Induction of apoptosis following blunt chest trauma

The cause for the high morbidity of blunt chest trauma is not fully understood. It is still unclear if and to what extent a second insult, e.g., apoptotic tissue damage initiated by the primary insult itself, may contribute to the development of serious complications. This study was done to elucidate whether a pulmonary contusion may induce programmed cell death. Sixty-four Wistar rats were evenly randomized to eight experimental groups: four sets were subjected to a standardized blast wave injury and sacrificed 6, 24, 48, and 72 h after the trauma; four groups served as controls for the same time points. Lung and liver samples were stained (H & E; TUNEL), and PMN infiltration was determined by myeloperoxidase (MPO) activity. Caspase 8 was analyzed by Western blot, and TNF-alpha plasma levels by ELISA. Postmortem examination revealed bilateral pulmonary contusion in trauma animals with higher (P < 0.05) numbers of apoptotic cells in lung but not in liver tissue as early as 6 h after the injury. This amount gradually increased and reached a maximum after 48 h: 6.8 +/- 1.1 apoptotic cells/hpf vs. 0.6 +/- 0.06 in controls. Chest trauma caused an increased expression of active caspase 8 in lung but not in liver tissue at 48 and 72 h. TNF-alpha plasma levels were not different. MPO activity in lung tissue of trauma animals increased (P < 0.05) after 6 h and peaked at 72 h. This study has provided the first evidence that apoptotic cell death in lung tissue is initiated following (experimental) pulmonary contusion. The exact mechanism remains, however, unclear and has to be elucidated further.     

Clinical assessment and radiograph following blunt chest trauma.

This study was undertaken to assess the accuracy of clinical examination in predicting significant injury following blunt chest trauma and to determine whether more selective use of frontal chest radiography could be achieved.     

Opposing effects of transmembrane and soluble Fas ligand expression on inflammation and tumor cell survival

Fas ligand (FasL) has been shown to mediate both apoptotic and inflammatory reactions. To rigorously assess the physiological role of different forms of the FasL molecule with regard to these two distinct processes, we isolated stably transfected lymphoma cell lines that expressed either murine wild-type FasL, membrane-only FasL, or functionally distinct forms of soluble FasL. First, the ability of these lines to induce an inflammatory response was assessed in vivo by injecting the transfectants intraperitoneally and measuring subsequent neutrophil extravasation into the peritoneal cavity. Second, lines were assessed by injecting the transfectants subcutaneously and monitoring their growth as solid tumors. Our study clearly demonstrated that the extent of inflammation induced by the transfectants directly correlated with their relative cytotoxic activities. A neutrophil response could only be elicited in mice with intact Fas death domains although Fas expression by the neutrophils was not essential. Lymphoma cells expressing the soluble FasL form corresponding to the natural cleavage product could not trigger apoptosis and did not induce a neutrophil response. In contrast to the other FasL transfectants, these cells survived as tumor transplants. However, expression of soluble FasL was not benign, but actually suppressed the inflammatory response and protected other transfectants from the effector mechanisms elicted by membrane-bound FasL.     

Traumatic tricuspid regurgitation following equine related blunt chest trauma and review of the literature

Myocardial injury following blunt chest trauma may be difficult to detect. We advocate for cardiac screening in such scenarios. Observation versus intervention should be based on symptoms and the degree of intracardiac disease.     

Divergent effects of activated neutrophils on inflammation, Kupffer cell/splenocyte activation, and lung injury following blunt chest trauma

Polymorphonuclear granulocytes (PMNs) have been attributed a primarily deleterious role in the pathogenesis of acute lung injury (ALI). However, evidence exists that PMNs might also act beneficially in certain types of ALI. In this regard, we investigated the role of activated neutrophils in the pathophysiology of lung contusion-induced ALI. We used the model of blunt chest trauma accompanied by PMN-depletion in male C3H/HeN mice. Animals received 25 μg/g body weight PMN-depleting antibody Gr-1 intravenously 48 h before trauma. Bronchoalveolar lavage (BAL) and lung tissue interleukin 6 (IL-6) were similarly elevated in PMN-depleted and control animals after trauma, whereas macrophage inflammatory protein 2 and monocyte chemoattractant protein 1 in BAL and lungs, IL-10 in BAL, and lung keratinocyte chemoattractant (KC) were even further increased in the absence of PMNs. Plasma IL-6 and KC were also increased in response to the insult and even further in the absence of PMNs. Chest trauma induced an enhanced release of IL-6, tumor necrosis factor α, macrophage inflammatory protein 2, monocyte chemoattractant protein 1, and IL-10 from isolated KU, which was blunted in the absence of PMNs. In the presence of PMNs, BAL protein was further increased at 30 h when compared with the 3-h time point, which was not the case in the absence of PMNs. Taken together, in response to lung trauma, activated neutrophils control inflammation including mediator release from distant immune cells but simultaneously mediate pulmonary tissue damage. Thus, keeping in mind potential inflammatory adverse effects, modulation of neutrophil activation or trafficking might be a reasonable therapeutic approach in chest trauma-induced lung injury.     

