全氟化碳吸入对脂多糖诱导大鼠急性肺损伤的保护作用及机制研究
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摘要
背景与目的:
急性肺损伤(ALI)是由严重感染、创伤、休克等因素引起的以炎症反应和肺泡毛细血管膜损伤为主要病理改变,以进行性呼吸困难和顽固性低氧血症为特征的危重症,病情凶险,死亡率高。内毒素是ALI的首要致病因素,肺微血管内皮细胞(PMVEC)及其表达的细胞间黏附分子-1(ICAM-1)在内毒素性ALI病理过程中发挥着关键作用。目前临床治疗尚未取得明显进展,呼吸支持仍是主要治疗手段之一。采用全氟化碳(PFC)液体通气,尤其是雾化吸入治疗ALI较常规机械通气(CMV)有明显优势。然而,其机制尚未完全阐明,特别是PFC的抗炎效应仍有争议,对PMVEC的保护作用及机制有待进一步探讨。为此,本课题拟从器官水平观察PFC雾化吸入对内毒素性ALI大鼠肺微血管通透性的影响,从细胞水平检测PFC作用下脂多糖(LPS)诱导大鼠PMVEC表达ICAM-1的变化,从分子水平探讨PFC对LPS激活大鼠肺微血管内皮细胞Toll样受体4(TLR4)信号转导通路的干预作用,旨在证实PFC的生物学抗炎效应,阐明PFC保护PMVEC的机理,从抗炎角度揭示PFC雾化吸入治疗内毒素性ALI的机制,为临床应用PFC雾化吸入治疗内毒素性ALI提供更坚实的理论基础和实验依据。
方法:
1.采用颈静脉注射LPS的方法复制大鼠内毒素性ALI模型,按随机原则将64只SD大鼠分为正常对照组(N)、脂多糖致伤组(LPS)、常规机械通气治疗组(CMV)和全氟化碳雾化吸入治疗组(PFC)。LPS组大鼠经颈静脉注射6mg/kg LPS致肺损伤;两治疗组大鼠在造模成功后给予常规机械通气,其中PFC组联合应用PFC 10ml/kg/h雾化吸入。通气2h并续观6h后,采集各组大鼠标本,测定动脉血氧分压(PaO2)和肺组织湿干重比(W/D),采用伊文思蓝示踪法检测肺微血管通透性、邻连茴香胺法测定肺组织MPO活性、RT-PCR法和Western Blot法分别检测肺组织ICAM-1 mRNA和蛋白表达水平,并进行肺组织病理形态学观察。
2.采用外周肺组织贴块法培养大鼠PMVEC,抽取组织块进行切片观察,以大鼠肺动脉平滑肌细胞和人脐静脉内皮细胞为对照,对培养细胞进行CD34、植物凝集素BSI、Ⅷ因子相关抗原免疫细胞化学染色鉴定,通过光镜和透射电镜观察细胞形态和超微结构。
3.将对数生长期的大鼠PMVEC接种于Transwell小室透明聚乙烯膜上,在内外室中加入培养液继续培养,融合成致密单层后,随机分为空白对照组(C)、PFC干预组(F)、LPS刺激组(L)、LPS刺激+PFC干预组(LF)。以处理0h为空白对照组,F组在Transwell外室中加入PFC进行干预,L组在Transwell内室中给予100ng/ml LPS刺激细胞,LF组在Transwell内室和外室中分别施加100ng/ml LPS和PFC进行共孵育。各处理组细胞孵育3、8、24h后,通过光镜和MTT法分别观测F组细胞形态和活力,采用RT-PCR法和流式细胞术(FCM)分别检测各组细胞ICAM-1 mRNA和蛋白表达水平。
4.将大鼠PMVEC培养于Transwell小室,融合成致密单层后,按上述方法分组和处理细胞,24h后采用FCM检测C组、F组细胞TLR4蛋白表达水平及FITC-LPS结合量,应用Western Blot和EMSA法分别测定C组、L组、LF组细胞TRAF6、磷酸化p38MAPK蛋白表达量和NF-κB活化水平。
结果:
1.与正常对照组比较,LPS致伤组大鼠的PaO2下降、W/D比值增大、肺微血管通透性增强、肺组织MPO活性增高,肺组织ICAM-1 mRNA和蛋白表达水平上调,肺组织病理形态学积分增加(P﹤0.01);CMV治疗组大鼠的上述指标与LPS致伤组比较无显著性差异(P﹥0.05);PFC雾化吸入治疗组则较LPS致伤组和CMV治疗组均有显著改善(P﹤0.01-0.05)。
2.组织块法培养大鼠PMVEC的综合鉴定结果显示:组织块源于外周肺组织,CD34免疫细胞化学染色阳性,植物凝集素BSI结合试验阳性,而Ⅷ因子相关抗原染色阴性,透射电镜未见Weibel-Palade小体。
3.正常大鼠PMVEC经PFC干预3、8、24h后,细胞形态、细胞活力、ICAM-1 mRNA和蛋白表达水平与C组比较无显著性差异(P﹥0.05);L组给予LPS刺激3、8、24h后,各时相点细胞的ICAM-1 mRNA和蛋白表达量均较C组显著增高(P﹤0.01-0.05);LF组细胞经LPS刺激和PFC干预后,与L组同时相点比较,3h组ICAM-1 mRNA和蛋白表达水平无明显差异(P﹥0.05),但8h组和24h组均有显著降低(P﹤0.01-0.05)。
4.流式细胞术检测结果发现,F组细胞TLR4蛋白表达水平及FITC-LPS结合量与C组比较无显著性差异(P﹥0.05);Western Blot和EMSA测定结果显示,L组细胞TRAF6的蛋白表达量、p38MAPK的磷酸化水平及NF-κB的结合活性均较C组显著增高(P﹤0.01-0.05);与L组比较,LF组细胞TRAF6蛋白表达量无显著性差异(P﹥0.05),但p38MAPK的磷酸化水平和NF-κB的结合活性均有显著降低(P﹤0.05)。
结论:
1.全氟化碳雾化吸入对LPS诱导的大鼠ALI具有保护作用,可明显减轻肺部炎症反应、改善肺微血管通透性,下调肺组织ICAM-1表达、抑制PMN肺内聚集是其保护机制之一。
2.外周肺组织贴块法是一种简便、经济、有效的大鼠PMVEC体外培养方法,Ⅷ因子相关抗原和Weibel-Palade小体并非鉴定大鼠PMVEC的理想指标,联合应用外周肺组织切片、CD34和植物凝集素BSI三指标为组织块法培养的大鼠PMVEC提供了一个简单易行、合理、可靠的综合鉴定方案。
3.全氟化碳不影响正常大鼠PMVEC的细胞形态和细胞活力,可通过生物学抗炎效应下调LPS诱导的肺微血管内皮细胞ICAM-1表达,进而保护PMVEC。
4.全氟化碳作用于LPS刺激的大鼠PMVEC后,既未影响细胞外LPS与TLR4的亲和,也不阻断TRAF6上游的TLR4细胞内信号转导通路,而是通过抑制TRAF6下游NF-κB和p38MAPK的活化而干预LPS诱导PMVEC表达ICAM-1。
Background and Objective
Acute lung injury (ALI) is a critical clinical syndrome with progressive dyspnea, refractory hypoxemia and high mortality, which pathological features mainly include inflammatory reaction and alveolar-capillary membrane injury result from severe infection, trauma, shock, etc.Endotoxima is a key etiological factor for ALI. Pulmonary microvascular endothelial cells (PMVEC) and overexpression of intercellular adhesion molecular-1 (ICAM-1) play key roles in the pathogenesis of lipopolysaccharide (LPS)-induced ALI. So far, the therapeutic methods for ALI have not been improved. Respiratory support is still one of major methods for treating ALI. The studies have shown that liquid ventilation, especially aerosol therapy with perfluorocarbon (PFC) has more advantages than conventional mechanical ventilation (CMV) in the treatment of ALI. However, its mechanism has not been completely elucidated. Especially the anti-inflammatory effect of PFC is still controversial and the protective effects of PFC on PMVEC remain unclear. Therefor, in this study we will observe the effects of aerosolized perfluorocarbon on pulmonary microvascular permeability in the a model of LPS-induced ALI in vivo, evaluate the impact of PFC on LPS-induced ICAM-1 expression in cultured PMVECs of rat, and then investigate the influence of PFC on LPS-activated TLR4 signal transduction in vitro. The purposes of this study are to confirm whether there is anti-inflammatory effect of PFC for ALI, to elucidate the protective mechanism of PFC on PMVEC, and then reveal the anti-inflammatory mechanism of aerosolized PFC on LPS-induced ALI.
