组。结论：ＭＣＩ１５４可显著增加大鼠肝脏组织血流量,减轻肝细胞损伤程度,延缓线粒体内钙超载进程,保护Ｈ＋ＡＴＰ酶的活性,对低血容量性休克大鼠肝脏结构和功能具有显著的保护作用。     

Prevention of age-related T cell apoptosis defect in CD2-fas-transgenic mice

T cell dysfunction and thymic involution are major immunologic abnormalities associated with aging. Fas (CD95) is a bifunctional molecule that is critical for apoptosis and stimulation during T cell development, but the role of Fas during aging has not been determined. Fas expression and function on T cells from old (22-26-mo-old) mice was compared with young (2-mo-old) mice and old CD2-fas-transgenic mice. Fas expression and ligand-induced apoptosis were decreased on T cells from old mice compared with young mice. This correlated with an age- related increase in CD44+Fas- T cells. There was a marked decrease in the proliferation of T cells from old mice after anti-CD3 stimulation compared with young mice. Anti-CD3-stimulated T cells from young mice exhibited increased production of interleukin (IL)-2 and decreased production of interferon-gamma and IL-10 compared with old mice. There was an age-related decrease in the total thymocyte count from 127 +/- 10 cells in young mice compared with 26 +/- 8 x 10(6) in old mice. In 26-mo-old CD2-fas-transgenic mice, Fas and CD44 expression, Fas-induced apoptosis, T cell proliferation, and cytokine production were comparable to that of the young mice. These results suggest that T cell senescence with age is associated with defective apoptosis, and that the CD2-fas transgene allows maintenance of Fas apoptosis function and T cell function in aged mice comparable to that of young mice.     

Endothelial cell activation in emergency department patients with sepsis-related and non-sepsis-related hypotension

Previous studies found increased circulating levels of biomarkers related to endothelial cell activation in patients with sepsis, particularly in the most severe sepsis stages of sepsis shock. It remains unclear, however, whether this activation is mainly driven by sepsis-specific mechanisms or occurs as a generalized inflammatory response. The objective of this analysis was to compare patterns of biomarkers of endothelial cell activation in patients with hypotension due to sepsis and nonsepsis etiologies. This is a secondary analysis of a prospective, observational cohort study including emergency department patients older than17 years with an episode of hypotension defined as any systolic blood pressure measurement less than 100 mmHg. Etiology of hypotension episodes was classified as sepsis or nonsepsis (eg, cardiac or hemorrhagic). Endothelial activation biomarkers of cell adhesion (E-selectin, vascular cell adhesion molecule 1 [VCAM-1], and intercellular adhesion molecule 1 [ICAM-1]), coagulation (plasminogen activator inhibitor 1 [PAI-1]), and vascular endothelial growth factor (VEGF) signaling (VEGF, soluble fms-like tyrosine kinase 1 [sFLT-1]) were assayed. A total of 161 patients were analyzed. Hypotension was classified as sepsis (n = 69), nonsepsis (cardiac [n = 35], hemorrhagic [n = 12]), or indeterminate (n = 45). With the exception of PAI-1, median plasma levels of all endothelial markers were significantly higher in patients with sepsis compared with nonsepsis etiology (P < 0.05 for all comparisons). Logistic regression analysis, adjusted for age, sex, mean blood pressure level, and mortality, confirmed a significant association of E-selectin (odds ratio [OR], 3.7; 95% confidence interval [CI], 1.7-7.8, P < 0.001) and sFLT-1 (OR, 2.0; CI, 1.1-3.8; P < 0.03) with sepsis etiology. Biomarkers VCAM-1 (OR, 2.0; CI, 0.88-4.4; P = 0.1), VEGF (OR, 1.5; CI, 0.98-2.2; P = 0.06), ICAM-1 (OR, 1.5; CI, 0.9-2.6; P = 0.2), and PAI-1 (OR, 1.4; CI, 0.8-2.3; P = 0.2) did not reach statistical significance. This study found a sepsis-specific activation of endothelium activation markers, particularly E-selectin and sFLT-1, in emergency department patients with hypotension.     