同种异体肾移植排斥患者血清sFas和sFasL水平的临床观察

目的　观察肾同种移植急性排斥患者血清可溶性Fas(sFas)和sFas配体(sFasL)的水平及临床意义。 方法　采用酶联免疫吸附试验(ELISA)分别对健康对照组及实验组透析前后sFas、sFasL进行检测。结果　对 照组sFas为(256.8±72.0)ng/L,sFasL为(227.9±65.9)ng/L;实验组透析前后sFas分别为(1225.7±467.6) ng/L、(1225.8±464.0)ng/L,sFasL分别为(227.9±147.2)ng/L、(226.9±109.6)ng/L。实验组与对照组的 sFas比较差异有显著性(P<0.01),而sFasL比较差异无显著性(P>0.05)。结论　sFas在排异的病理反应过 程中参与了细胞凋亡的抑制,透析并不能改善Fas FasL介导的细胞凋亡。     

同种异体肾移植排斥患者血清sFas和sFasL水平的临床观察

目的观察肾同种移植急性排斥患者血清可溶性Fas(sFas)和sFas配体(sFasL)的水平及临床意义.方法采用酶联免疫吸附试验(ELISA)分别对健康对照组及实验组透析前后sFas、sFasL进行检测.结果对照组sFas为(256.8±72.0)ng/L,sFasL为(227.9±65.9)ng/L;实验组透析前后sFas分别为(1 225.7±467.6)ng/L、(1 225.8±464.0)ng/L,sFasL分别为(227.9±147.2)ng/L、(226.9±109.6)ng/L.实验组与对照组的sFas比较差异有显著性(P＜0.01),而sFasL比较差异无显著性(P＞0.05).结论 sFas在排异的病理反应过程中参与了细胞凋亡的抑制,透析并不能改善Fas-FasL介导的细胞凋亡.     

Coronary artery occlusion following blunt chest trauma: a case report and review of the literature

Blunt chest trauma causing coronary artery occlusion and myocardial infarction is a rare but potentially fatal condition. We present the case of a healthy 29-year-old man who developed a myocardial infarction due to complete occlusion of the proximal right coronary artery following blunt chest trauma. A review of the literature found 63 cases of previously healthy patients under 40 years of age who developed coronary artery occlusion following blunt chest trauma; diagnosis in all cases had been proven by angiography or during autopsy. The presentation, results of electrocardiography and echocardiography and laboratory findings of these patients are described.     

不同程度阿尔茨海默病患者血清可溶性Fas及其配体水平的变化

目的探讨阿尔茨海默病(AD)患者血清可溶性Fas(s Fas)及其配体(s Fas L)水平的变化。方法选取AD患者100例,以54例老年疾病(包括糖尿病17例、高血压18例、高血脂15例、其它内分泌疾病4例)患者和46名健康体检者分别作为老年疾病对照组和正常对照组。采用酶联免疫吸附试验(ELISA)检测所有受检者血清s Fas、s Fas L水平,使用临床痴呆评定量表(CDR)、简易智能精神状态量表(MMSE)、Hachinski缺血指数量表(HIS)评定AD患者认知功能。结果 AD患者血清s Fas、s Fas L水平明显高于老年疾病对照组和正常对照组(P0.01);随着病程加重,其血清s Fas、s Fas L水平逐渐升高。AD患者血清s Fas与年龄、病程和MMSE评分呈正相关(r=0.856、0.834、0.799,P0.01),与Hachinski评分呈负相关(r=-0.714,P0.01);血清s Fas L与年龄、病程呈正相关(r=0.863、0.857,P0.01),与MMSE评分、HIS评分呈负相关(r=-0.801、-0.745,P0.01)。结论 AD患者存在血清s Fas、s Fas L水平异常,提示AD患者在发病过程中可能存在神经细胞凋亡,且血清s Fas、s Fas L水平可能与患者的智能水平、脑缺血程度有关。     