Methods
1. A rat model of ALI was established by jugular injection of LPS at a dose of 6mg/kg. Sixty-four SD rats were randomly divided into four goups: N, LPS, CMV and PFC. Group N remained untreated as normal control, group LPS was injured by LPS injection as ALI model. After establishment of lung injury by LPS, group CMV received the conventional mechanical ventilation, group PFC received both the conventional mechanical ventilation and inhalation of aerosolized PFC at a dose of 10ml/kg/h. After 2 hours treatment and another 6 hours observation, the artery blood and lung tissue were collected. The level of PaO2 in artery blood and the wet/dry ratio of lung tissue were measured. The pulmonary microvascular permeability and MPO activity of lung tissue were detected. The level of ICAM-1 mRNA and protein expression in lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot, respectively. The histological changes of lung tissue were observed under optical microscope.
2. Rat PMVECs were cultured by peripheral lung tissue-sticking method. Histological sections from peripheral lung tissue pieces used for cell culture were examined. Used rat pulmonary artery smooth muscle cells and human umbilical vein endothelial cells as control, CD34, lectin from Bandeiraea simplicifolia and factorⅧrelated antigen in the cultured cells were quantified by immunocytochemical staining. In addition, the cell morphology and ultrastructure were observed with inverted optical microscope and transmission electron microscope respectively.
3. Rat PMVECs cultured by peripheral lung tissue-sticking method were seeded in Transwell chambers until confluence, and divided randomly into 4 groups: group C remained untreated as blank control; group F was incubated with PFC; group L was incubated with 100ng/ml LPS; group LF was incubated with both 100ng/ml LPS and PFC. After treatment for 3, 8 and 24 hours, the cell morphology of group F was observed with optical microscope. Then the cells of each group were harvested, respectively. The cell activity in group F was determined by MTT assay. The mRNA and protein expression levels of ICAM-1 in each group were detected by RT-PCR and flow cytometry (FCM), respectively.
4. Rat PMVECs were cultured, grouped and treated as mentioned above. Twenty-four hours later, the level of TLR4 protein expression and its affinity with FITC-LPS in group F and goup C were detected by FCM. The level of TRAF6 protein expression and the activities of p38MAPK and NF-κB were determined by Western Blot and EMSA.
Results
1. Compared with group N, PaO2 decreased and W/D ratio, pulmonary microvascular permeability, MPO activity, ICAM-1 mRNA and protein expression, pathomorphological score of lung tissue in group LPS increased significantly (P<0.01).There was no significant difference between group CMV and group LPS (P>0.05). However, compared with group CMV and group LPS, the above indexes in group PFC improved significantly (P<0.01-0.05).
2. Histological sections showed that tissue pieces were scissored from periphery lung lobes accurately. The cultured cells had features of binding lectin from Bandeiraea simplicifolia and positive immunocytochemical staining with CD34 antibody, but negative for factorⅧrelated antigen. Weibel-Palade bodies were not observed in the cultured cells.
3. The cell morphology and activity in normal rat PMVECs did not have significant change between before and after PFC treatment (P>0.05), despite at 3, 8 or 24 h. Both mRNA and protein expression levels of ICAM-1 had no significant difference between group F and group C (P>0.05). Group L had significantly higher the mRNA and protein expression levels of ICAM-1 in a time-dependent manner than group C (P<0.01-0.05). In comparison with group L, the mRNA and protein expression levels of ICAM-1 in group LF were not suppressed significantly by treatment at 3h-point (P>0.05), however, they markedly decreased at 8h-point and 24h-point (P<0.01-0.05).
4. The TLR4 protein concentration and FITC-LPS affinity on cell membrance had no significant difference between group N and group F (P>0.05). The levels of TRAF6 protein expression, the activity of p38MAPK and NF-κB in group L were significantly higher than those in group C (P<0.01-0.05). There was no significant defference in TRAF6 protein expression between group LF and group L. But compared with those in group L, the activity of p38MAPK and NF-κB in group LF decreased significantly (P<0.05).
Conclusion
1. Aerosol therapy with perfluorocarbon significantly protects lung from LPS-induced injury in rats by the suppression of intercellular adhesion molecular-1 expression and polymorphonuclear leukocyte accumulation in lung tissue and the subsequent attenuation of pulmonary microvascular permeability.
2. Peripheral lung tissue-sticking is a convenient, economical and efficacious method for rat PMVEC cultivation. FactorⅧrelated antigen and Weibel-Palade body are not ideal indexes for ascertaining rat PMVEC. The combination of peripheral lung tissue section, CD34 immunocytochemical staining and lectin from Bandeiraea simplicifolia binding assay is a simple and reasonable comprehensive method for identifying rat PMVECs.
3. The cell morphology and viability of normal rat PMVECs are not significantly impaired by PFC treatment Perfluorocarbon suppresses the mRNA and protein expression of ICAM-1 induced by LPS which suggests a potential mechanism that PFC protects PMVECs from LPS-induced injury via its biological anti-inflammatory effect.
4. The interaction between TLR4 and LPS on PMVECs surface membrane is not altered by PFC. The inhibition of PFC on TRAF6 and NF-κB activation seems to be the cause of attenuated ICAM-1 overexpression induced by LPS.
急性肺损伤(ALI)是由严重感染、创伤、休克等因素引起的以炎症反应和肺泡毛细血管膜损伤为主要病理改变,以进行性呼吸困难和顽固性低氧血症为特征的危重症,病情凶险,死亡率高。内毒素是ALI的首要致病因素,肺微血管内皮细胞(PMVEC)及其表达的细胞间黏附分子-1(ICAM-1)在内毒素性ALI病理过程中发挥着关键作用。目前临床治疗尚未取得明显进展,呼吸支持仍是主要治疗手段之一。采用全氟化碳(PFC)液体通气,尤其是雾化吸入治疗ALI较常规机械通气(CMV)有明显优势。然而,其机制尚未完全阐明,特别是PFC的抗炎效应仍有争议,对PMVEC的保护作用及机制有待进一步探讨。为此,本课题拟从器官水平观察PFC雾化吸入对内毒素性ALI大鼠肺微血管通透性的影响,从细胞水平检测PFC作用下脂多糖(LPS)诱导大鼠PMVEC表达ICAM-1的变化,从分子水平探讨PFC对LPS激活大鼠肺微血管内皮细胞Toll样受体4(TLR4)信号转导通路的干预作用,旨在证实PFC的生物学抗炎效应,阐明PFC保护PMVEC的机理,从抗炎角度揭示PFC雾化吸入治疗内毒素性ALI的机制,为临床应用PFC雾化吸入治疗内毒素性ALI提供更坚实的理论基础和实验依据。
方法:
