杀菌/通透性增加蛋白对休克大鼠肺组织细胞因子mRNA表达的影响

作者在大鼠失血性休克（４ｋＰａ，１８０分钟）模型上，观察了重组杀菌／通透性增加蛋白（ＢＰＩ）对肺组织肿瘤坏死因子（ＴＮＦ）、白介素－６（ＩＬ－６）ｍＲＮＡ表达及急性肺损伤的影响，并对肠源性内毒素血症与炎症细胞因子诱发的关系进行了探讨。结果显示：失血性休克可导致血浆内毒素含量显著升高，肺组织ＴＮＦ、ＩＬ－６ｍＲＮＡ表达分别在复苏后２、８小时明显增多（Ｐ＜０．０５～０．０１）；给予ＢＰＩ治疗则完全中和休克所致内毒素血症，并不同程度地抑制肺组织ＴＮＦ、ＩＬ－６ｍＲＮＡ的表达（Ｐ＜０．０５～０．０１）；肺毛细血管通透性与髓过氧化物酶活性均明显降低。作者认为，ＢＰＩ可有效防止失血性休克诱发的急性肺损伤，其作用机制可能与抑制肠源性内毒素血症所介导的局部组织炎症细胞因子基因表达有关。     

杀菌/通透性增加蛋白对内毒素休克大鼠微循环灌注的保护作用

目的 探讨杀菌／通透性增加蛋白（ＢＰＩ）对内毒素休克时血液动力学和内脏微徨灌注的保护作用及其机制。方法 采用大鼠内毒素休克模型,动态观察血液动力学、内脏微循环灌注量和血浆生物喋吟、一氧化氮（ＮＯ）水平的变化。结果 休克早期给予重组ＢＰＩ,可显著提高平均支流压、心脏指数及每搏输出量,明显改善肝、肾、小肠抽徨灌注量及动物预后（Ｐ〈０．０５～０．０１）,循环ＮＯ产生亦不同程度受到抑制。结论 早期应用ＢＰ     

杀菌性通透性增强蛋白对内毒素休克大鼠重要器官的保护作用

目的 探讨杀菌性通透性增强蛋白（ＢＰＩ）对内毒素休克大鼠重要器官功能的保护作用,为ＢＰＩ的临床研究提供实验依据。方法 Ｗｉｓｔａｒ大鼠２０只,１ 次性动脉内注射大鼠肝菌内毒素（ＬＰＳ）１２．５ｍｇ／ｋｇ,复制内毒素休克模型。注射ＬＰＳ后即刻,１例性动脉内注射ＢＰＩ５ｍｇ／ｋｇ或等容量生理盐水。结果 （１）ＢＰＩ治疗组动物存活时间延长,动物２４小时存活率为８０％,显著高于生理盐水对照组的２０％;（２     

不同剂量6%HES 130/0.4容量治疗对失血性休克大鼠肺损伤的影响

目的 评价不同剂量6%羟乙基淀粉130/0.4(6%HES 130/0.4)容量治疗对失血性休克大鼠肺损伤的影响.方法 健康雄性SD大鼠24只,随机分为4组(n=6),假手术组(S组)、乳酸钠林格氏液组(RS组)和6%HES130/0.4 33ml/kg组(H1组)、6%HES130/0.4 50ml,kg组(H2组).除S组外,RS组、H1组和H2组均经右颈总动脉放血,制备失血性休克模型.于模型制备成功后RS组静脉输注3倍最大放血量的乳酸钠林格氏液;H1组和H2组分别静脉输注33、50ml/kg 6%HES 130/0.4和乳酸钠林格氏液(总量均为3倍最大放血量),容量治疗时间45 min.于放血前(T0,基础状态)、容量治疗结束后2 h(T1)、3 h(T2)时采集动脉血样,进行血气分析,计算PaO2/FiO2;最后一次采集血样后,进行支气管肺泡灌洗,测定支气管肺泡灌洗液(BALF)蛋白浓度,取肺组织测定湿,干重比(W/D比值)、TNF-α、m-1β和IL-10的含量,光镜下观察肺组织病理学结果.结果 与S组比较,RS组、H1组和H2组肺组织TNF-α、IL-1β、IL-10含量、BALF蛋白浓度和W/D比值升高,RS组T1.2时PaO2/FiO2降低,H2组T2时PaO2/FiO2降低(P<0.05),H1组T1.2时PaO2/FiO2差异无统计学意义(P>0.05);与RS组比较,H1组和H2组肺组织TNF-α、IL-1β含量、BALF蛋白浓度和W/D比值降低,H1组T1.2时,H2组T1时PaO2/FiO2升高(P<0.05).与H1组比较,H2组T2时PaO2/FiO2降低,肺组织IL-10含量降低(P<0.05).H1组和H2组肺组织病理损伤程度轻于RS组,其中H1组病理学损伤程度最轻.结论 6%HES 130/0.4 33和50 ml/kg容量治疗均可减轻失血性休克大鼠肺损伤,33 ml/kg效果更好.     

盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时TLR4 mRNA表达的影响

目的 探讨盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时Toll样受体4(TLR4)mRNA表达的影响.方法 健康SD大鼠40只,体重200～250 g,随机分为5组(n=8):假手术组(S组)、失血性休克致急性肺损伤组(ALI组)和低、中、高剂量盐酸戊乙奎醚预先给药组(P1～3组).S组仅行动静脉穿刺,不放血,ALI组股动脉放血至35～45 mm Hg制备急性肺损伤模型,P1～3组分别于放血前30 min股静脉注射盐酸戊乙奎醚0.3、1.0、3.0 mg/kg,随后制备急性肺损伤模型.各组复苏后4 h时处死大鼠取肺,称重后计算肺湿干重比,检测TLR4 mRNA和NF-κB p65蛋白的表达水平,观察病理学结果.结果 与S组比较,ALI组和P1组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比升高(P＜0.05或0.01),P2.3组差异无统计学意义(P＞0.05);与ALI组比较,P2,3组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比降低(P＜0.05或0.01);P2组和P3组上述指标比较差异无统计学意义(P＞0.05).P2,3组肺组织病理学损伤程度较ALI组明显减轻.结论 盐酸戊乙奎醚预先给药可通过抑制肺组织TLR4 mRNA表达上调,进而降低NF-κB活性,从而减轻失血性休克诱发大鼠的急性肺损伤.     

不同剂量6%HES 130/0.4容量治疗对失血性休克大鼠肺损伤的影响

Objective To evaluate the effects of volume therapy with different doses of 6% hydroxyethyl starch 130/0.4 (6% HES 130/0.4) on lung injury in a rat model of hemonhagic shock.Methods Twenty-four male SD rats weighing 220-300 g were randomly divided into 4 groups ( n = 6 each) : group I sham operation (group S); group II Ringer's solution (group RS); group HI and IV 2 HES groups (group H1, H2 ). The animals were anesthetized with intraperitoneal 1% sodium pentobarbital 45 ing/kg. Right common carotid artery (CCA) and left femoral vein were cannulated for blood letting, MAP monitoring, fluid administration and blood sampling. Hemonhagic shock was induced by withdrawing blood from right CCA in group II , III and IV . MAP was reduced to 35-45 mmHg which was maintained for 90 min. In group RS, hemorrhagic shock was resuscitated with Ringer's solution 3 times of the volume of blood withdrawn, while group H1 and H2 received HES 33 and 50 ml/kg respectively and Ringer' s solution (the total volume was equal to 3 times of the volume of blood removed) . Arterial blood samples were taken before blood letting (T0 , baseline), and at 2, 3 h after volume therapy (T1,2) for blood gas analysis and PaO2/FiO2 was calculated. The animals were then sacrificed by exsanguination and the lungs were immediately removed for microscopic examination and determination of protein concentration in broncho-alveolar lavage fuid (BALF), W/D lung weight ratio and TNF-α, IL-1 β and IL-10 contents in the lung.Results TNF-α, IL-1β and IL-10 content in the lung, protein concentration in BALF and W/D ratio were significantly higher in group RS, H1 and H2, while PaO2/FiO2 was significantly lower at T,2 in group RS and at T2 in group H2 than in group S (P < 0.05). TNF-α and IL-1β contents in the lung, protein concentration in BALF and W/D ratio were significantly lower in group H1 and H2 , while PaO2/FiO2 was significantly higher at T,i2 in group H1 and at T1 in group H2 than in group RS (P <0.05) . PaO2/FiO2 at T2 and IL-10 content in the lung were significantly lower in group H2 than in group H, ( P < 0.05) . The lung damage was significantly ameliorated in group H1 and H2 especially in group H, as compared with group RS. Conclusion Volume therapy with 6% HES 130/0.4 33 or 50 ml/kg can attenuate lung injury in a rat model of hemorrhagic shock and the efficacy of 33 ml/kg is better.     