原位肝移植围手术期血浆可溶性Fas及FasL的变化研究

目的 观察原位肝移植围手术期血浆可溶性Fas(sFas)及可溶性FasL(sFasL)水平的变化.方法 20例终末期肝病患者,分别于麻醉前(T1)、无肝期15 min(T2)、再灌注期15 min(T3)、术后1 d(D1)4个时间点抽取桡动脉血,采用酶联免疫吸附法(ELISA)测定血浆sFas及sFasL水平.结果 从麻醉前到无肝期sFas及sFasL水平差异无统计学意义(P均>0.05),再灌注15 min sFas及sFasL水平较麻醉前均显著升高(P均＜0.05),术后1 d降至麻醉前水平.结论 sFas及sFasL参与了肝移植围手术期缺血/再灌注损伤.     

慢性乙型肝炎患者治疗前后血清sFas和sFasL的变化及其临床意义

目的　探讨各类慢性乙型肝炎(CHB)治疗前后血清可溶性Fas(sFas)和sFas配体(sFasL)的水平变 化及其意义。方法　用酶联免疫吸附试验(ELISA)检测118例各种类型CHB患者经前列腺素E1等药物治疗前 后血清sFas、sFasL的水平,并与30名健康献血者比较。结果　118例各种类型CHB患者的sFas明显高于正常 对照组(P<0.01、0.01、0.01、0.05),且重型肝炎(FH)>CHB重度>CHB中度>CHB轻度(P均<0.01),sFas 的升高与总胆红素(TBil)呈显著性正相关(r=0.605,P<0.01);FH和CHB重度的sFasL明显高于正常对照组 (P均<0.01)。恢复期各组sFas和sFasL均比治疗前明显降低(P均<0.01)。结论　血清sFas、sFasL的水平 与CHB病情密切相关,监测二者变化有助于判断病情和观察疗效。     

Functional expression of Fas and Fas ligand on human colonic intraepithelial T lymphocytes

The expression of Fas, a cell surface receptor directly responsible for triggering cell death by apoptosis, and its ligand (FasL) was investigated on both human colonic intraepithelial T lymphocytes (IELs) and peripheral blood mononuclear lymphocytes (PBMLs). FACS analysis indicated that IELs have increased expression of Fas compared with PBMLs, together with the progress activation marker, CD45RO. A discrete fraction of freshly isolated IELs also constitutively expressed FasL, perhaps as a result of recent in vivo activation. Using monoclonal antibody APO2.7, which detects mitochondrial 7A6 antigen specifically expressed by cells undergoing apoptosis, we further investigated the apoptosis-inducing effect of anti-Fas monoclonal antibody (CH11) on both IELs and PBMLs. FACS analysis revealed that CH11 increased the percentage of apoptotic cells, in IELs but not in PBMLs. Culture with anti-FasL monoclonal antibody (4H9) significantly recovered cell viability in IELs, but not in PBMLs. These results indicate that IELs constitutively express both Fas and FasL and that Fas crosslinking generates signals resulting in apoptosis, outlining a potential mechanism involved in intestinal tolerance.     

慢性乙型肝炎患者治疗前后血清sFas和sFasL的变化及其临床意义

目的探讨各类慢性乙型肝炎(CHB)治疗前后血清可溶性Fas(sFas)和sFas配体(sFasL)的水平变化及其意义.方法用酶联免疫吸附试验(ELISA)检测118例各种类型CHB患者经前列腺素E1等药物治疗前后血清sFas、sFasL的水平,并与30名健康献血者比较.结果 118例各种类型CHB患者的sFas明显高于正常对照组(P＜0.01、0.01、0.01、0.05),且重型肝炎(FH)＞CHB重度＞CHB中度＞CHB轻度(P均＜0.01),sFas的升高与总胆红素(TBil)呈显著性正相关(r=0.605,P＜0.01);FH和CHB重度的sFasL明显高于正常对照组(P均＜0.01).恢复期各组sFas和sFasL均比治疗前明显降低(P均＜0.01).结论血清sFas、sFasL的水平与CHB病情密切相关,监测二者变化有助于判断病情和观察疗效.     