Autonomic receptors in urinary tract: sex and age differences

As age and sex affect the function of the lower urinary tract, we studied the characteristics of adrenergic and cholinergic receptors in various parts of lower urinary tract smooth muscle of young (6 months) and old (4 1/2-5 years) male and female rabbits. Saturation experiments performed with [3H]prazosin, [3H]yohimbine, [3H]dihydroalprenolol and [3H]quinuclidinyl benzylate in rabbit bladder base, bladder dome and urethra indicate the presence of regional, sex- and age-related differences in the density of alpha-1, alpha-2, and beta adrenergic and muscarinic cholinergic receptors. Alpha-2 adrenergic receptor density is considerably higher in the female than in the male urethra of both age groups, whereas the higher density of beta adrenergic receptors in the female than in the male bladder base is observed only in the younger animals. The density of muscarinic receptors is higher in bladder dome than in bladder base or urethra in young rabbits of both sexes. In the old animals, the density of muscarinic receptors in bladder base increases to the level observed in bladder dome. Inhibition experiments with selective adrenergic agonists and antagonists indicate that the pharmacological profiles of alpha-2 adrenergic receptors in the urethra and beta adrenergic receptors in the bladder dome and bladder base are similar in both sexes and at both ages. Beta-2 adrenergic receptors are shown to be predominant in bladder base and bladder dome of rabbits. Parallel studies in rabbit urethra, adult rat cortex and neonatal rat lung show that the urethral alpha-2 adrenergic receptors are of the alpha-2A subtype.     

Differential fluid regulation during and after soft tissue trauma and hemorrhagic shock in males and proestrus females

Gender differences in immune and organ functions have been described in different rodent models of trauma- and pressure-controlled hemorrhagic shock. We hypothesized that gender influences the regulation of plasma and tissue fluids in rats under such conditions. To study this we used male and weight matched proestrus female Sprague-Dawley rats, which were assigned to three groups (n = 7/group): sham, maximal bleedout (trauma and 45 min of blood pressure at 35 mmHg without resuscitation), or 5 h after completion of trauma-hemorrhage and resuscitation. Trauma-hemorrhage involved midline laparotomy and approx. 90 min of hemorrhagic shock (35 mmHg), followed by fluid resuscitation (4x the shed blood volume with Ringers lactate). (51)Cr-EDTA, (125)I-albumin distribution, and wet weight/dry weight were used to calculate plasma volume and extracellular fluid volume and cellular water content. Proestrus female rats showed significantly higher plasma volumes compared with weight-matched males. The volume of blood withdrawn in the first 15 min of hemorrhagic shock was significantly less in proestrus females compared with males; however, there was no significant difference in the total shed blood volume. Moreover, proestrus females showed less interstitial edema formation compared with male rats at 5 h after resuscitation. We conclude that differences in the regulation of plasma and tissue volumes exist between males and proestrus females during and after trauma-hemorrhage. The increased circulating blood volume could contribute the improved immune and organ functions in proestrus females under those conditions.     

Early interleukin-10 treatment improves survival and enhances immune function only in males after hemorrhage and subsequent sepsis