不同剂量6%HES 130/0.4容量治疗对失血性休克大鼠肺损伤的影响

Objective To evaluate the effects of volume therapy with different doses of 6% hydroxyethyl starch 130/0.4 (6% HES 130/0.4) on lung injury in a rat model of hemonhagic shock.Methods Twenty-four male SD rats weighing 220-300 g were randomly divided into 4 groups ( n = 6 each) : group I sham operation (group S); group II Ringer's solution (group RS); group HI and IV 2 HES groups (group H1, H2 ). The animals were anesthetized with intraperitoneal 1% sodium pentobarbital 45 ing/kg. Right common carotid artery (CCA) and left femoral vein were cannulated for blood letting, MAP monitoring, fluid administration and blood sampling. Hemonhagic shock was induced by withdrawing blood from right CCA in group II , III and IV . MAP was reduced to 35-45 mmHg which was maintained for 90 min. In group RS, hemorrhagic shock was resuscitated with Ringer's solution 3 times of the volume of blood withdrawn, while group H1 and H2 received HES 33 and 50 ml/kg respectively and Ringer' s solution (the total volume was equal to 3 times of the volume of blood removed) . Arterial blood samples were taken before blood letting (T0 , baseline), and at 2, 3 h after volume therapy (T1,2) for blood gas analysis and PaO2/FiO2 was calculated. The animals were then sacrificed by exsanguination and the lungs were immediately removed for microscopic examination and determination of protein concentration in broncho-alveolar lavage fuid (BALF), W/D lung weight ratio and TNF-α, IL-1 β and IL-10 contents in the lung.Results TNF-α, IL-1β and IL-10 content in the lung, protein concentration in BALF and W/D ratio were significantly higher in group RS, H1 and H2, while PaO2/FiO2 was significantly lower at T,2 in group RS and at T2 in group H2 than in group S (P < 0.05). TNF-α and IL-1β contents in the lung, protein concentration in BALF and W/D ratio were significantly lower in group H1 and H2 , while PaO2/FiO2 was significantly higher at T,i2 in group H1 and at T1 in group H2 than in group RS (P <0.05) . PaO2/FiO2 at T2 and IL-10 content in the lung were significantly lower in group H2 than in group H, ( P < 0.05) . The lung damage was significantly ameliorated in group H1 and H2 especially in group H, as compared with group RS. Conclusion Volume therapy with 6% HES 130/0.4 33 or 50 ml/kg can attenuate lung injury in a rat model of hemorrhagic shock and the efficacy of 33 ml/kg is better.     

