骨髓间充质干细胞移植对烟雾吸入性损伤兔早期肺组织的影响

目的 探讨骨髓间充质干细胞(MSCs)移植对烟雾吸入性损伤兔早期肺组织损伤的影响.方法 采用直接贴壁法体外培养兔MSCs,用流式细胞术鉴定MSCs.将48只兔制备烟雾吸入性损伤模型后按随机数字表法均分成致伤组和MSCs组.MSCs组伤后立即静脉给予含1×107个/ml MSCs的磷酸盐缓冲液(PBS)10 ml;致伤组则给予10 ml PBS.分别于干预后2、6和24 h活杀8只兔,取肺组织行组织病理学观察,同时进行肺损伤评分.结果 流式细胞术检测显示所培养细胞为MSCs.肺大体标本和光镜下均观察到MSCs组肺损伤程度较致伤组明显减轻.虽然MSCs组和致伤组伤后2 h肺损伤评分(分)比较差异无统计学意义(4.0±0.7比4.5±0.6,P＞0.05),但MSCs组伤后6 h和24 h肺损伤评分(分)明显低于致伤组(6 h:6.1±0.9比8.2±0.9,24 h:4.6±0.9比10.4±0.8,均P＜0.01).结论 MSCs移植能明显减轻烟雾吸入性损伤兔肺组织损伤,改善肺损伤评分.

Abstract：

Objective To explore the effect of bone marrow mesenchymal stem cells (MSCs)engraftment on lung tissue at early stage of smoke inhalation injury in rabbits. Methods MSCs were proliferated by the method of whole marrow culture and identified by flow cytometry. Forty-eight rabbits were randomly divided into smoke inhalation group (S group) and MSCs group (M group) after reproduction of rabbit smoke inhalation injury model. 10 ml of phosphate buffer saline (PBS) containing 1 × 107/ml MSCs was intravenously injected in M group, meanwhile 10 ml PBS was injected intravenously in S group. Eight rabbits were sacrificed at 2, 6 and 24 hours after intervention, and the lung tissue was harvested for morphological and pathological observation, and lung injury score was used to evaluate smoke inhalation injury.Results Cultured cells were confirmed to be MSCs with flow cytometry. Lung injury in rabbits of M group was less serious in morphology and histopathology than that in S group. Though there was no significance in lung injury score between M group and S group at 2 hours after injury (4.0±0.7 vs. 4.5±0.6, P＞0.05),the lung injury scores in M group at 6 hours and 24 hours after injury were significantly lower than those in S group (6 hours: 6.1±0.9 vs. 8.2±0.9, 24 hours: 4. 6±0.9 vs. 10.4±0. 8, both P＜0. 01). Conclusion Intravenous engraftment of MSCs could ameliorate lung injury induced by smoke inhalation, and improve lung injury score significantly.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

目的 初步探讨慢性血吸虫(SJ)感染对脓毒症小鼠的保护作用及其机制.方法 选择BALB/c雄性小鼠,按随机数字表法分组进行三部分实验.实验1:经腹部皮肤接种SJ尾蚴感染8周建立慢性SJ感染模型,分为正常组和SJ组,每组10只;用酶联免疫吸附法(ELISA)检测血清白细胞介素(IL-4和IL-10)、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)水平,实时荧光定量聚合酶链反应(PCR)检测腹腔巨噬细胞IL-10和TNF-α的mRNA表达,了解慢性SJ感染小鼠免疫状态.实验2:以脂多糖(LPS)腹腔注射诱导小鼠脓毒症模型,分为LPS组和SJ-LPS组,每组15只;用ELISA法动态观察注射LPS后0、24、48和72 h细胞因子的变化,0 h的水平相当于正常小鼠和SJ感染8周水平,观察慢性SJ感染对脓毒症过程的影响.实验3:分别以盲肠结扎穿孔术(CLP)和LPS诱导两种不同的脓毒症模型,评价慢性SJ感染对脓毒症小鼠72 h存活率的影响.结果 实验1:SJ组血清抗炎因子IL-4[(151.35±12.24)ng/L]和IL-10[(133.22±11.09)ng/L]水平较正常组[IL-4(56.32±8.66)ng/L,IL-10(48.17±7.23)ng/L]显著升高(均P＜0.05),并可使巨噬细胞向替代活化性巨噬细胞分化,慢性SJ感染使腹腔巨噬细胞高表达IL-10 mRNA(SJ组4.46±1.82,正常组1.52±0.60),抑制TNF-α mRNA表达(SJ组1.61±0.93,正常组2.32±1.03,均P＜0.05).实验2、3:慢性SJ感染小鼠血清IL-4、IL-10于注射LPS后0 h即显著升高,随后下降,至72 h仍明显高于LPS组[IL-4(ng/L):92.2±7.6比41.5±4.5;IL-10(ng/L):92.1±7.8比35.6±4.0,均P＜0.05];TNF-α、IFN-γ均于24 h达峰值后逐渐下降,至72 h SJ-LPS组仍显著低于LPS组[TNF-α(ng/L):82.9±5.6比91.5±5.2;IFN-γ(ng/L):44.1±4.8比52.6±4.0,均P＜0.05].慢性SJ感染可明显改善CLP或LPS所致脓毒症小鼠的存活率(CLP:80%比20%,LPS:70%比30%,均P＜0.05).结论 慢性SJ感染可使脓毒症小鼠血清抗炎因子升高,存活率上升,从而起到保护作用.