Recent studies have demonstrated gender differences in the immune response following hemorrhagic shock with an enhanced immune function and lower mortality following subsequent sepsis in females. Early interleukin-10 (IL-10) treatment has been shown to have beneficial effects on the depressed immune function in males, but not in females following shock. However, it remains unclear if the observed gender-related effect of IL-10 treatment results in an advantage following subsequent polymicrobial sepsis. To study this, male and female CBA/J mice (age 2-3 months) were subjected to hemorrhage (35 +/- 5 mmHg for 90 min and fluid resuscitation). At resuscitation, each received either 10 microg of recombinant murine IL-10 or placebo i.p.. At 48 h after resuscitation, either peritoneal macrophages (pMphi) and plasma were harvested, or polymicrobial sepsis was induced by cecal ligation and puncture (CLP). Following CLP, either survival over 10 days was measured, or pMphi and plasma were harvested 4 h after CLP to assess TNF-alpha, IL-6, IL-10, and prostaglandin E2 (PGE2) release of pMphi and plasma levels of IL-10, free testosteron, and 17-beta estradiol. Early IL-10 treatment restored depressed proinflammatory immune response in males (TNF-alpha and PGE2), which was associated with an enhanced survival (P < 0.05) following subsequent sepsis as compared with placebo-treated mice (8/20 and 1/20, respectively). In contrast, the immune response and survival in females receiving IL-10 was not significantly changed, although females treated with IL-10 had a trend towards higher mortality (7/15 and 2/15, respectively; P = 0.08). Thus, early IL-10 anti-inflammatory treatment following hemorrhage has potential beneficial effects only in males associated with enhanced survival following subsequent sepsis.     

液体复苏治疗失血性休克疗效的系统评价

目的：评价限制性液体复苏与常规液体复苏治疗失血性休克的疗效。方法：计算机检索英文数据库：PubMed、Science Direct、EBSCO Host、Embase、The Cochrane Library,中文数据库：中国知网、万方数据库、维普数据库,检索年限：1991-01-01-2013-08-01。收集限制性液体复苏与常规液体复苏治疗失血性休克的随机对照试验,对纳入研究进行质量评价后,采用RevMan 5.0软件进行Meta分析。结果：18个RCT均为低质量文献,Meta分析结果显示：限制性液体复苏与常规性液体复苏比较,可显著改善失血性休克患者血清乳酸[MD=-0.59,95%CI（-0.79,-0.40）,P〈0.01]、血小板计数[MD=61.73,95%CI（55.61,67.85）,P〈0.01]、凝血酶原时间[MD=-4.00,95%CI（-4.26,-3.74）,P〈0.01]、血红蛋白[MD=12.16,95%CI（8.80,15.52）,P〈0.01],但血气碱剩余未见显著性差异。结论：限制性液体复苏较常规液体复苏治疗失血性休克患者疗效更好。     

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.     

Dependence of liver injury after hemorrhage/resuscitation in mice on NADPH oxidase-derived superoxide

Hemorrhagic shock and resuscitation cause hepatocellular damage by mechanisms involving oxidative stress. However, the sources of free radicals mediating hepatocellular injury remain controversial. Thus, this study tested the hypothesis that NADPH oxidase plays a role in producing hepatocellular injury after hemorrhagic shock and resuscitation. Both wild-type and NADPH oxidase-deficient mice (p47(phox) knockout mice) were subjected to hemorrhagic shock (3 h at 30 mmHg). The mice were resuscitated over 30 min with the shed blood and additional lactated Ringer's solution (50% of the shed blood volume). Serum alanine aminotransferase (ALT) levels increased at 1 and 6 h postresuscitation in wild-type animals to 4735 +/- 1017 IU/L and 1450 +/- 275 IU/L (mean +/- SE), respectively, whereas in knockout mice, this ALT increase was blunted at both time points (732 +/- 241 IU/L and 328 +/- 69 IU/L, P < 0.05). Liver necrosis assessed histologically 6 h after the end of reperfusion was also attenuated in the knockout mice (3.5% +/- 0.95% of area vs. 0.9% +/- 0.26%, P < 0.05). In hemorrhaged wild-type mice, infiltrating neutrophils were twice as numerous compared with hemorrhaged NADPH oxidase-deficient animals 6 h after reperfusion. In knockout animals, hepatic 4-hydroxynonenal content, indicative of lipid peroxidation from reactive oxygen species, was blunted (6.7% +/- 0.6% vs. 26.4% +/- 2.3% of stained area, P < 0.05), as shown by immunohistochemistry. Immunohistochemical staining for 3-nitrotyrosine, indicative of reactive nitrogen species formation, was also blunted in the livers of knockout mice (11.6% +/- 2.8% vs. 37.4% +/- 3.4, P < 0.05). In conclusion, hemorrhagic shock and resuscitation cause hepatocellular damage via NADPH oxidase-mediated oxidative stress. The absence of NADPH oxidase substantially attenuates hepatocellular injury after hemorrhagic shock and resuscitation, blunts neutrophil infiltration, and decreases formation of reactive oxygen and reactive nitrogen species.     