不同液体复苏对失血性休克-内毒素二次打击大鼠急性肺损伤的影响

目的 比较不同液体复苏对失血性休克-内毒素二次打击大鼠急性肺损伤的影响.方法 60只雄性SD大鼠,体重250～280 g,随机分为5组(n=12):假手术组(S组)、失血性休克-内毒素组(SL组)、乳酸钠林格氏液组(LR组)、7.5%氯化钠组(HS组)和羟乙基淀粉(HES)130/0.4组(HES组).分为院前期(90 min)、院内复苏期(1 h)和复苏后观察期(3.5 h).院前期:SL组、LR组、HS组和HES组颈总动脉放血建立失血性休克模型(维持MAP 35～45 mm Hg 60 min)后,气管内注射内毒素2mg/kg,同时断尾,注射内毒素后即刻分别经30 min静脉输注3倍放血量的乳酸钠林格氏液、7.5%氯化钠4ml/kg和等放血量的6%HES 130/0.4;院内复苏期:结扎尾部断端止血,在1 h内回输全部放出的血液及等放血量的0.9%氯化钠;复苏后观察期3.5 h时采集动脉血样,进行血气分析,计算肺组织湿/干重量比(W/D)和肺通透指数(PPI),测定肺泡灌洗液(BALF)蛋白浓度和肺组织AQP-1 mRNA和AQP-5mRNA表达水平,光镜下观察肺组织病理学结果,记录大鼠存活情况.结果 与SL组和LR组比较,HS组和HES组MAP、pH值、PaO2和SaO2升高,血乳酸浓度、BE、W/D、PPI和BALF蛋白浓度降低,HS组肺组织AQP-1 mRNA表达上调,HES组肺组织AQP-1 mRNA和AQP-5 mRNA表达上调,大鼠存活率升高(P＜0.05或0.01);与HS组比较,HES组W/D降低,AQP-5 mRNA表达上调,大鼠存活率升高(P＜0.05),肺组织损伤程度减轻.结论 6%HES 130/0.4和7.5%氯化钠可减轻失血性休克-内毒素二次打击大鼠急性肺损伤,6%HES 130/0.4的效果更好,其机制与抑制AQP-1和(或)AQP-5表达下调有关;而乳酸钠林格氏液对其无效.     

利多卡因对失血性休克大鼠肺损伤的保护作用

目的 探讨利多卡因对失血性休克大鼠肺损伤的保护作用。方法 80只雄性Wistar大鼠建立失血性休克模型后,随机分为四组,假手术组(Ⅰ组,n=8)、休克组(Ⅱ组,n=8)、生理盐水组(Ⅲ组,n=32)、利多卡因组(Ⅳ组,n=32)。Ⅰ组于假手术后,Ⅱ组于休克60 min,Ⅲ、Ⅳ组分别于复苏开始后2、4、8、12 h,测定中性粒细胞(PMNs)表面粘附分子CD11b/CD18表达、肺组织中髓过氧化物酶(MPO)活性,并采用光镜和透射电镜观察肺组织的病理学改变。结果 与Ⅰ组比较,Ⅲ组、Ⅳ组复苏后各时点PMNs表面CD11b/CD18表达均升高(P<0.01),Ⅱ组差异无显著性(P>0.01),Ⅱ组、Ⅲ组、Ⅳ组复苏后各时点肺组织中MPO活性升高(P<0.05或0.01)。与Ⅲ组比较,Ⅳ组复苏后同一时点、Ⅰ组、Ⅱ组PMNs表面CD11b/CD18表达及肺组织中MPO活性降低(P<0.01)。结论 小剂量利多卡因可以抑制失血性休克大鼠PMNs表面CD11b/CD18的表达,减少PMNs在肺组织中的浸润,从而减轻肺损伤。     

重组杀菌／通透性增加蛋白对内毒素休克中肝脏一氧化氮合酶的 …

目的：探讨重组杀菌／通透性增加蛋白（ｒＢＰＩ２１）对内毒素休克中肝组织一氧化氮合酶（ＮＯＳ）的影响及其意义。方法 大鼠腹腔注射大肠杆菌内毒素（１５．０ｍｇ／ｋｇ）复制内毒素休克模型，动物随机分成正常对照组、内毒素休克组和ｒＢＰＩ２１治疗组。检测肝组织ＮＯＳ活性、三磷酸鸟苷环水解酶Ｉ（ＧＴＰ－ＣＨＩ）活性及生物喋呤含量，同时还观察肝脏微循环血流灌注量的改变。结果 内毒素攻击后肝组织诱生型ＮＯＳ（ｉＮ     

Treatment with recombinant bactericidal/permeability-increasing protein to prevent endotoxin-induced mortality in bile duct-ligated rats.