Abstract：

Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

胸部爆震伤致兔急性呼吸窘迫综合征模型的建立及相关因素分析

目的 构建室内胸部爆震伤致兔急性呼吸窘迫综合征(ARDS)模型并分析其发生机制及早期死亡原因,为研究肺爆震伤早期预警体系和治疗方法提供依据.方法 按照不同炸药量和致伤距离所产生的压强,将60只新西兰大白兔按随机数字表法分为5个致伤组和1个无致伤对照组.伤后观察存活率和组织病理学,并监测病理生理学指标、肺含水量.结果 冲击波压强低于1 210.5 mm Hg(1 mm Hg=0.133 kPa,A、B组)时,肺损伤较轻,表现为点状肺挫伤,肺简明损伤评定分级法(ALS)均在2级内,动物伤后24 h内全部恢复,长期存活无并发症.冲击波压强高于2 036.1 mm Hg(D、E组)时,肺损伤过重,表现为广泛的肺挫伤、肺门撕裂伤和肺内大血肿,AIS均大于5级,动物于伤后1 h内全部死亡.冲击波压强为1 917.3 mm Hg(C组)时,肺表现为广泛而恒定的挫伤,累及4个肺叶以上,AIS 4～5级,伤后6 h内出现动脉氧分压下降;肺组织可见肺泡壁水肿,部分肺泡壁断裂,肺泡融合;肺泡内充满大量炎性细胞,偶见透明膜形成.与对照组比较,C组兔致伤6 h肺湿/干重比值即显著升高(6.46±0.24比3.98±0.19,P＜0.01),血浆及支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)即明显升高[血浆TNF-α(ng/L):328.89±6.26比62.12±2.98,BALF TNF-α(ng/L):164.87±4.59比29.51±1.12;血浆IL-6(ng/L):128.51±4.13比19.32±1.53,BALF IL-6(ng/L):94.97±1.14比22.72±0.19,均P＜0.05].结论 在1 917.3 mm Hg爆炸压强的密闭环境下,冲击伤可诱导兔发生ARDS;TNF-α及IL-6参与爆震伤致ARDS的形成与发展;特定环境下,肺脏破裂致气胸为早期死亡原因,而冲击波致循环系统功能紊乱也是引起早期死亡的重要原因.