1.采用颈静脉注射LPS的方法复制大鼠内毒素性ALI模型,按随机原则将64只SD大鼠分为正常对照组(N)、脂多糖致伤组(LPS)、常规机械通气治疗组(CMV)和全氟化碳雾化吸入治疗组(PFC)。LPS组大鼠经颈静脉注射6mg/kg LPS致肺损伤;两治疗组大鼠在造模成功后给予常规机械通气,其中PFC组联合应用PFC 10ml/kg/h雾化吸入。通气2h并续观6h后,采集各组大鼠标本,测定动脉血氧分压(PaO2)和肺组织湿干重比(W/D),采用伊文思蓝示踪法检测肺微血管通透性、邻连茴香胺法测定肺组织MPO活性、RT-PCR法和Western Blot法分别检测肺组织ICAM-1 mRNA和蛋白表达水平,并进行肺组织病理形态学观察。
2.采用外周肺组织贴块法培养大鼠PMVEC,抽取组织块进行切片观察,以大鼠肺动脉平滑肌细胞和人脐静脉内皮细胞为对照,对培养细胞进行CD34、植物凝集素BSI、Ⅷ因子相关抗原免疫细胞化学染色鉴定,通过光镜和透射电镜观察细胞形态和超微结构。
3.将对数生长期的大鼠PMVEC接种于Transwell小室透明聚乙烯膜上,在内外室中加入培养液继续培养,融合成致密单层后,随机分为空白对照组(C)、PFC干预组(F)、LPS刺激组(L)、LPS刺激+PFC干预组(LF)。以处理0h为空白对照组,F组在Transwell外室中加入PFC进行干预,L组在Transwell内室中给予100ng/ml LPS刺激细胞,LF组在Transwell内室和外室中分别施加100ng/ml LPS和PFC进行共孵育。各处理组细胞孵育3、8、24h后,通过光镜和MTT法分别观测F组细胞形态和活力,采用RT-PCR法和流式细胞术(FCM)分别检测各组细胞ICAM-1 mRNA和蛋白表达水平。
4.将大鼠PMVEC培养于Transwell小室,融合成致密单层后,按上述方法分组和处理细胞,24h后采用FCM检测C组、F组细胞TLR4蛋白表达水平及FITC-LPS结合量,应用Western Blot和EMSA法分别测定C组、L组、LF组细胞TRAF6、磷酸化p38MAPK蛋白表达量和NF-κB活化水平。
结果:
1.与正常对照组比较,LPS致伤组大鼠的PaO2下降、W/D比值增大、肺微血管通透性增强、肺组织MPO活性增高,肺组织ICAM-1 mRNA和蛋白表达水平上调,肺组织病理形态学积分增加(P﹤0.01);CMV治疗组大鼠的上述指标与LPS致伤组比较无显著性差异(P﹥0.05);PFC雾化吸入治疗组则较LPS致伤组和CMV治疗组均有显著改善(P﹤0.01-0.05)。
2.组织块法培养大鼠PMVEC的综合鉴定结果显示:组织块源于外周肺组织,CD34免疫细胞化学染色阳性,植物凝集素BSI结合试验阳性,而Ⅷ因子相关抗原染色阴性,透射电镜未见Weibel-Palade小体。
3.正常大鼠PMVEC经PFC干预3、8、24h后,细胞形态、细胞活力、ICAM-1 mRNA和蛋白表达水平与C组比较无显著性差异(P﹥0.05);L组给予LPS刺激3、8、24h后,各时相点细胞的ICAM-1 mRNA和蛋白表达量均较C组显著增高(P﹤0.01-0.05);LF组细胞经LPS刺激和PFC干预后,与L组同时相点比较,3h组ICAM-1 mRNA和蛋白表达水平无明显差异(P﹥0.05),但8h组和24h组均有显著降低(P﹤0.01-0.05)。
4.流式细胞术检测结果发现,F组细胞TLR4蛋白表达水平及FITC-LPS结合量与C组比较无显著性差异(P﹥0.05);Western Blot和EMSA测定结果显示,L组细胞TRAF6的蛋白表达量、p38MAPK的磷酸化水平及NF-κB的结合活性均较C组显著增高(P﹤0.01-0.05);与L组比较,LF组细胞TRAF6蛋白表达量无显著性差异(P﹥0.05),但p38MAPK的磷酸化水平和NF-κB的结合活性均有显著降低(P﹤0.05)。
结论:
1.全氟化碳雾化吸入对LPS诱导的大鼠ALI具有保护作用,可明显减轻肺部炎症反应、改善肺微血管通透性,下调肺组织ICAM-1表达、抑制PMN肺内聚集是其保护机制之一。
2.外周肺组织贴块法是一种简便、经济、有效的大鼠PMVEC体外培养方法,Ⅷ因子相关抗原和Weibel-Palade小体并非鉴定大鼠PMVEC的理想指标,联合应用外周肺组织切片、CD34和植物凝集素BSI三指标为组织块法培养的大鼠PMVEC提供了一个简单易行、合理、可靠的综合鉴定方案。
3.全氟化碳不影响正常大鼠PMVEC的细胞形态和细胞活力,可通过生物学抗炎效应下调LPS诱导的肺微血管内皮细胞ICAM-1表达,进而保护PMVEC。
4.全氟化碳作用于LPS刺激的大鼠PMVEC后,既未影响细胞外LPS与TLR4的亲和,也不阻断TRAF6上游的TLR4细胞内信号转导通路,而是通过抑制TRAF6下游NF-κB和p38MAPK的活化而干预LPS诱导PMVEC表达ICAM-1。
Background and Objective
Acute lung injury (ALI) is a critical clinical syndrome with progressive dyspnea, refractory hypoxemia and high mortality, which pathological features mainly include inflammatory reaction and alveolar-capillary membrane injury result from severe infection, trauma, shock, etc.Endotoxima is a key etiological factor for ALI. Pulmonary microvascular endothelial cells (PMVEC) and overexpression of intercellular adhesion molecular-1 (ICAM-1) play key roles in the pathogenesis of lipopolysaccharide (LPS)-induced ALI. So far, the therapeutic methods for ALI have not been improved. Respiratory support is still one of major methods for treating ALI. The studies have shown that liquid ventilation, especially aerosol therapy with perfluorocarbon (PFC) has more advantages than conventional mechanical ventilation (CMV) in the treatment of ALI. However, its mechanism has not been completely elucidated. Especially the anti-inflammatory effect of PFC is still controversial and the protective effects of PFC on PMVEC remain unclear. Therefor, in this study we will observe the effects of aerosolized perfluorocarbon on pulmonary microvascular permeability in the a model of LPS-induced ALI in vivo, evaluate the impact of PFC on LPS-induced ICAM-1 expression in cultured PMVECs of rat, and then investigate the influence of PFC on LPS-activated TLR4 signal transduction in vitro. The purposes of this study are to confirm whether there is anti-inflammatory effect of PFC for ALI, to elucidate the protective mechanism of PFC on PMVEC, and then reveal the anti-inflammatory mechanism of aerosolized PFC on LPS-induced ALI.
Methods
1. A rat model of ALI was established by jugular injection of LPS at a dose of 6mg/kg. Sixty-four SD rats were randomly divided into four goups: N, LPS, CMV and PFC. Group N remained untreated as normal control, group LPS was injured by LPS injection as ALI model. After establishment of lung injury by LPS, group CMV received the conventional mechanical ventilation, group PFC received both the conventional mechanical ventilation and inhalation of aerosolized PFC at a dose of 10ml/kg/h. After 2 hours treatment and another 6 hours observation, the artery blood and lung tissue were collected. The level of PaO2 in artery blood and the wet/dry ratio of lung tissue were measured. The pulmonary microvascular permeability and MPO activity of lung tissue were detected. The level of ICAM-1 mRNA and protein expression in lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot, respectively. The histological changes of lung tissue were observed under optical microscope.
2. Rat PMVECs were cultured by peripheral lung tissue-sticking method. Histological sections from peripheral lung tissue pieces used for cell culture were examined. Used rat pulmonary artery smooth muscle cells and human umbilical vein endothelial cells as control, CD34, lectin from Bandeiraea simplicifolia and factorⅧrelated antigen in the cultured cells were quantified by immunocytochemical staining. In addition, the cell morphology and ultrastructure were observed with inverted optical microscope and transmission electron microscope respectively.
3. Rat PMVECs cultured by peripheral lung tissue-sticking method were seeded in Transwell chambers until confluence, and divided randomly into 4 groups: group C remained untreated as blank control; group F was incubated with PFC; group L was incubated with 100ng/ml LPS; group LF was incubated with both 100ng/ml LPS and PFC. After treatment for 3, 8 and 24 hours, the cell morphology of group F was observed with optical microscope. Then the cells of each group were harvested, respectively. The cell activity in group F was determined by MTT assay. The mRNA and protein expression levels of ICAM-1 in each group were detected by RT-PCR and flow cytometry (FCM), respectively.
4. Rat PMVECs were cultured, grouped and treated as mentioned above. Twenty-four hours later, the level of TLR4 protein expression and its affinity with FITC-LPS in group F and goup C were detected by FCM. The level of TRAF6 protein expression and the activities of p38MAPK and NF-κB were determined by Western Blot and EMSA.