BACKGROUND: Operation in patients with obstructive jaundice is associated with substantial morbidity because of increased susceptibility to endotoxin (lipopolysaccharide) and the inflammatory cascade. Different interventions to reduce endotoxemia and cytokine induction, and resulting complications, have been studied. Bactericidal/permeability-increasing protein (BPI) is a naturally occurring endotoxin-binding protein produced in neutrophils. It binds endotoxin, neutralizing the activity and inhibiting cytokine production by mononuclear cells. In experimental endotoxemia in animals and in healthy human volunteers, BPI has shown a protective effect. The aim of this study was to determine whether BPI could protect against increased endotoxin sensitivity in rats with obstructive jaundice and reduce endotoxin-induced mortality. STUDY DESIGN: Male Wistar rats were used. Intraperitoneal Escherichia coli 2mg/kg was given 1 week after sham operation or bile duct ligation (BDL). Three groups were studied: sham, BDL with placebo, and BDL with 5 mg/kg recombinant BPI21. RESULTS: BDL rats were jaundiced (mean bilirubin 186 micromol/L; no difference between BDL rats without or with BPI). Bilirubin remained less than 1 micromol/L in sham-operated rats (p = 0.002). Endotoxin levels were 3.4pg/mL in sham controls and 3.1 pg/mL in BDL rats before administration of lipopolysaccharide (p = NS). Two hours after administration, levels were 615.4ng/mL in placebo BDL rats and 10 times less in BPI-treated BDL rats, at 60.2ng/mL (p=0.03). The same trend was found at 6 hours. At 24 hours, mortality was 1 of 6 in sham-operated rats (15%) versus 8 of 11 in untreated BDL rats (75%). BPI intervention reduced the death rate to 1 of 12 BDL rats (8%) (p = 0.003). CONCLUSIONS: Intraperitoneal recombinant BPI21 in rats having BDL reduced endotoxin-induced mortality from 75% to 8%, a death rate comparable to that in nonjaundiced rats. BPI could be an interesting perioperative treatment in clinical obstructive jaundice.     

Pathogenesis of hemorrhage-induced bacteria/endotoxin translocation in rats. Effects of recombinant bactericidal/permeability-increasing protein.

OBJECTIVES: This study was conducted to determine the role of gut-derived bacteria/endotoxin in the pathogenesis of the multiple-organ damage and mortality, the possible beneficial effect of recombinant bactericidal/permeability-increasing protein (rBPl21), and whether neutralizing endotoxemia by rBPl21 treatment influences tumor necrosis factor (TNF) formation in rats after hemorrhagic shock and resuscitation. SUMMARY BACKGROUND DATA: Hypovolemic shock might be associated with bacterial or endotoxin translocation as well as systemic sepsis. Similar to bactericidal/permeability-increasing (BPl) protein, rBPl21 has been found to bind endotoxin and inhibit TNF production. METHODS: A rat model of prolonged hemorrhagic shock (30 to 35 mm Hg for 180 min) followed by adequate resuscitation was employed. Recombinant bactericidal/permeability-increasing protein was administered at 5 mg/kg intravenously. The control group was treated similarly to the BPl group, but received thaumatin as a protein-control preparation in the same dose as rBPl21. RESULTS: Immediately after resuscitation (230 min), plasma endotoxin levels in the control group (61.0 +/- 16.3 pg/mL) were almost neutralized by rBPl21 treatment (13.8 +/- 4.8 pg/mL, p < 0.05). Plasma TNF levels were not significantly influenced by rBPl21 treatment. The 48-hour survival rate was 68.8% in the treatment group versus 37.5% in the control group (p = 0.08). Microscopic histopathologic examination revealed relatively minor damage to various organs in the treatment group. CONCLUSIONS: These data suggest that hemorrhagic shock may lead to bacterial/endotoxin translocation with concomitant TNF formation, endogenous endotoxemia may play an important role in the pathogenesis of multiple-organ failure after shock and trauma, TNF formation at an early stage might be related mainly to mechanisms other than Kupffer's cells activation via lipopolysaccharide, and rBPl21 might be a useful therapeutic agent against endogenous bacteria/endotoxin related disorders in severe hemorrhagic shock.     