Abstract：

Objective To reproduce acute respiratory distress syndrome (ARDS) model in rabbit induced by chest blast injury and to analyze the pathogenesis and causes of early death in order to provide the basis for the early diagnosis of lung blast injury and its early-warning system to facilitate an early treatment.Methods Sixty healthy New Zealand white rabbits were divided into six groups according to the different explosion distance with the random number table method. The survival rate and its resulting pathological changes were observed and patho-physiological indexes and lung fluid content were determined at sequential time points post-explosion. Results Shock wave pressure less than 1 210. 5 mm Hg (1 mm Hg=0. 133 kPa,group A, B) resulted in limited injury to the lung within grade-2 as assessed with the abbreviated injury scale (AIS). The rabbits in these groups recovered soon and survived without any complication. Shock pressure higher than 2 036. 1 mm Hg (group D, E) caused severe injuries to the lung, including deep laceration, disruption of lung hilus and large hematoma in the lung, and the injury severity of lungs was assessed above grade-5 as assessed with AIS. All rabbits died within 1 hour post-explosion. The groups described above failed to meet the demand of an ARDS model for the present study. Shock wave pressure at 1 917. 3 mm Hg (group C) produced extensive contusion from grade-4 to grade-5 as assessed with AIS. The rabbits survived in poor general condition, and arterial partial pressure of oxygen (PaO2) lowered within 6 hours. Pathological examination showed extensive and constant multi-focal bleeding involving more than four lobes. The alveolar wall was edematous, with partial rupture and alveolar fusion in lung tissues was observed in the group C. Alveoli were filled with inflammatory cells, and hyaline membrane was formed occasionally. Compared with control group, the wet to dry weight ratio (W/D) in lungs increased obviously (6.46±0. 24 vs. 3. 98±0. 19, P＜ 0. 01) in group C within 6 hours postinjury. The contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in plasma and bronchoalveolar lavage fluid (BALF) were also increased distinctly compared with the control group [TNF-α (ng/L) in plasma: 328. 89± 6.26 vs.62.12±2. 98, TNF-α (ng/L) in BALF: 164.87±4.59 vs. 29. 51±1.12; IL-6 (ng/L) in plasma: 128. 51±4.13 vs. 19.32±1.53: IL-6 (ng/L) in BALF: 94.97±1.14 vs. 22.72±0. 19, all P＜0. 05]. Conclusion In an airtight environment, rabbit ARDS model can be reproduced successfully by blast injury with 1 917.3 mm Hg explosion pressure; TNF-α and IL-6 are involved in the pathogenesis and development of ARDS in blast injury. Pneumothorax as a result of lung rupture is the chief reason for early death and dysfunction of circulatory system is also an important reason in producing early death.     

谷氨酰胺对内毒素血症大鼠肠道损伤的保护作用及对血红素加氧酶-1表达的影响

目的 探讨谷氨酰胺(Glu)对内毒素血症大鼠肠道损伤的保护作用以及对血红素加氧酶-1(HO-1)表达的影响.方法 按随机数字表法将32只雄性SD 大鼠分为正常对照组、模型组、Glu组和Glu+锌原卟啉(ZnPP)组,每组8只.腹腔注射内毒素脂多糖(LPS)10 mg/kg制备内毒素血症动物模型.Glu组注射LPS前12 h灌胃Glu 1 g/kg;Glu+ZnPP组注射LPS前12 h灌胃Glu 1 g/kg,注射LPS前1 h静脉注射ZnPP 10 mmol/kg.术后12 h取回肠组织,测定髓过氧化物酶(MPO)活性及肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)含量,并进行肠组织学评分;用免疫组化法检测回肠组织HO-1表达.结果 与正常对照组比较,模型组回肠组织学评分、MPO活性及TNF-α、IL-10含量显著升高[组织学评分(分):3.3±0.4比1.1±0.6,MPO活性(U/g):0.40±0.08比0.26±0.07,TNF-α含量(ng/g):25.2±6.9比6.5±2.8,IL-10含量(ng/g):27.6±10.2比5.7±2.9,均P<0.01];HO-1表达较低.与模型组比较,Glu组组织学评分、MPO活性和TNF-α含量明显减低[组织学评分(分):1.6±0.5比3.3±0.4,MPO活性(U/g):0.25±0.05比0.40±0.08,TNF-α含量(ng/g):13.4±3.2比25.2±6.9,均P<0.01],IL-10含量(ng/g)显著升高(47.3±5.5比27.6±10.2,P<0.01),HO-1表达明显增加.Glu+ZnPP组与模型组各指标比较差异无统计学意义.结论 Glu能明显增强内毒素血症大鼠肠组织HO-1表达,明显减轻肠道炎症反应,从而保护肠黏膜.