Results
1. Compared with group N, PaO2 decreased and W/D ratio, pulmonary microvascular permeability, MPO activity, ICAM-1 mRNA and protein expression, pathomorphological score of lung tissue in group LPS increased significantly (P<0.01).There was no significant difference between group CMV and group LPS (P>0.05). However, compared with group CMV and group LPS, the above indexes in group PFC improved significantly (P<0.01-0.05).
2. Histological sections showed that tissue pieces were scissored from periphery lung lobes accurately. The cultured cells had features of binding lectin from Bandeiraea simplicifolia and positive immunocytochemical staining with CD34 antibody, but negative for factorⅧrelated antigen. Weibel-Palade bodies were not observed in the cultured cells.
3. The cell morphology and activity in normal rat PMVECs did not have significant change between before and after PFC treatment (P>0.05), despite at 3, 8 or 24 h. Both mRNA and protein expression levels of ICAM-1 had no significant difference between group F and group C (P>0.05). Group L had significantly higher the mRNA and protein expression levels of ICAM-1 in a time-dependent manner than group C (P<0.01-0.05). In comparison with group L, the mRNA and protein expression levels of ICAM-1 in group LF were not suppressed significantly by treatment at 3h-point (P>0.05), however, they markedly decreased at 8h-point and 24h-point (P<0.01-0.05).
4. The TLR4 protein concentration and FITC-LPS affinity on cell membrance had no significant difference between group N and group F (P>0.05). The levels of TRAF6 protein expression, the activity of p38MAPK and NF-κB in group L were significantly higher than those in group C (P<0.01-0.05). There was no significant defference in TRAF6 protein expression between group LF and group L. But compared with those in group L, the activity of p38MAPK and NF-κB in group LF decreased significantly (P<0.05).
Conclusion
1. Aerosol therapy with perfluorocarbon significantly protects lung from LPS-induced injury in rats by the suppression of intercellular adhesion molecular-1 expression and polymorphonuclear leukocyte accumulation in lung tissue and the subsequent attenuation of pulmonary microvascular permeability.
2. Peripheral lung tissue-sticking is a convenient, economical and efficacious method for rat PMVEC cultivation. FactorⅧrelated antigen and Weibel-Palade body are not ideal indexes for ascertaining rat PMVEC. The combination of peripheral lung tissue section, CD34 immunocytochemical staining and lectin from Bandeiraea simplicifolia binding assay is a simple and reasonable comprehensive method for identifying rat PMVECs.
3. The cell morphology and viability of normal rat PMVECs are not significantly impaired by PFC treatment Perfluorocarbon suppresses the mRNA and protein expression of ICAM-1 induced by LPS which suggests a potential mechanism that PFC protects PMVECs from LPS-induced injury via its biological anti-inflammatory effect.
4. The interaction between TLR4 and LPS on PMVECs surface membrane is not altered by PFC. The inhibition of PFC on TRAF6 and NF-κB activation seems to be the cause of attenuated ICAM-1 overexpression induced by LPS.
引文
1. Tamura DY, Moore EE, Johnson JL, et al. p38 mitogen-activated protein kinase inhibition attenuates intercellular adhesion molecule-1 up-regulation on human pulmonary microvascular endothelial cells[J]. Surgery, 1998, 124(2): 403-407.
2. Christman JW, Sadikot RT, Blackwell TS. The role of nuclear factor-κB in pulmonary diseases[J]. Chest, 2000, 117(5): 1482-1487.
3. Carden D, Xiao F, Moak C, et al. Neutrophil elastase promotes lung microvascular injury and proteolysis of endothelial cadherins[J]. Am J Physiol, 1998, 275(2 Pt 2): H385-392.
4. Wang Q, Yerukhimovich M, Gaarde WA, et al. MKK3 and -6-dependent activation of p38alpha MAP kinase is required for cytoskeletal changes in pulmonary microvascular endothelial cells induced by ICAM-1 ligation[J]. Am J Physiol Lung Cell Mol Physiol, 2005, 288(2): L359-369.
5. Hübler M, Souders JE, Shade ED, et al. Effects of vaporized perfluorocarbon on pulmonary blood flow and ventilation perfusion distribution in a model of acute respiratory distress syndrome[J]. Anesthesiology, 2001, 95(6): 1414-1421.
6. Bleyl JU, Ragaller M, Tsch? U, et al. Changes in pulmonary function and oxygenation during application of perfluorocarbon vapor in healthy and oleic acid-injured animals[J]. Crit Care Med, 2002, 30(6): 1340-1347.
7. van Eeden SF, Klut ME, Leal MA, et al. Partial liquid ventilation with perfluorocarbon in acute lung injury light and transmission electron microscopy studies[J]. Am J Respir Cell Mol Biol, 2000, 22(4): 441-450.
8. Colton DM, Till GO, Johnson KJ, et al. Partial liquid ventilation decreases albumin leak in the setting of acute lung injury[J]. J Crit Care, 1998, 13(3): 136-139.
9. Kawamae K, Pristine G, Chiumello D, et al. Partial liquid ventilation decreases serum tumor necrosis factor-alpha concentrations in a rat acid aspiration lung injury model[J]. Crit Care Med, 2000, 28(2): 479-483.
10.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary iInflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
11.Heard SO, Puyana JC. The anti-inflammatory effects of perfluorocarbons: let's get physical[J]. Crit Care Med, 2000, 28(4): 1241-1242.
12.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha- induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
13.Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
14.Cox C, Stavis RL, Wolfson MR, et al. Long-term tidal liquid ventilation in premature lambs: physiologic, biochemical and histological correlates[J]. Biol Neonate, 2003, 84(3): 232-242.
15.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
16.Rotta AT, Steinhorn DM. Partial liquid ventilation reduces pulmonary neutrophil accumulation in an experimental model of systemic endotoxemia and acute lung injury[J]. Crit Care Med, 1998, 26(10): 1707-1715.
17.Ellena JF, Obraztsov VV, Cumbea VL, et al. Perfluorooctyl bromide has limited membrane solubility and is located at the bilayer center. Locating small molecules in lipid bilayers through paramagnetic enhancements of NMR relaxation[J]. J Med Chem, 2002, 45(25): 5534-5542.
18.Nakstad B, Wolfson MR, Shaffer TH, et al. Perfluorochemical liquids modulate cell-mediated inflammatory responses[J]. Crit Care Med, 2001, 29(9): 1731-1737.
19.Fernandez R, Sarma V, Younkin E, et al. Exposure to perflubron is associated with decreased Syk phosphorylation in human neutrophils[J]. J Appl Physiol, 2001, 91(5): 1941-1947.
20.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
21.Colton DM, Till GO, Johnson KJ, et al. Neutrophil accumulation is reduced during partial liquid ventilation[J]. Crit Care Med, 1998, 26(10): 1716-1724.
22.Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
23.Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
24.Jeng MJ, Kou YR, Sheu CC, et al. Effects of partial liquid ventilation with FC-77 on acute lung injury in newborn piglets[J]. Pediatr Pulmonol, 2002, 33(1): 12-21.
25.Suh GY, Chung MP, Park SJ, et al. Partial liquid ventilation with perfluorocarbon improves gas exchange and decreases inflammatory response in oleic acid-induced lung injury in beagles[J]. J Korean Med Sci, 1999, 14(6): 613-622.
26.Croce MA, Fabian TC, Patton JH Jr, et al. Partial liquid ventilation decreases the inflammatory response in the alveolar environment of trauma patients[J]. J Trauma, 1998, 45(2): 273-280.
27.Rotta AT, Gunnarsson B, Hernan LJ, et al. Partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids[J]. Crit Care Med, 2000, 28(1): 202-208.
28.Dani C, Costantino ML, Martelli E, et al. Perfluorocarbons attenuate oxidative lung damage[J]. Pediatr Pulmonol, 2003, 36(4): 322-329.
29.Hirayama Y, Hirasawa H, Oda S, et al. Partial liquid ventilation with FC-77 suppresses the release of lipid mediators in rat acute lung injury model[J]. Crit Care Med, 2004, 32(10): 2085-2089.
30.Hirschl RB, Conrad S, Kaiser R, et al. Partial liquid ventilation in adult patients with ARDS: a multicenter phase I-II trial. Adult PLV Study Group[J]. Ann Surg, 1998, 228(5): 692-700.