Natural antibiotics and insulin sensitivity: the role of bactericidal/permeability-increasing protein

The innate immune system can immediately respond to microorganism intrusion by helping to prevent further invasion. Bactericidal/permeability-increasing protein (BPI) is a major constituent of neutrophils that possesses anti-inflammatory properties. Inflammation is increasingly recognized as a component of the metabolic syndrome. We hypothesized that the production of BPI could be linked to insulin sensitivity and glucose tolerance. We studied circulating BPI across categories of glucose tolerance. We also studied whether these cross-sectional associations were of functional importance. For this reason, we investigated circulating bioactive lipopolysaccharide and the effects of changing insulin action-after treatment with an insulin sensitizer (metformin)-on circulating BPI in subjects with glucose intolerance. Finally, we tested whether a 3'-untranslated region (UTR) BPI polymorphism led to differences in BPI and insulin action among nondiabetic subjects. Age- and BMI-adjusted circulating BPI was significantly lower among patients with type 2 diabetes. Circulating BPI correlated negatively with fasting and postload glucose and insulin concentrations. In subjects with glucose intolerance, BPI was also linked to BMI, waist-to-hip ratio, and age- and BMI-adjusted insulin sensitivity. Bioactive lipopolysaccharide was negatively correlated with circulating BPI (r = -0.57, P < 0.0001) and positively with plasma lipopolysaccharide-binding protein (r = 0.54, P = 0.002). In parallel to improved insulin sensitivity, plasma BPI significantly increased in the metformin group but not in the placebo group. A 3'-UTR BPI polymorphism was simultaneously associated with plasma BPI concentration, waist-to-hip ratio, fasting and postload insulin concentration, fasting plasma triglycerides, and insulin sensitivity. These findings suggest that this component of the innate immune system is associated with metabolic pathways.     

Structure/function studies of an endotoxin-neutralizing peptide derived from bactericidal/permeability-increasing protein

BACKGROUND: Bactericidal/permeability-increasing protein, BPI, has a beta-turn with alternating cationic and hydrophobic residues in its lipopolysaccharide (endotoxin, LPS)-binding domain. A peptide, betapep25, was designed with 9 residues of the LPS-binding domain of BPI flanked by beta-turn-inducing elements. Thereafter, we sought to use single amino acid substitutions to identify residues that are important for the biological activities of betapep25. METHODS: Single alanine or norleucine replacement "walkthrough" peptides based on betapep25 were generated and tested for their ability to kill P aeruginosa and to neutralize endotoxin. RESULTS: Substitution of all lysines inhibited bactericidal activity. Inhibition of LPS-neutralizing activity was seen in 9 peptides in which an alanine or norleucine was substituted for each of 4 of the basic residues and 1 hydrophobic residue from the LPS-binding region of BPI and 4 hydrophobic residues from the beta-turn-inducing regions flanking the LPS-binding region on the carboxy-terminal side. Intriguingly, these last 4 substitutions resulted in peptides that exhibited increased bactericidal activity compared to betapep25. CONCLUSIONS: These results demonstrate the importance of both cationic and hydrophobic amino acid residues to bactericidal and endotoxin-neutralizing activities. These perturbations of biological activity should be considered in the design of synthetic peptide endotoxin antagonists.     

Laparotomy potentiates cytokine release and impairs pulmonary function after hemorrhage and resuscitation in mice

BACKGROUND: The two-hit theory has emerged as a mechanism to explain the development of organ failure after traumatic injury. We evaluated the effects of exploratory laparotomy (EL) as a second hit on mice after hemorrhage and resuscitation (H/R). Our hypothesis was that mice exposed to prior H/R would demonstrate more evidence of acute lung injury (ALI), as well as an augmented cytokine response, than mice exposed to H/R or EL alone. METHODS: Three groups of mice were examined. Mice undergoing H/R alone were labeled as the H/R group. Mice undergoing sham H/R (cannulation but no hemorrhage), followed 5 days later by EL, were labeled as the EL group; and mice undergoing H/R, followed 5 days later by an EL, were labeled as the H/R + EL, or two-hit, group. Respiratory function was determined by using whole-body plethysmography and lung gas diffusion. Serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were assayed at 1 and 4 hours after the injury stimuli. RESULTS: Evaluation of the change in pulmonary function after 24 hours demonstrated that EL alone induces a significant decrease in pulmonary function, whereas two-hit mice did not exhibit a potentiated response. Alveolar function was significantly degraded in the EL group compared with all other groups (p < 0.0001). TNF-alpha did not change after any injury at any time. However, evaluation of IL-6 levels demonstrated a substantial increase after H/R, EL, and H/R + EL compared with baseline and at 1 hour. Comparison of the three groups at 4 hours did not demonstrate any differences in serum concentrations of IL-6. Histologic evaluation lungs demonstrated that the most severe lung injury was seen in the EL mice. CONCLUSION: It would appear that serum TNF-alpha has little impact on the pathogenesis of ALI after EL, whereas serum IL-6 may be more important. Exploratory laparotomy resulted in a significant change in pulmonary function. Contrary to our initial hypothesis, two-hit mice did not demonstrate more evidence of ALI and, in fact, demonstrated less lung injury than EL mice.     