Abstract：

Objective To investigate the protective effect of glutamine (Glu) pretreatment on intestinal injury induced by endotoxin and expression of heme oxygenase-1 (HO-1) in rats.Methods Thirty-two male Sprague-Dawley (SD) rats were randomly divided into four groups (n=8 in each group): normal control group, model group, Glu group and Glu+zinc protoporphyrin (ZnPP) group. In model group, endotoxemia was produced by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg). In Glu group, the rats received intragastraiclly 1 g/kg of Glu 12 hours before LPS intraperitoneal injection. In Glu+ZnPP group, the rats received 1 g/kg of Glu by gavage 12 hours before LPS intraperitoneal injection and ZnPP 10 mmol/kg intravenously via tail vein 1 hour before LPS injection. The distal ileum was harvested in full thickness 12 hours after LPS injection. The myeloperoxidase (MPO) activity, tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in the intestine were determined, the pathologic changes were observed and expressed in Chiu grade. The expression of HO-1 was evaluated by immunohistochemistry method. Results Compared with normal control group, the Chiu grade, MPO activity, the content of TNF-α and IL-10 were significantly increased in model group [Chiu grade: 3.3±0.4 vs. 1.1±0.6, MPO activity (U/g): 0.40±0.08 vs. 0.26±0.07, TNF-α (ng/g): 25.2±6.9 vs. 6.5±2.8, IL-10 (ng/g): 27.6±10.2 vs. 5.7±2.9, all P<0.01], and the expression of HO-1 was decreased. Compared with model group, the Chiu grade, MPO activity, the content of TNF-α in Glu group were significantly decreased [Chiu grade: 1.6±0.5 vs. 3.3±0.4, MPO activity (U/g): 0.25±0.05 vs. 0.40±0.08, the content of TNF-α (ng/g): 13.4±3.2 vs. 25.2±6.9, all P<0.01], while the level of IL-10 (ng/g) elevated (47.3±5.5 vs. 27.6±10.2, P<0.01), and the expression of HO-1 was increased. There was no difference in above mentioned indexes between model group and Glu+ZnPP group. Conclusion Glu pretreatment significantly ameliorates the expression of HO-1 of intestinal tissue induced by LPS in rats, and intestinal mucosa is protected with alleviation of inflammatory reaction in intestinal tract.     

普通肝素雾化吸入对内毒素性肺损伤大鼠肺泡局部凝血及炎症反应的影响

目的 观察内毒素诱导急性肺损伤(ALI)大鼠雾化吸入普通肝素(UFH)后的肺部局部效应,探讨其对肺泡内凝血、纤溶和炎症反应的作用.方法 87只雄性Wistar大鼠按随机数字表法分为假损伤组、模型组、肝素治疗组(HT组)和肝素预防组(HP组).静脉注射(静注)内毒素脂多糖(LPS)制备ALI模型.HP组和HT组分别于注射LPS前后给予UFH雾化吸入,模型组和假损伤组则雾化吸入生理盐水.各组分别于静注LPS后6、12和24 h处死大鼠进行肺泡灌洗,采用酶联免疫吸附法测定支气管肺泡灌洗液(BALF)中凝血酶-抗凝血酶复合物(TAT)、组织型纤溶酶原激活物(t-PA)、尿激酶型纤溶酶原激活物(u-PA)、纤溶酶原激活物抑制剂-1(PAI-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平.结果 损伤后6 h,模型组BALF中TAT浓度(μg/L:3.346±0.585)最高,其次是HT组(2.764±0.100),HP组(2.564±0.216)最低(均P＜0.05);HP组t-PA(μg/L:3.037±0.524)最高,HT组(2.494±0.191)其次,模型组(1.716±0.125)最低(均P＜0.05);HP组u-PA(μg/L)高于模型组(0.411±0.118比0.303±0.049,P＜0.05);HP组PAI-1(μg/L)明显低于HT组和模型组(2.296±0.246比2.597±0.425、2.834±0.198,均P＜0.05),直至12 h时HP组仍低于HT组(1.273±0.441比1.817±0.252,P＜0.05);HT组和HP组TNF-α(ng/L)显著低于模型组(68.154±3.915、36.990±6.539比77.001±4.485),且HP组低于HT组(均P＜0.05),至12 h HP组(15.287±4.754)仍最低,与HT组和模型组(26.756±5.336、23.674±4.398)比较差异有统计学意义(均P＜0.05).模型组、HT组和HP组各时间点IL-6水平均无明显差异.结论 内毒素性ALI大鼠雾化吸入UFH后起到了抑制局部凝血、减弱纤溶抑制、促进纤溶、降低炎症反应的作用,预防性吸人UFH比治疗性吸入效果更显著,最佳效应时间在注射后6 h.