31.Kandler MA, von der Hardt K, Schoof E, et al. Persistent improvement of gas exchange and lung mechanics by aerosolized perfluorocarbon[J]. Am J Respir Crit Care Med, 2001, 164(1): 31-35.
32.Ragaller M, Bleyl J, Tscho U, et al. Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury[J]. Intensive Care Med, 2001, 27(5): 889-897.
33.Thomas SR, Gradon L, Pratsinis SE, et al. Perfluorocarbon compound aerosols for delivery to the lung as potential 19F magnetic resonance reporters of regional pulmonary pO2[J]. Invest Radiol, 1997, 32(1): 29-38.
34.Hildebrand F, Pape HC, Harwood P, et al. Role of adhesion molecule ICAM in the pathogenesis of polymicrobial sepsis[J]. Exp Toxicol Pathol, 2005, 56(4-5): 281-290.
35.Reickert C, Pranikoff T, Overbeck M, et al. The pulmonary and systemic distribution and elimination of perflubron from adult patients treated with partial liquid ventilation[J]. Chest, 2001, 119(2): 515-522.
36.Varani J, Hirschl RB, Dame M, et al. Perfluorocarbon protects lung epithelial cells from neutrophil-mediated injury in an in vitro model of liquid ventilation therapy[J]. Shock, 1996, 6(5): 339-344.
37.Chi JT, Chang HY, Haraldsen G, et al. Endothelial cell diversity revealed by global expression profiling[J]. Proc Natl Acad Sci U S A, 2003, 100(19): 10623-10628.
38.King J, Hamil T, Creighton G, et al. Structural and functional characteristics of lung macro- and microvascular endothelial cell phenotypes[J]. Microvasc Res, 2004, 67(2): 139-151.
39.Medhora M, Daniels J, Mundey K, et al. Epoxygenase-driven angiogenesis in human lung microvascular endothelial cells[J]. Am J Physiol Heart Circ Physiol, 2003, 284(1): H215-H224.
40.Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J].Crit Care Med, 1997, 25(12): 2045-2047.
41.Koch T, Ragaller M, Haufe D, et al. Perfluorohexane attenuates proinflammatory and procoagulatory response of activated monocytes and alveolar macrophages[J]. Anesthesiology, 94(1): 101-109.
42.Smith TM, Steinhorn DM, Thusu K, et al. A liquid perfluorochemical decreases the in vitro production of reactive oxygen species by alveolar macrophages[J]. Crit Care Med, 1995, 23(9): 1533-1539.
43.Palsson-McDermott EM, O'Neill LA. Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4[J]. Immunology, 2004, 113(2): 153-162.
44.Poltorak A, He X, Smirnova I, et al. Defective LPS signaling in C3H/HeJ andC57BL/10ScCr mice: mutations in Tlr4 gene[J]. Science, 1998, 282(5396): 2085-2088.
45.Underhill DM, Ozinsky A. Toll-like receptors: key mediators of microbe detection[J]. Curr Opin Immunol, 2002, 14(1): 103-110.
46.Medzhitov R, Preston-Hurlburt P, Janeway CA Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J]. Nature, 1997, 388(6640): 394-397.
47.Nagai Y, Akashi S, Nagafuku M, et al. Essential role of MD-2 in LPS responsiveness and TLR4 distribution[J]. Nat Immunol, 2002, 3(7): 667-672.
48.Haufe D, Luther T, Kotzsch M, et al. Perfluorocarbon attenuates response of concanavalin A-stimulated mononuclear blood cells without altering ligand-receptor interaction[J]. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1): L210-L216.
49.Kobayashi T. Walsh MC. Choi Y. The role of TRAF6 in signal transduction and the immune response[J]. Microbes Infect, 2004, 6(14): 1333-1338.
50.Lomaga MA, Yeh WC, Sarosi I, et al. TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling[J]. Genes Dev, 1999, 13(8): 1015-1024.
1. Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
2. Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
3. Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
4. Clark LC, Gollan F. Survival of mammals breathing organic liquid sequilibrated with oxygenat at mospheric pressure[J].Science, 1966, 152(730): 1755-1756.
5. Fuhrman BP, Paczan PR, DeFrancisis M. Perfluorocarbon-associated gas exchange[J]. Crit Care Med, 1991, 19(5): 712-722.
6. Hirschl RB, Pranikoff T, Wise C, et al. Initial experience with partial liquid ventilation in adult patients with the acute respiratory distress syndrome[J]. JAMA, 1996, 275(5): 383-389.
7. Bleyl JU, Ragaller M, Tscho U, et al. Vaporized perfluorocarbon improves oxygenation and pulmonary function in an ovine model of acute respiratory distress syndrome[J]. Anesthesiology, 1999, 91(2): 461-469.
8. Thomas SR, Gradon L, Pratsinis SE, et al. Perfluorocarbon compound aerosols for delivery to the lung as potential 19F magnetic resonance reporters of regional pulmonary pO2[J]. Invest Radiol, 1997, 32(1): 29-38.
9. Thomas SH, O'Doherty MJ, Page CJ, et al. Delivery of ultrasonic nebulized aerosols to a lung model during mechanical ventilation. Am Rev Respir Dis, 1993, 148(4 Pt 1): 872-877.
10.Rudiger M, Gregor T, Burkhardt W, et al. Perfluorocarbon species and nebulizer type influence aerosolization rate and particle size of perfluorocarbon aerosol[J]. J Crit Care, 2004, 19(1): 42-47.
11.Gregor T, Schmalisch G, Burkhardt W, et al. Aerosolization of perfluorocarbons during mechanical ventilation: an in vitro study[J]. Intensive Care Med, 2003, 29(8): 1354-1360.
12.Ragaller M, Bleyl J, Tscho U, et al. Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury[J]. Intensive Care Med, 2001, 27(5): 889-897.
13.Kandler MA, von der Hardt K, Schoof E, et al. Persistent improvement of gas exchange and lung mechanics by aerosolized perfluorocarbon[J]. Am J Respir Crit Care Med, 2001, 164(1): 31-35.
14.Bleyl JU, Ragaller M, Tsch? U, et al. Changes in pulmonary function and oxygenation during application of perfluorocarbon vapor in healthy and oleic acid-injured animals[J]. Crit Care Med, 2002, 30(6): 1340-1347.
15.Hübler M, Souders JE, Shade ED, et al. Effects of vaporized perfluorocarbon on pulmonary blood flow and ventilation perfusion distribution in a model of acute respiratory distress syndrome[J]. Anesthesiology, 2001, 95(6): 1414-1421.
16.Hubler M, Heller AR, Bleyl JU, et al. Perfluorohexane vapor has only minor effects on spatial pulmonary blood flow distribution in isolated rabbit lungs[J]. Anesth Analg, 2005, 100(4): 1122-1128.
17.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
18.Schoof E, von der Hardt K, Kandler MA, et al. Aerosolized perfluorocarbon reduces adhesion molecule gene expression and neutrophil sequestration in acute respiratory distress[J]. Eur J Pharmacol, 2002, 457(2-3): 195-200.
19.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
20.Gama de Abreu M, Wilmink B, Hubler M, et al. Vaporized perfluorohexane attenuates ventilator-induced lung injury in isolated, perfused rabbit lungs[J]. Anesthesiology, 2005, 102(3): 597-605.
21.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha-induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
22.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
1. Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
2. Nakstad B, Wolfson MR, Shaffer TH, et al. Perfluorochemical liquids modulate cell-mediated inflammatory responses[J]. Crit Care Med, 2001, 29(9): 1731-1737.
3. Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
4. Cox C, Stavis RL, Wolfson MR, et al. Long-term tidal liquid ventilation in premature lambs: physiologic, biochemical and histological correlates[J]. Biol Neonate, 2003, 84(3): 232-242.
5. Reickert C, Pranikoff T, Overbeck M, et al. The pulmonary and systemic distribution and elimination of perflubron from adult patients treated with partial liquid ventilation[J]. Chest, 2001, 119(2): 515-522.
6. Ellena JF, Obraztsov VV, Cumbea VL, et al. Perfluorooctyl bromide has limited membrane solubility and is located at the bilayer center. Locating small molecules in lipid bilayers through paramagnetic enhancements of NMR relaxation[J]. J Med Chem, 2002, 45(25): 5534-5542.
7. Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
8. Koch T, Ragaller M, Haufe D, et al. Perfluorohexane attenuates proinflammatory and procoagulatory response of activated monocytes and alveolar macrophages[J]. Anesthesiology, 94(1): 101-109.
9. Quintel M, Heine M, Hirschl RB, et al. Effects of partial liquid ventilation on lung injury in a model of acute respiratory failure: a histologic and morphometric analysis[J]. Crit Care Med, 1998, 26(5): 833-843.
10.van Eeden SF, Klut ME, Leal MA, et al. Partial liquid ventilation with perfluorocarbonin acute lung injury light and transmission electron microscopy studies[J]. Am J Respir Cell Mol Biol, 2000, 22(4): 441-450.
11.Colton DM, Till GO, Johnson KJ, et al. Partial liquid ventilation decreases albumin leak in the setting of acute lung injury[J]. J Crit Care, 1998, 13(3): 136-139.
12.Lange NR, Kozlowski JK, Gust R, et al. Effect of partial liquid ventilation on pulmonary vascular permeability and edema after experimental acute lung injury[J]. Am J Respir Crit Care Med, 2000, 162(1): 271-277.
13.Rotta AT, Steinhorn DM. Partial liquid ventilation reduces pulmonary neutrophil accumulation in an experimental model of systemic endotoxemia and acute lung injury[J]. Crit Care Med, 1998, 26(10): 1707-1715.
14.Colton DM, Till GO, Johnson KJ, et al. Neutrophil accumulation is reduced during partial liquid ventilation[J]. Crit Care Med, 1998, 26(10): 1716-1724.
15.Nader ND, Knight PR, Davidson BA, et al. Systemic perfluorocarbons suppress the acute lung inflammation after gastric acid aspiration in rats[J]. Anesth Analg, 2000, 90(2): 356-361.
16.Suh GY, Chung MP, Park SJ, et al. Partial liquid ventilation with perfluorocarbon improves gas exchange and decreases inflammatory response in oleic acid-induced lung injury in beagles[J]. J Korean Med Sci, 1999, 14(6): 613-622.
17.Steinhorn DM, Papo MC, Rotta AT, et al. Liquid ventilation attenuates pulmonary oxidative damage[J]. J Crit Care, 1999, 14(1): 20-28.
18.Rotta AT, Gunnarsson B, Hernan LJ, et al. Partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids[J]. Crit Care Med, 2000, 28(1): 202-208.
19.Dani C, Costantino ML, Martelli E, et al. Perfluorocarbons attenuate oxidative lung damage[J]. Pediatr Pulmonol, 2003, 36(4): 322-329.
20.Hirayama Y, Hirasawa H, Oda S, et al. Partial liquid ventilation with FC-77 suppresses the release of lipid mediators in rat acute lung injury model[J]. Crit Care Med, 2004, 32(10): 2085-2089.
21.Schoof E, von der Hardt K, Kandler MA, et al. Aerosolized perfluorocarbon reduces adhesion molecule gene expression and neutrophil sequestration in acute respiratory distress[J]. Eur J Pharmacol, 2002, 457(2-3):195-200.
22.von der Hardt K, Kandler MA, Brenn G, et al. Comparison of aerosol therapy with different perfluorocarbons in surfactant-depleted animals[J]. Crit Care Med, 2004, 32(5): 1200-1206.
23.Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J].Crit Care Med, 1997, 25(12): 2045-2047.
24.Kawamae K, Pristine G, Chiumello D, et al. Partial liquid ventilation decreases serum tumor necrosis factor-alpha concentrations in a rat acid aspiration lung injury model[J]. Crit Care Med, 2000, 28(2): 479-483.
25.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
26.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
27.Croce MA, Fabian TC, Patton JH Jr, et al. Partial liquid ventilation decreases the inflammatory response in the alveolar environment of trauma patients[J]. J Trauma, 1998, 45(2): 273-280.
28.Haufe D, Luther T, Kotzsch M, et al. Perfluorocarbon attenuates response of concanavalin A-stimulated mononuclear blood cells without altering ligand-receptor interaction[J]. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1): L210-L216.
29.Fernandez R, Sarma V, Younkin E, et al. Exposure to perflubron is associated with decreased Syk phosphorylation in human neutrophils[J]. J Appl Physiol, 2001, 91(5): 1941-1947.
30.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
31.Rotta AT, Gunnarsson B, Fuhrman BP, et al. Perfluorooctyl bromide (perflubron) attenuates oxidative injury to biological and nonbiological systems[J]. Pediatr Crit Care Med, 2003, 4(2): 233-238.
32.Varani J, Hirschl RB, Dame M, et al. Perfluorocarbon protects lung epithelial cells fromneutrophil-mediated injury in an in vitro model of liquid ventilation therapy[J]. Shock, 1996, 6(5): 339-344.
33.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha- induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
34.Jung R, Pendland SL, Martin SJ. Effect of perfluorooctyl bromide on bacterial growth[J]. Chemotherapy, 2003, 49(1-2): 1-7.
2. Christman JW, Sadikot RT, Blackwell TS. The role of nuclear factor-κB in pulmonary diseases[J]. Chest, 2000, 117(5): 1482-1487.
3. Carden D, Xiao F, Moak C, et al. Neutrophil elastase promotes lung microvascular injury and proteolysis of endothelial cadherins[J]. Am J Physiol, 1998, 275(2 Pt 2): H385-392.
4. Wang Q, Yerukhimovich M, Gaarde WA, et al. MKK3 and -6-dependent activation of p38alpha MAP kinase is required for cytoskeletal changes in pulmonary microvascular endothelial cells induced by ICAM-1 ligation[J]. Am J Physiol Lung Cell Mol Physiol, 2005, 288(2): L359-369.
5. Hübler M, Souders JE, Shade ED, et al. Effects of vaporized perfluorocarbon on pulmonary blood flow and ventilation perfusion distribution in a model of acute respiratory distress syndrome[J]. Anesthesiology, 2001, 95(6): 1414-1421.
6. Bleyl JU, Ragaller M, Tsch? U, et al. Changes in pulmonary function and oxygenation during application of perfluorocarbon vapor in healthy and oleic acid-injured animals[J]. Crit Care Med, 2002, 30(6): 1340-1347.
7. van Eeden SF, Klut ME, Leal MA, et al. Partial liquid ventilation with perfluorocarbon in acute lung injury light and transmission electron microscopy studies[J]. Am J Respir Cell Mol Biol, 2000, 22(4): 441-450.
8. Colton DM, Till GO, Johnson KJ, et al. Partial liquid ventilation decreases albumin leak in the setting of acute lung injury[J]. J Crit Care, 1998, 13(3): 136-139.
9. Kawamae K, Pristine G, Chiumello D, et al. Partial liquid ventilation decreases serum tumor necrosis factor-alpha concentrations in a rat acid aspiration lung injury model[J]. Crit Care Med, 2000, 28(2): 479-483.
10.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary iInflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
11.Heard SO, Puyana JC. The anti-inflammatory effects of perfluorocarbons: let's get physical[J]. Crit Care Med, 2000, 28(4): 1241-1242.
12.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha- induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
13.Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
14.Cox C, Stavis RL, Wolfson MR, et al. Long-term tidal liquid ventilation in premature lambs: physiologic, biochemical and histological correlates[J]. Biol Neonate, 2003, 84(3): 232-242.
15.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
16.Rotta AT, Steinhorn DM. Partial liquid ventilation reduces pulmonary neutrophil accumulation in an experimental model of systemic endotoxemia and acute lung injury[J]. Crit Care Med, 1998, 26(10): 1707-1715.
17.Ellena JF, Obraztsov VV, Cumbea VL, et al. Perfluorooctyl bromide has limited membrane solubility and is located at the bilayer center. Locating small molecules in lipid bilayers through paramagnetic enhancements of NMR relaxation[J]. J Med Chem, 2002, 45(25): 5534-5542.
18.Nakstad B, Wolfson MR, Shaffer TH, et al. Perfluorochemical liquids modulate cell-mediated inflammatory responses[J]. Crit Care Med, 2001, 29(9): 1731-1737.