糖皮质激素受体基因转染对肺泡巨噬细胞表达炎性因子的影响

目的:分析过量表达的外源性糖皮质激素受体在调控肺泡巨噬细胞表达炎性细胞因子中的作用。方法:实验于2005-10/2006-03在解放军南京军区南京总医院呼吸病实验室完成。①选用成年健康Wistar大鼠10只,雌雄不拘。采用大鼠糖皮质激素受体真核表达质粒pCMV-rGR转染体外培养肺泡巨噬细胞,将肺泡巨噬细胞分为转染组与非转染组,其中转染组细胞在质粒转染24h后用于实验,各组对以下时相点进行观察:正常对照(加等量生理盐水)、脂多糖 地塞米松作用4,8和12h,脂多糖与地塞米松作用浓度分别为100μg/L,1×10-5mol/L,于不同时相点收集细胞。②转染24h后WesternBlotting检测细胞总蛋白和核蛋白中糖皮质激素受体蛋白表达变化。同时收集细胞培养上清,采用酶联免疫吸附法浊定细胞培养上清中肿瘤坏死因子α、白细胞介素1β水平。③多组间比较用方差分析,两两比较用SNK检验。结果:①质粒转染肺泡巨噬细胞后糖皮质激素受体表达变化:转染组细胞总蛋白中糖皮质激素受体表达明显高于非转染组(3.75±0.25,1.21±0.16,P<0.01),转染组细胞核蛋白中糖皮质激素受体表达量与非转染组相近(1.15±0.12,1.09±0.11,P>0.05)。②脂多糖和地塞米松作用后核蛋白中糖皮质激素受体表达变化:脂多糖和地塞米松作用后4,8,12h转染组细胞核蛋白中糖皮质激素受体表达量分别为4.02±0.09,2.13±0.08,1.22±0.15,非转染组细胞核蛋白中糖皮质激素受体表达量分别为4.08±0.13,2.09±0.11,1.19±0.17,两组细胞核蛋白中糖皮质激素受体表达变化趋势一致,均于4h达到峰值,8h显著降低,12h达到最低,两组同时相点比较,差异不明显(P>0.05)。③脂多糖和地塞米松作用后细胞培养上清中细胞因子表达变化:转染组脂多糖和地塞米松作用后4,8,12h肿瘤坏死因子α表达分别为(32.16±3.99),(56.82±8.43),(73.34±9.97)ng/L,非转染细胞分别为(34.28±4.25),(59.82±9.65),(71.41±8.35)ng/L;转染组脂多糖和地塞米松作用后4,8,12h细胞上清中白细胞介素1β质量浓度分别为(67.57±18.37),(83.17±8.55),(97.55±8.27)ng/L,非转染分别为(62.23±15.64),(85.72±9.37),(99.07±9.25)ng/L。两组细胞上清中肿瘤坏死因子α、白细胞介素1β表达随时间逐渐增加,均于12h均达到峰值,同时相点两组比较均无明显差异(P>0.05)。结论:①通过体外糖皮质激素受体基因转染可以有效地在肺泡巨噬细胞中过量表达外源性糖皮质激素受体,并且定位于胞质内。②地塞米松作用后,肺泡巨噬细胞核内糖皮质激素受体表达明显增加,但随后又出现表达下调现象。③体外基因转染后胞质中过量表达的外源性糖皮质激素受体不能发生核移位,所检测的均为内源性糖皮质激素受体。④通过基因转染而过量表达的外源性糖皮质激素受体,不能发挥正常的抑炎生物学活性。     

Cardiopulmonary,histological, and inflammatory alterations after lung contusion in a novel mouse model of blunt chest trauma