Abstract：

Objective To observe the local changes in alveoli in intravenous endotoxin-induced acute lung injury (ALI) rat model after inhalation of aerosolized unfractioned heparin(UFH), and to observe its effects on coagulability, fibrinolysis and inflammatory response. Methods Eighty-seven male Wistar rats were divided into groups according to table of random number: sham, model, heparin therapy (HT) and heparin prophylaxis(HP). Endotoxin-induced ALI model was reproduced by intravenous administration of lipopolysaccharide (LPS). Rats in Hp and HT groups received aerosolized UFH before and after injection with LPS respectively, while rats in both sham and model groups inhaled aerosolized normal saline. Rats in each group were respectively sacrificed at 6, 12 and 24 hours after intravenous administration of LPS, and bronchoalveolar lavage fluid (BALF) was collected. Enzyme-linked immunosorbent assay was used to measure the level of thrombin-antithrombin (TAT), tissue-type plasminogen activator (t-PA),urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor- (TNF-α), interleukin-6 (IL-6) in BALF. Results At.6 hours after injury, the level of TAT (μg/L) in model group (3.346±0. 585) was highest, that in HT group (22. 764±0. 100) was higher, and that in HP group (2. 564±0. 2216) was lowest in BALF (all P＜0. 05). The t-PA(μg/L) concentration in HP group (3.037±0. 5224) was highest, that in HT group (22. 494±0. 191) was higher, and that in model group (1. 716±0. 1225) was lowest (all P＜0. 05). Compared with model group, u-PA (μg/L) level in HP group dramatically enhanced (0. 411±0. 118 vs. 0. 303±0. 049, P＜0. 05). The concentration of PAI-1 (μg/L) in HP group was significantly lower than that of HT and model groups (22. 2296 ± 0. 2246 vs. 22.597±0. 4225,2.834±0.198, both P＜0. 05). In HP group, it was still lower than that in HT group at 12 hours (1.2273±0. 441 vs. 1. 817±0. 252, P＜0. 05). TNF-α(ng/L) levels in HT and HP groups were markedly lower compared with model group (68. 154±3. 915, 36. 990±6. 539 vs. 77. 001±4. 485) at 6 hours, and the level in HP group was lower than that in HT group (all P＜0. 05). TNF-α concentration in HP group was still the lowest at 122 hours (15.2287±4. 754), and there was significant difference compared with HT and model groups (26. 756±5. 336, 23. 674±4. 398, both P＜0. 05). The levels of IL-6 were not distinctively different among model, HT and HP groups at various time-points. Conclusion It was proved that inhalation of aerosolized UFH resulted in lowering local coagulability, alleviating fibrinolytic depression, improving fibrinolysis, and attenuating inflammation in endotoxin-induced ALI rat model. More prominent results will be obtained when it was use as a prophylactic measure. The optimal time of usage is 6 hours after endotoxin injection.     