19.Fernandez R, Sarma V, Younkin E, et al. Exposure to perflubron is associated with decreased Syk phosphorylation in human neutrophils[J]. J Appl Physiol, 2001, 91(5): 1941-1947.
20.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
21.Colton DM, Till GO, Johnson KJ, et al. Neutrophil accumulation is reduced during partial liquid ventilation[J]. Crit Care Med, 1998, 26(10): 1716-1724.
22.Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
23.Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
24.Jeng MJ, Kou YR, Sheu CC, et al. Effects of partial liquid ventilation with FC-77 on acute lung injury in newborn piglets[J]. Pediatr Pulmonol, 2002, 33(1): 12-21.
25.Suh GY, Chung MP, Park SJ, et al. Partial liquid ventilation with perfluorocarbon improves gas exchange and decreases inflammatory response in oleic acid-induced lung injury in beagles[J]. J Korean Med Sci, 1999, 14(6): 613-622.
26.Croce MA, Fabian TC, Patton JH Jr, et al. Partial liquid ventilation decreases the inflammatory response in the alveolar environment of trauma patients[J]. J Trauma, 1998, 45(2): 273-280.
27.Rotta AT, Gunnarsson B, Hernan LJ, et al. Partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids[J]. Crit Care Med, 2000, 28(1): 202-208.
28.Dani C, Costantino ML, Martelli E, et al. Perfluorocarbons attenuate oxidative lung damage[J]. Pediatr Pulmonol, 2003, 36(4): 322-329.
29.Hirayama Y, Hirasawa H, Oda S, et al. Partial liquid ventilation with FC-77 suppresses the release of lipid mediators in rat acute lung injury model[J]. Crit Care Med, 2004, 32(10): 2085-2089.
30.Hirschl RB, Conrad S, Kaiser R, et al. Partial liquid ventilation in adult patients with ARDS: a multicenter phase I-II trial. Adult PLV Study Group[J]. Ann Surg, 1998, 228(5): 692-700.
31.Kandler MA, von der Hardt K, Schoof E, et al. Persistent improvement of gas exchange and lung mechanics by aerosolized perfluorocarbon[J]. Am J Respir Crit Care Med, 2001, 164(1): 31-35.
32.Ragaller M, Bleyl J, Tscho U, et al. Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury[J]. Intensive Care Med, 2001, 27(5): 889-897.
33.Thomas SR, Gradon L, Pratsinis SE, et al. Perfluorocarbon compound aerosols for delivery to the lung as potential 19F magnetic resonance reporters of regional pulmonary pO2[J]. Invest Radiol, 1997, 32(1): 29-38.
34.Hildebrand F, Pape HC, Harwood P, et al. Role of adhesion molecule ICAM in the pathogenesis of polymicrobial sepsis[J]. Exp Toxicol Pathol, 2005, 56(4-5): 281-290.
35.Reickert C, Pranikoff T, Overbeck M, et al. The pulmonary and systemic distribution and elimination of perflubron from adult patients treated with partial liquid ventilation[J]. Chest, 2001, 119(2): 515-522.
36.Varani J, Hirschl RB, Dame M, et al. Perfluorocarbon protects lung epithelial cells from neutrophil-mediated injury in an in vitro model of liquid ventilation therapy[J]. Shock, 1996, 6(5): 339-344.
37.Chi JT, Chang HY, Haraldsen G, et al. Endothelial cell diversity revealed by global expression profiling[J]. Proc Natl Acad Sci U S A, 2003, 100(19): 10623-10628.
38.King J, Hamil T, Creighton G, et al. Structural and functional characteristics of lung macro- and microvascular endothelial cell phenotypes[J]. Microvasc Res, 2004, 67(2): 139-151.
39.Medhora M, Daniels J, Mundey K, et al. Epoxygenase-driven angiogenesis in human lung microvascular endothelial cells[J]. Am J Physiol Heart Circ Physiol, 2003, 284(1): H215-H224.
40.Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J].Crit Care Med, 1997, 25(12): 2045-2047.
41.Koch T, Ragaller M, Haufe D, et al. Perfluorohexane attenuates proinflammatory and procoagulatory response of activated monocytes and alveolar macrophages[J]. Anesthesiology, 94(1): 101-109.
42.Smith TM, Steinhorn DM, Thusu K, et al. A liquid perfluorochemical decreases the in vitro production of reactive oxygen species by alveolar macrophages[J]. Crit Care Med, 1995, 23(9): 1533-1539.
43.Palsson-McDermott EM, O'Neill LA. Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4[J]. Immunology, 2004, 113(2): 153-162.
44.Poltorak A, He X, Smirnova I, et al. Defective LPS signaling in C3H/HeJ andC57BL/10ScCr mice: mutations in Tlr4 gene[J]. Science, 1998, 282(5396): 2085-2088.
45.Underhill DM, Ozinsky A. Toll-like receptors: key mediators of microbe detection[J]. Curr Opin Immunol, 2002, 14(1): 103-110.
46.Medzhitov R, Preston-Hurlburt P, Janeway CA Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J]. Nature, 1997, 388(6640): 394-397.
47.Nagai Y, Akashi S, Nagafuku M, et al. Essential role of MD-2 in LPS responsiveness and TLR4 distribution[J]. Nat Immunol, 2002, 3(7): 667-672.
48.Haufe D, Luther T, Kotzsch M, et al. Perfluorocarbon attenuates response of concanavalin A-stimulated mononuclear blood cells without altering ligand-receptor interaction[J]. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1): L210-L216.
49.Kobayashi T. Walsh MC. Choi Y. The role of TRAF6 in signal transduction and the immune response[J]. Microbes Infect, 2004, 6(14): 1333-1338.
50.Lomaga MA, Yeh WC, Sarosi I, et al. TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling[J]. Genes Dev, 1999, 13(8): 1015-1024.
1. Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
2. Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
3. Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
4. Clark LC, Gollan F. Survival of mammals breathing organic liquid sequilibrated with oxygenat at mospheric pressure[J].Science, 1966, 152(730): 1755-1756.
5. Fuhrman BP, Paczan PR, DeFrancisis M. Perfluorocarbon-associated gas exchange[J]. Crit Care Med, 1991, 19(5): 712-722.
6. Hirschl RB, Pranikoff T, Wise C, et al. Initial experience with partial liquid ventilation in adult patients with the acute respiratory distress syndrome[J]. JAMA, 1996, 275(5): 383-389.
7. Bleyl JU, Ragaller M, Tscho U, et al. Vaporized perfluorocarbon improves oxygenation and pulmonary function in an ovine model of acute respiratory distress syndrome[J]. Anesthesiology, 1999, 91(2): 461-469.
8. Thomas SR, Gradon L, Pratsinis SE, et al. Perfluorocarbon compound aerosols for delivery to the lung as potential 19F magnetic resonance reporters of regional pulmonary pO2[J]. Invest Radiol, 1997, 32(1): 29-38.
9. Thomas SH, O'Doherty MJ, Page CJ, et al. Delivery of ultrasonic nebulized aerosols to a lung model during mechanical ventilation. Am Rev Respir Dis, 1993, 148(4 Pt 1): 872-877.
10.Rudiger M, Gregor T, Burkhardt W, et al. Perfluorocarbon species and nebulizer type influence aerosolization rate and particle size of perfluorocarbon aerosol[J]. J Crit Care, 2004, 19(1): 42-47.
11.Gregor T, Schmalisch G, Burkhardt W, et al. Aerosolization of perfluorocarbons during mechanical ventilation: an in vitro study[J]. Intensive Care Med, 2003, 29(8): 1354-1360.
12.Ragaller M, Bleyl J, Tscho U, et al. Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury[J]. Intensive Care Med, 2001, 27(5): 889-897.
13.Kandler MA, von der Hardt K, Schoof E, et al. Persistent improvement of gas exchange and lung mechanics by aerosolized perfluorocarbon[J]. Am J Respir Crit Care Med, 2001, 164(1): 31-35.
14.Bleyl JU, Ragaller M, Tsch? U, et al. Changes in pulmonary function and oxygenation during application of perfluorocarbon vapor in healthy and oleic acid-injured animals[J]. Crit Care Med, 2002, 30(6): 1340-1347.