Early up-regulation of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression in rats with hemorrhagic shock and resuscitation.

This study evaluated the effect of resuscitation fluids on intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Sprague-Dawley rats (n = 36) were subjected to a 27 mL/kg hemorrhage over 5 min followed by a 1 h shock and 1 h resuscitation. Animals groups included: 1) cannulation only (Sham); 2) hemorrhage only (NR); 3) resuscitation with 1:1 shed blood (Blood); 4) resuscitation with 3:1 lactated Ringer's (81 mL/kg, 3LR+); 5) no hemorrhage but infusion with 3:1 lactated Ringer's (3LR); and 6) resuscitation with .36:1 hypertonic saline (7.5%, 9.7 mL/kg, HTS). At the end of resuscitation, the spleen and lung were harvested for detection of adhesion molecule mRNA and protein by RT-PCR and immunostaining. ICAM-1 and VCAM-1 expression exhibited the following pattern: 3LR+ > HTS approximate to 3LR > Blood approximate to NR approximate to Sham. VCAM-1 mRNA in the lung of the 3LR+ group was 2 or more times more than the groups of Sham, NR, Blood, and 3LR (p < .05). ICAM-1 and VCAM-1 mRNA in the spleen was significantly increased in the 3LR+ group compared with the groups of Sham, NR, and Blood (p < .05). Animals in the 3LR+ group showed enhanced staining for ICAM-1 in the pulmonary microvessels and in the marginal and trabecular areas of the spleen. Pulmonary edema and inflammatory cell infiltration were observed only in the 3LR+ group. In summary, resuscitation with LR following hemorrhagic shock induced immediate up-regulation of ICAM-1 and VCAM-1, which was associated with tissue injury. Thus, the type of resuscitation fluid used affected resuscitation injury.     

Hypertonic saline resuscitation reduces apoptosis and tissue damage of the small intestine in a mouse model of hemorrhagic shock

The effect of hypertonic saline resuscitation on intestinal damage and the incidence of apoptosis after hemorrhagic shock were investigated. After anesthesia, male BALB/c mice weighing 24-34 g were hemorrhaged to the mean arterial pressure of 40 +/- 5 mmHg for 90 min. Animals were randomly assigned to four groups: 1) resuscitation with 4 mL/kg of 7.5% NaCl (hypertonic saline; HS) + shed blood (SB); 2) resuscitation with two times the volume of shed blood of lactated Ringer's solution (2LR) + SB; 3) sham (catheter only); or 4) control (no treatment). Intestinal damage was graded based on the extent of the vacuolation at the basal area of the intestinal villi. Apoptosis of the small intestines was examined with the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling method and with DNA laddering. Caspase-3 activation, heat shock protein (HSP) 70, and HSP40 were assessed by western blotting. Apoptosis of the small intestine and intestinal damage were significantly lower (P < 0.01) in the HS+SB group compared with the 2LR+SB group 2 h and 6 h after hemorrhagic shock and resuscitation, respectively. This corresponded with more DNA fragmentation in the small intestine of the 2LR+SB group compared with the HS+SB group 2 h after hemorrhage and resuscitation. In addition, we observed less caspase-3 activation in the small intestine of the HS+SB group compared with the 2LR+SB group at 2 h after resuscitation. The content of HSP40 and HSP70 in the HS+SB group was similar to that in controls, but slightly decreased in the 2LR+SB group. HS resuscitation reduced intestinal damage and apoptosis after hemorrhagic shock, suggesting that HS resuscitation may improve the outcome after hemorrhagic shock by reducing apoptosis and damage to the small intestine.