高浓度氧对未成年大鼠肺部炎症反应的影响

目的 探讨高浓度氧对未成年大鼠肺部炎症反应的影响.方法 将40只出生21 d的SD大鼠按随机数字表法分为空气对照组及高氧暴露12、24、48、72 h组,每组8只,分别将大鼠置于空气和常压高氧箱(氧含量达92%～94%)中.于相应时间点采用放血法处死大鼠后取肺组织,并行支气管肺泡灌洗.采用硫代巴比妥酸法和比色法分别测定肺组织丙二醛(MDA)含量及髓过氧化物酶(MPO)活性;采用酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和IL-10含量;观察肺组织病理改变,并进行肺损伤评分.结果 与空气对照组比较,高氧暴露12 h肺组织MDA含量(mmol/g)即显著升高(2.24±0.43比1.57±0.31),MPO活性(U/g)于高氧暴露24 h显著升高(1.24±0.25比0.69±0.22),并均随高氧暴露时间延长逐渐增加(P<0.05或P<0.01).BALF中TNF-α、IL-6和IL-10含量于高氧暴露24 h时较空气对照组显著增加[TNF-α(ng/L):135.2±44.0比94.5±22.3,IL-6(ng/L):73.1±14.2比55.7±17.3,IL-10(ng/L):67.9±21.7比48.2±7.6,P<0.05或P<0.01];但高氧暴露48 h时较24 h时显著降低(48 h时BALF中TNF-α、IL-6、IL-10分别为105.4±17.0,54.3±17.4,50.9±6.9,均P<0.05).高氧暴露12 h时肺损伤评分(分)即较空气对照组显著升高(4.5±1.4比1.3±0.5),并随高氧暴露时间延长进一步升高(P<0.05或P<0.01).结论 高浓度氧可引起未成年大鼠肺部炎症损伤;炎症细胞因子的出现高峰均在高氧暴露24 h.     

烟碱对心肌缺血/再灌注损伤大鼠炎症细胞因子的影响

目的 探讨烟碱对心肌缺血/再灌注(I/R)损伤大鼠炎症细胞因子的影响.方法 50只健康雄性SD大鼠按随机数字表法分为假手术组、I/R组、烟碱高剂量(400μg/kg)组、烟碱低剂量(40μg/kg)组及α-银环蛇毒素(α-BGT,1μg/kg)组5组,每组10只.采用结扎心脏左冠状动脉前降支30 min、再灌注90 min制作大鼠心肌I/R损伤模型;假手术组仅穿线不结扎.制模前30 min各药物组颈静脉注射相应剂量药物干预,假手术组和I/R组给予等量生理盐水.于再灌注末取右颈动脉血,测定肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、IL-10浓度和肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)活性;然后处死动物,取缺血区心肌组织测定髓过氧化物酶(MPO)活性;采用免疫组化和逆转录-聚合酶链反应检测心肌组织细胞间黏附分子-1(ICAM-1)蛋白及mRNA表达,并观察心肌超微结构.结果 与假手术组比较,I/R组血浆TNF-α、IL-8、IL-10、CK-MB、cTnI、心肌MPO活性及ICAM-1蛋白和mRNA表达均显著升高[TNF-α(ng/L):158.7±32.7比31.5±5.8,IL-8(ng/L):0.71±0.06比0.30±0.04,IL-10(ng/L):69.0±7.8比41.4±4.3,CK-MB(U/L):2 540±169比1 120±102,cTnI(μg/L):26.2±4.6比0.9±0.2,MPO(U/g):4.2±0.6比1.6±0.4,ICAM-1蛋白:0.210±0.025比0.100±0.018,ICAM-1 mRNA:1.82±0.23比1.18±0.20,P＜0.05或P＜0.01],病理学显示心肌组织损伤较重.与I/R组比较,烟碱高剂量组血浆TNF-α、IL-8降低[TNF-α(67.3±9.8)ng/L,IL-8(0.47±0.04)ng/L],IL-10升高[(147.5±12.5)ng/L],CK-MB、cTnI及心肌MPO活性、ICAM-1蛋白和mRNA均降低[CK-MB(1 282±145)U/L,cTnI(4.7±1.4)μg/L,MPO(2.5±0.4)U/g,ICAM-1蛋白0.140±0.026,ICAM-1 mRNA 1.31±0.25,P＜0.05或P＜0.01],心肌组织损伤减轻;而烟碱低剂量组和α-BGT组上述指标与I/R组比较差异无统计学意义.结论 烟碱可阻断内皮细胞表达黏附分子,阻断中性粒细胞黏附、游出,改善抗炎/促炎反应平衡,从而拮抗大鼠心肌I/R损伤时的过度炎症反应.     