大鼠脑出血后血肿周围蛋白酶激活受体-1的表达及其机制

目的探讨脑出血后血肿周围蛋白酶激活受体-1(PAR-1)表达的情况及其与凝血酶的关系。方法胶原酶Ⅶ型、凝血酶(TM)、水蛭素分别脑内立体定向注射制作动物模型,应用RT- PCR技术检测PAR-1 mRNA表达,应用免疫组织化学技术检测PAR-1蛋白表达。结果脑出血后6 h PAR-1mRNA及蛋白表达分别为0．802±0．143和19．65±1．12,与对照组比较明显增加(P< 0．05),24 h分别为1．825±0．214和33．34±1．09(P<0．01),持续至72 h(P<0．01),然后逐渐减少,96 h时已恢复正常。凝血酶脑内注射后PAR-1 mRNA及蛋白表达的动态变化趋势与脑出血组相似,凝血酶组PAR-1 mRNA及蛋白表达在各个时间点与脑出血组比较差异均无统计学意义(P>0．05)。水蛭素组PAR-1 mRNA及蛋白表达与对照组比较差异无统计学意义(P>0．05)。结论脑出血后血肿周围PAR-1 mRNA和蛋白表达明显增加,可能是凝血酶直接作用的结果。PAR-1表达上调可能参与了脑出血后凝血酶神经毒性损伤过程。     

血管紧张素Ⅱ对大鼠肺微血管内皮细胞APJ mRNA和Apelin mRNA表达的影响

目的 探讨血管紧张素Ⅱ(AngⅡ)对肺微血管内皮细胞APJ mRNA和Apelin mRNA表达的影响.方法 培养并鉴定24 h新生SD大鼠肺微血管内皮细胞,取2～4代培养细胞,细胞密度5×105个/ml,培养至80%融合后,进行药物干预.第一部分实验分为5组(n=4):加入AngⅡ至终浓度分别为10-9 mol/L(Ⅱ组)、10-1 mol/L(Ⅲ组)、10-7mol/L(Ⅳ组)、10-6 mol/L(Ⅴ组),Ⅰ组不加入AngⅡ,24 h后采用RT-PCR技术测定Apelin mRNA和APJ mRNA的表达水平.第二部分实验分为6组(n=4):加入AngⅡ至终浓度为10-7mol/L,分别在孵育即刻(Ⅰ组)、1 h(Ⅱ组)、6 h(Ⅲ组)、12 h(Ⅳ组)、24 h(Ⅴ组)、48 h(Ⅵ组)时,采用RT-PCR技术测定Apelin mRNA和APJ mRNA的表达水平.结果 第一部分实验结果:与Ⅰ组比较,Ⅱ组Apelin mRNA表达上调,APJ mRNA表达下调,Ⅲ组～V组Apelin mRNA和APJ mRNA表达下调(P<0.05或0.01),与Ⅱ组比较,Ⅲ组～Ⅴ组Apelin mRNA和APJ mRNA表达下调,呈浓度依赖性(P<0.01).第二部分实验结果:Apelin mRNA在10-7mol/L AngⅡ孵育6 h内表达上调,孵育1 h时达高峰(P<0.05或0.01),孵育6 h后Apelin mRNA表达下调,且呈时间依赖性(P<0.01);而APJ mRNA在孵育12 h后呈时间依赖性持续下调(P<0.01).结论 AngⅡ呈浓度和时间依赖性地下调Apelin mRNA和APJ mRNA的表达,可能是其参与肺微血管内皮细胞损伤的发生机制.