15.Hübler M, Souders JE, Shade ED, et al. Effects of vaporized perfluorocarbon on pulmonary blood flow and ventilation perfusion distribution in a model of acute respiratory distress syndrome[J]. Anesthesiology, 2001, 95(6): 1414-1421.
16.Hubler M, Heller AR, Bleyl JU, et al. Perfluorohexane vapor has only minor effects on spatial pulmonary blood flow distribution in isolated rabbit lungs[J]. Anesth Analg, 2005, 100(4): 1122-1128.
17.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
18.Schoof E, von der Hardt K, Kandler MA, et al. Aerosolized perfluorocarbon reduces adhesion molecule gene expression and neutrophil sequestration in acute respiratory distress[J]. Eur J Pharmacol, 2002, 457(2-3): 195-200.
19.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
20.Gama de Abreu M, Wilmink B, Hubler M, et al. Vaporized perfluorohexane attenuates ventilator-induced lung injury in isolated, perfused rabbit lungs[J]. Anesthesiology, 2005, 102(3): 597-605.
21.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha-induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
22.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
1. Obraztsov VV, Neslund GG, Kombrust ES, et al. In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J]. Am J Physiol Lung Cell Mol Physiol, 2000, 278(5): L1018-L1024.
2. Nakstad B, Wolfson MR, Shaffer TH, et al. Perfluorochemical liquids modulate cell-mediated inflammatory responses[J]. Crit Care Med, 2001, 29(9): 1731-1737.
3. Shaffer TH, Wolfson MR, Greenspan JS, et al. Liquid ventilation in premature lambs: uptake, biodistribution and elimination of perfluorodecalin liquid[J]. Reprod Fertil Dev, 1996, 8(3): 409-416.
4. Cox C, Stavis RL, Wolfson MR, et al. Long-term tidal liquid ventilation in premature lambs: physiologic, biochemical and histological correlates[J]. Biol Neonate, 2003, 84(3): 232-242.
5. Reickert C, Pranikoff T, Overbeck M, et al. The pulmonary and systemic distribution and elimination of perflubron from adult patients treated with partial liquid ventilation[J]. Chest, 2001, 119(2): 515-522.
6. Ellena JF, Obraztsov VV, Cumbea VL, et al. Perfluorooctyl bromide has limited membrane solubility and is located at the bilayer center. Locating small molecules in lipid bilayers through paramagnetic enhancements of NMR relaxation[J]. J Med Chem, 2002, 45(25): 5534-5542.
7. Rüdiger M, Wissel H, Ochs M, et al. Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J]. Crit Care Med, 2003, 31(4): 1190-1196.
8. Koch T, Ragaller M, Haufe D, et al. Perfluorohexane attenuates proinflammatory and procoagulatory response of activated monocytes and alveolar macrophages[J]. Anesthesiology, 94(1): 101-109.
9. Quintel M, Heine M, Hirschl RB, et al. Effects of partial liquid ventilation on lung injury in a model of acute respiratory failure: a histologic and morphometric analysis[J]. Crit Care Med, 1998, 26(5): 833-843.
10.van Eeden SF, Klut ME, Leal MA, et al. Partial liquid ventilation with perfluorocarbonin acute lung injury light and transmission electron microscopy studies[J]. Am J Respir Cell Mol Biol, 2000, 22(4): 441-450.
11.Colton DM, Till GO, Johnson KJ, et al. Partial liquid ventilation decreases albumin leak in the setting of acute lung injury[J]. J Crit Care, 1998, 13(3): 136-139.
12.Lange NR, Kozlowski JK, Gust R, et al. Effect of partial liquid ventilation on pulmonary vascular permeability and edema after experimental acute lung injury[J]. Am J Respir Crit Care Med, 2000, 162(1): 271-277.
13.Rotta AT, Steinhorn DM. Partial liquid ventilation reduces pulmonary neutrophil accumulation in an experimental model of systemic endotoxemia and acute lung injury[J]. Crit Care Med, 1998, 26(10): 1707-1715.
14.Colton DM, Till GO, Johnson KJ, et al. Neutrophil accumulation is reduced during partial liquid ventilation[J]. Crit Care Med, 1998, 26(10): 1716-1724.
15.Nader ND, Knight PR, Davidson BA, et al. Systemic perfluorocarbons suppress the acute lung inflammation after gastric acid aspiration in rats[J]. Anesth Analg, 2000, 90(2): 356-361.
16.Suh GY, Chung MP, Park SJ, et al. Partial liquid ventilation with perfluorocarbon improves gas exchange and decreases inflammatory response in oleic acid-induced lung injury in beagles[J]. J Korean Med Sci, 1999, 14(6): 613-622.
17.Steinhorn DM, Papo MC, Rotta AT, et al. Liquid ventilation attenuates pulmonary oxidative damage[J]. J Crit Care, 1999, 14(1): 20-28.
18.Rotta AT, Gunnarsson B, Hernan LJ, et al. Partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids[J]. Crit Care Med, 2000, 28(1): 202-208.
19.Dani C, Costantino ML, Martelli E, et al. Perfluorocarbons attenuate oxidative lung damage[J]. Pediatr Pulmonol, 2003, 36(4): 322-329.
20.Hirayama Y, Hirasawa H, Oda S, et al. Partial liquid ventilation with FC-77 suppresses the release of lipid mediators in rat acute lung injury model[J]. Crit Care Med, 2004, 32(10): 2085-2089.
21.Schoof E, von der Hardt K, Kandler MA, et al. Aerosolized perfluorocarbon reduces adhesion molecule gene expression and neutrophil sequestration in acute respiratory distress[J]. Eur J Pharmacol, 2002, 457(2-3):195-200.
22.von der Hardt K, Kandler MA, Brenn G, et al. Comparison of aerosol therapy with different perfluorocarbons in surfactant-depleted animals[J]. Crit Care Med, 2004, 32(5): 1200-1206.
23.Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J].Crit Care Med, 1997, 25(12): 2045-2047.
24.Kawamae K, Pristine G, Chiumello D, et al. Partial liquid ventilation decreases serum tumor necrosis factor-alpha concentrations in a rat acid aspiration lung injury model[J]. Crit Care Med, 2000, 28(2): 479-483.
25.von der Hardt K, Schoof E, Kandler MA, et al. Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model[J]. Pediatr Res, 2002, 51(2): 177-182.
26.von der Hardt K, Kandler MA, Fink L, et al. Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon[J]. Am J Physiol Lung Cell Mol Physiol, 2003, 285(1): L55-L62.
27.Croce MA, Fabian TC, Patton JH Jr, et al. Partial liquid ventilation decreases the inflammatory response in the alveolar environment of trauma patients[J]. J Trauma, 1998, 45(2): 273-280.
28.Haufe D, Luther T, Kotzsch M, et al. Perfluorocarbon attenuates response of concanavalin A-stimulated mononuclear blood cells without altering ligand-receptor interaction[J]. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1): L210-L216.
29.Fernandez R, Sarma V, Younkin E, et al. Exposure to perflubron is associated with decreased Syk phosphorylation in human neutrophils[J]. J Appl Physiol, 2001, 91(5): 1941-1947.
30.Chang H, Kuo FC, Lai YS, et al. Inhibition of inflammatory responses by FC-77, a perfluorochemical, in lipopolysaccharide-treated RAW 264.7 macrophages[J]. Intensive Care Med, 2005, 31(7): 977-984.
31.Rotta AT, Gunnarsson B, Fuhrman BP, et al. Perfluorooctyl bromide (perflubron) attenuates oxidative injury to biological and nonbiological systems[J]. Pediatr Crit Care Med, 2003, 4(2): 233-238.
32.Varani J, Hirschl RB, Dame M, et al. Perfluorocarbon protects lung epithelial cells fromneutrophil-mediated injury in an in vitro model of liquid ventilation therapy[J]. Shock, 1996, 6(5): 339-344.
33.Baba A, Kim YK, Zhang H, et al. Perfluorocarbon blocks tumor necrosis factor-alpha- induced interleukin-8 release from alveolar epithelial cells in vitro[J]. Crit Care Med, 2000, 28(4): 1113-1118.
34.Jung R, Pendland SL, Martin SJ. Effect of perfluorooctyl bromide on bacterial growth[J]. Chemotherapy, 2003, 49(1-2): 1-7.