Severe blunt chest trauma remains an important injury with high morbidity and mortality. However, the associated immunological alterations are poorly understood. Existing big animal models require large-scale settings, are often too expensive, and research products for immunological studies are limited. In this study we aimed to establish a new model of blunt, isolated and bilateral chest trauma in mice and to characterize its effects on physiological and inflammatory variables. Male C3H/HeN mice (n = 9-10/group) were anesthetized and a femoral artery was catheterized. The animals were subjected to trauma or sham procedure and monitored for 180 min. Blunt chest trauma was induced by a blast wave focused on the thorax. Trauma intensity was optimized by varying the exposure distance. Blood pressure, heart rate, respiratory rate, arterial blood gases and plasma cytokine levels were measured. Macroscopic and microscopic examinations were performed. In addition, outcome was evaluated in a 10-day survival study. Chest trauma caused a drop (P < 0.05) in blood pressure and heart rate, which partly recovered. Blood gases revealed hypoxemia and hypercarbia (P < 0.05) 180 min after trauma. There was marked damage to the lungs but none to abdominal organs. Histologically, the characteristic signs of a bilateral lung contusion with alveolar and intrabronchial hemorrhage were found. Plasma interleukin-6 and tumor necrosis factor alpha were considerably increased after 180 min. Blunt chest trauma resulted in an early mortality of 10% without subsequent death. On the basis of these findings, this novel mouse model of blunt chest trauma appears suitable for detailed studies on immunological effects of lung contusion.     

The pulmonary and hepatic immune microenvironment and its contribution to the early systemic inflammation following blunt chest trauma

OBJECTIVE: Blunt chest trauma is accompanied by an early increase in plasma cytokine concentrations. However, the local sources of these mediators are poorly defined. We investigated the impact of blunt chest trauma on the inflammatory mediator milieu in different compartments (lung tissue, bronchoalveolar lavage, liver tissue, Kupffer cells, plasma) along with the time course of trauma-induced pulmonary endothelial barrier dysfunction to elucidate potential relationships. In addition, the correlation between intratracheally instilled interleukin-6 and its systemic release were studied. DESIGN: Prospective, randomized, controlled animal study. SETTING: Basic science laboratory of a university affiliated level 1 trauma center. SUBJECTS: Male C3H/HeN mice, 8-9 wks old, n = 141. INTERVENTIONS: Blunt chest trauma induced by a focused blast wave, intravenous injection of Evans blue, and intratracheal instillation of recombinant human interleukin-6. MEASUREMENTS AND MAIN RESULTS: Two hours after blunt chest trauma, plasma interleukin-6 was markedly increased. Simultaneously, interleukin-6, tumor necrosis factor-alpha, macrophage inflammatory protein-2, monocyte chemotactic polypeptide-1 and neutrophil/monocyte accumulation in bronchoalveolar lavage and interleukin-6, monocyte chemotactic polypeptide-1, and myeloperoxidase activity in lung tissue were significantly increased. This was accompanied by a coinciding elevation in the Evans blue lung-plasma ratio. Recombinant human interleukin-6, instilled intratracheally before blunt chest trauma, was detected in a dose-dependent manner in the plasma of the mice. Additionally, Kupffer cell interleukin-6, tumor necrosis factor-alpha, and interleukin-10 production was significantly augmented as early as 30 mins after the insult. CONCLUSIONS: These results indicate that early increased cytokine concentrations in the lung, particularly interleukin-6, are important mediator sources as their local peak coincides with the systemic inflammatory response and is accompanied by a simultaneous impaired function of the pulmonary endothelial barrier. A direct relationship between their local and systemic concentrations can be established. Furthermore, this is the first study to show that Kupffer cells are activated early after blunt chest trauma.     

白细胞介素6对小鼠肺泡巨噬细胞CD14、清道夫受体表达的调节功能

目的:阐明白细胞介素(IL-6)对小鼠肺泡巨噬细胞(alveolarmacrophage,AM)清道夫受体(scavengerreceptor,SR)及CD14表达的直接调节作用及探讨IL-6提高细胞免疫功能的作用。方法:分离培养小鼠AM,以不同剂量(0,0.01,0.1,1,10,100μg/L)IL-6刺激细胞16h或以100μg/LIL-6在不同时间(0,2,4,8,12,16h)刺激细胞,采用免疫细胞化学及RT-PCR方法观察SR,CD14表达变化。结果:IL-6刺激AM能增强CD14蛋白表达并抑制SR蛋白表达,低至0.01μg/LIL-6刺激16h或100g/LIL-6刺激6h后就能显著增强CD14mRNA并明显抑制SRmRNA表达,与此同时,CD14蛋白表达也明显增强而SR蛋白表达显著下降。结论:IL-6刺激AM能在mRNA及蛋白水平显著增强CD14表达并抑制SR表达。