天冬氨酸特异性半胱氨酸蛋白酶3抑制剂对腹腔感染脓毒症急性肾损伤小鼠血清炎症细胞因子的影响

目的 观察天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)抑制剂Ac-DEVD-CHO对腹腔感染脓毒症急性肾损伤(AKI)小鼠血清炎症细胞因子的影响. 方法 将102只C57BL/6小鼠按随机数字表法均分成假手术组、模型组和caspase-3抑制剂组(CI组)3组.采用盲肠结扎穿孔术(CLP)制备脓毒症AKI小鼠模型;CI组在CLP术前给予Ac-DEVD-CHO 4 μg/g腹部皮下注射进行干预.各组均于术后6、12、24 h检测血尿素氮(BUN)、肌酐(Cr)浓度;用酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-6、IL-10)浓度;用流式细胞仪检测肾组织细胞凋亡率;用实时荧光定量聚合酶链反应(PCR)检测肾组织caspase-3 mRNA表达;并观察小鼠4 d和7 d的存活率. 结果 与假手术组比较,模型组各时间点血清BUN、TNF-α、IL-6、IL-10及肾组织细胞凋亡率、caspase-3 mRNA表达均显著升高,术后6 h Cr显著升高,4 d、7 d存活率显著下降.与模型组比较,CI组各时间点BUN显著下降;术后6 h Cr显著下降,12 h和24 h恢复至假手术组水平;各时间点血清TNF-α、IL-6水平降低,IL-10水平升高[TNF-α(μg/L)6 h:436.2±64.2比653.6±8.9,12 h:233.4±85.4比579.7±137.1,24 h:151.0±90.3比551.0±119.8;IL-6(μg/L) 6 h:1 033.2±345.8比1 595.3±159.4,12 h:366.3±68.3比1 330.7±249.8,24 h:241.2±208.4比815.3±572.7;IL-10(μg/L)6 h:33.6±10.4比26.6±4.5,12 h:37.2±5.0比24.5±4.3,24 h:38.3±5.5比18.2±1.6,均P<0.05];肾组织细胞凋亡率及caspase-3 mRNA表达显著下降[凋亡率6 h:(13.9±3.2)%比(18.3±1.4)%,12 h:(10.5±3.6)%比(15.9±3.5)%,24 h:(8.4±1.8)%比(12.5±2.1)%;caspase-3 mRNA 6 h:1.95±0.16比3.84±0.35,12 h:1.89±0.19比3.97±0.73,24 h:2.01±0.20比4.97±0.24,均P<0.05],并提高了小鼠4 d存活率(80%比20%),7 d存活率无变化(20%比20%). 结论 caspase-3抑制剂对腹腔感染脓毒症AKI小鼠血清促炎/抗炎细胞因子水平的影响可能与降低肾组织细胞凋亡有关.     

危重病患者血中生长抑素水平检测的临床意义

目的 探讨危重病患者血中生长抑素(SST)水平与机体炎症反应和病情严重度的关系以及对预后的评估价值.方法 选择60例急性生理学与慢性健康状况评分系统Ⅱ(APACHE Ⅱ)评分≥8分的危重病患者,按照APACHE Ⅱ评分分为3组:轻度组(<16分)23例,中度组(16～20分)20例,重度组(>20分)17例;按预后分为死亡组(13例)与存活组(47例).另选择20例健康自愿者作为对照.用放射免疫法检测血中SST水平;用酶联免疫吸附法(ELISA)检测血中肿瘤坏死因子-α(TNF-α)、白细胞介索-6(IL-6)水平.结果 危重病组患者血中SST水平明显低于健康对照组[(18.2±17.6)ng/L比(224.8±130.2)ng/L,P<0.053;TNF-α、IL-6水平明显高于健康对照组[TNF-α:(32.4±14.2)ng/L比(14.2±5.7)ng/L,IL-6:(131.6±42.7)ng/L比(65.8±24.3)ng/L,P均<0.053;SST与TNF-α、IL-6水平均呈显著负相关(r1=-0.682,r2=-0.894,P均<0.01).重度组血中SST水平明显低于轻、中度组C(8.1±7.2)ng/L比(24.7±15.9)ng/L、(19.2±22.1)ng/L,P均<0.053;TNF-α、IL-6水平明显高于轻度组[TNF-α:(39.0±16.4)ng/L比(28.9±10.9)ng/L,IL-6:(156.05=49.6)ng/L比(111.5±32.6)ng/L,P均<0.053;SST水平与APACHE Ⅱ评分呈显著负相关(r=-0.327,P<0.05).死亡组血中SST水平明显低于存活组[(6.4±5.5)ng/L比(21.5±18.4)ng/L,P<0.053;TNF-α、lL-6水平虽高于存活组,但差异均无统计学意义.结论 危重病患者血中SST水平可以反映病情严重程度,对评估患者预后也具有临床价值.     

胍丁胺对酵母多糖诱导急性肺损伤的器官保护作用

目的 探讨胍丁胺(AGM)对酵母多糖(ZYM)诱导小鼠全身炎症反应和急性肺损伤(ALI)时器官的保护作用.方法 将32只成年雄性C57BL/6小鼠按随机数字表法分为正常对照组[磷酸盐缓冲液(PBS)0.5 ml]、AGM对照组(AGM 200 mg/kg)、模型组(ZYM 500 mg/kg+ PBS 0.5 ml)、AGM治疗组(ZYM 500 mg/kg+AGM 200 mg/kg)4组,每组8只.AGM、ZYM、PBS为腹腔注射.于给药12h后采用酶联免疫吸附法(ELISA)测定血清和腹腔渗出液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及一氧化氮(NO)含量;同时测定肺组织中TNF-α、IL-6、髓过氧化物酶(MPO)活性和核转录因子-kB p65(NF-kB p65)的DNA结合活性,并观察肺组织病理改变.结果 ZYM注射后12 h小鼠精神萎靡、活动减少、饮水减少;而AGM治疗组小鼠精神状况、活动、饮水均好转.AGM治疗能降低ZYM诱导的血清及腹腔渗出液的TNF-α(ng/L:252.6±32.1比421.7±76.7,295.7±78.6比592.0±84.3,均P＜0.05)、IL-6(ng/L:2 198.8±281.8比4 725.3±615.4,19 829.3±3 647.0比47 751.3±5 264.8,均P＜0.05)和NO (μmol/L:33.2±4.3比50.2±5.2,14.0±3.6比45.4±5.2,均P＜0.05)水平升高;AGM治疗也能够抑制ZYM引起的肺组织TNF-α(ng/L:245.7±39.1比378.3±67.6,P＜0.05)、IL-6 (ng/L:810.3±175.6比1 172.4±203.3,P＜0.05)、MPO活性(ng/mg:24.9±4.4比37.3±5.8,P＜0.05)和NF-kB p65(吸光度值:0.272±0.029比0.347±0.037,P＜0.05)升高.正常对照组和AGM对照组间各指标无明显差异.ZYM可导致肺组织出现严重的炎症改变,包括血管扩张、中性粒细胞浸润;AGM治疗后肺组织损伤明显减轻.结论 AGM能够缓解ZYM诱导的小鼠全身炎症反应和ALI的严重程度.     

还原型谷胱甘肽对脓毒症大鼠肺部超微结构及血中细胞因子水平的影响

目的 探讨还原型谷胱甘肽(GSH)对脓毒症大鼠急性肺损伤保护作用的机制.方法 采用盲肠结扎穿孔术(CLP)复制SD大鼠脓毒症模型.按随机数字表法将大鼠分为假手术组、模型组、GSH组、左氧氟沙星(LEV)组;分别于术后3、6、12、24 h各取7只大鼠心脏血检测血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平,电镜下观察术后24 h大鼠肺组织超微结构的改变.结果 与假手术组C(132±9)μg/L]比较,模型组术后6 h血浆TNF-α水平((227±28)μg/L]显著升高(zp<0.01);GSH治疗组((144±28)μg/L]较模型组显著降低,且明显低于LEV组((214±48)μg/L,均P<0.01];各组间术后3、12、24 h TNF-α水平比较均无明显差异.与假手术组((135.43±40.08)μg/L]比较,模型组术后3 h血浆IL-6水平((267.65±72.87)μg/L]显著升高(P<0.01);GSH治疗组[(191.97±62.98)μg/L]较模型组显著降低,且明显低于LEV组[(268.75±74.67)μg/L,均P<0.05];各组间术后6、12、24 h IL-6水平均无明显差异.模型组大鼠肺组织超微结构发生显著变化,尤其是细胞内线粒体出现水肿甚至空泡变性;GSH组大鼠肺组织超微结构的改变轻微.结论 TNF-α和IL-6在脓毒症大鼠肺损伤发生机制中起重要作用,GSH有明显的治疗效果.