全氟化碳对A549细胞钠离子通道表达影响的研究
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摘要
目的:
探讨全氟化碳原液(PFC)和新型全氟化碳乳剂(NPE)对A549细胞生长的影响;观察PFC和NPE对TNF-α攻击的A549细胞钠离子通道(ENaC)表达的影响。
方法:
1.PFC对A549细胞生长影响及对TNF-α攻击的A549细胞ENaC表达影响的研究。分为对照组、TNF-α组、PFC组和治疗组共四组。TNF-α以100ng/ml的浓度攻击细胞,PFC以1/10细胞培养液体积的比例与细胞共同培养。正常组细胞不做任何处理,实验组细胞分别用不同因素处理2、4、6h,用MTT法测各组细胞活力变化;提取细胞RNA和蛋白,分别用real-time PCR法检测细胞ENaCα、β、γ亚单位表达水平变化;ENaC-α蛋白表达水平采用Western-blot法检测。
2. NPE对A549细胞生长影响及对TNF-α攻击的A549细胞ENaC表达影响的研究。分为对照组、TNF-α组、NPE组和治疗组共四组。TNF-α以100ng/ml的浓度攻击细胞,NPE以1/10细胞培养液体积的比例与细胞共同培养。正常组细胞不做任何处理,实验组细胞分别用不同因素处理2、4、6h,用MTT法测各组细胞活力变化;提取细胞RNA和蛋白,分别用real-time PCR法检测ENaCα、β、γ亚单位表达水平变化;ENaC-α蛋白表达水平采用Western-blot法检测。
结果:
1. PFC对A549细胞生长影响:2h组:与对照组细胞相比较:TNF-α组、PFC组和治疗组细胞基本形态无明显变化。4h组:TNF-α组细胞细胞生长状态不良,死亡细胞较多。PFC组和治疗组细胞一般情况较TNF-α组好,死亡细胞数较少。6h各组细胞一般情况与4h组无明显差别。
2. PFC对TNF-α攻击的A549细胞活力的影响:2h组:TNF-α组、PFC组、治疗组细胞吸光度值与对照组比较,差异均无统计学意义(P﹥0.05)。4h组:PFC组细胞吸光度值为与对照组比较,差异无统计学意义(P﹥0.05);TNF-α组、治疗组细胞吸光度值较对照组降低,治疗组吸光度值较TNF-α组增高,差异具有统计学意义(P﹤0.05)。6h各组吸光度值比较结果与4h组相似。
3. real-time PCR结果显示:2h组:与对照组相比,各组ENaC-α、β、γ亚单位mRNA水平变化不明显;处理4h后,与对照组相比,TNF-α组细胞ENaC各亚单位mRNA水平均有不同程度的降低,PFC组细胞则与对照组无明显差异,治疗组细胞ENaC各亚单位mRNA水平也有不同程度降低,与TNF-α组同时相点相比,差异无统计学意义。6h组情况与4h组相同。
4. Western-blot结果显示:2h组:与对照组相比,各组ENaC-α蛋白表达变化不明显;4h组和6h组中,有TNF-α组细胞ENaC-α蛋白表达明显下调,其他各组细胞ENaC-α蛋白表达与对照组并无差别。
5.NPE对A549细胞生长影响:2h组:与对照组细胞相比较:TNF-α组、NPE组、治疗组细胞基本形态无明显变化。4h组:TNF-α致伤组细胞细胞生长状态不良,死亡细胞较多。NPE组和治疗组细胞一般情况较TNF-α组好,细胞形态变化不明显,死亡细胞数较少。6h组各组细胞一般情况与4h组无明显区别。
6. NPE对TNF-α攻击的A549细胞活力的影响: 2h组:NPE组、TNF-α组、治疗组细胞吸光度值与对照组比较,无明显差异(P﹥0.05)。4h组:NPE组细胞吸光度值与对照组比较,无明显差异(P﹥0.05);TNF-α组、治疗组细胞吸光度值较对照组明显降低,治疗组吸光度值较T组增高,差异具有统计学意义(P﹤0.05)。6h各组吸光度值比较结果与4h组相似。
7. real-time PCR结果显示:2h组:与对照组相比,各组ENaC-α、β、γ亚单位mRNA水平变化不明显;处理4h后,与对照组相比,TNF-α组细胞ENaC各亚单位mRNA水平均有不同程度的降低,NPE组细胞则与对照组无明显差别,治疗组细胞ENaC各亚单位mRNA水平也有不同程度降低,但较TNF-α组同时相点相比有明显升高,差异具有统计学意义。6h组情况与4h组相同。
8. Western-blot结果显示:2h组:与对照组相比,各组ENaC-α蛋白表达变化不明显;4h组和6h组中, TNF-α组细胞ENaC-α蛋白表达明显下调,治疗组细胞ENaC-α蛋白表达与TNF-α组相比有明显增高,差异具有统计学意义(P﹤0.05),NPE组细胞ENaC-α蛋白表达与对照组无明显差异。
结论:
1. TNF-α可对A549细胞生长及活力产生抑制作用,且可以下调细胞ENaC-α、β、γ亚单位及ENaC-α蛋白表达水平。
2.PFC可能通过物理屏障作用对TNF-α攻击的A549细胞产生一定的保护作用,对细胞ENaC-α、β、γ亚单位及ENaC-α蛋白表达的下调无改善作用。
3.NPE可改善TNF-α对A549细胞生长的抑制作用,同时可以改善TNF-α对细胞ENaC各基因及ENaC-α蛋白表达的下调。
Objective:
To explore the impaction of perfluorocarbon and new perfluorocarbon emulsion on A549 cells; To investigate the protection and mechanism of PFC and NPE on TNF-αattacked A549 cells.
Methods:
1. The exploration of PFC: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand PFC group with PFC at a 1/10 volume; we used the same dose of TNF-αand PFC in treat group. After treatment for 2、4、6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α、β、γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively.
2. The exploration of NPE: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand NPE group with NPE at a 1/10 volume; we used the same dose of TNF-αand NPE in treat group. After treatment for 2、4、6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α、β、γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively.
Results:
1. Compared with control group, the cell morphology of other groups had no significant change after treated for 2 hours; however, when it came to 4 and 6 hours, the cells in TNF-αgroup grew alas, and many cells died and float. Meanwhile, the cells in PFC group and treat group had no difference with control group (P﹥0.05).
2. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).
3.The mRNA levels of ENaC-α、β、γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and there was no difference between TNF-αgroup and treat group(P﹥0.05). There was no significant change in PFC group compared with control group after treated for 4 and 6 hours( P﹥0.05).
4. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, there was no difference between treat group and TNF-αgroup(P﹥0.05). The mRNA levels of PFC group had no difference with group C(P﹥0.05).
5. The cell morphology of each group had no significant difference after treated for 2 hours; however, when incubated for 4 and 6 hours, the cells in TNF-αgroup grew bad, and many cells died and float. Meanwhile, the cells in NPE group and treat group had no difference with control group (P﹥0.05).
6. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).
7. The mRNA levels of ENaC-α、β、γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and the treat group had a higer level than TNF-αgroup (P﹤0.05). There was no significant change in NPE group compared with control group after treated for 4 and 6 hours( P﹥0.05).
8. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, and the level of treat group was higer than TNF-αgroup (P﹤0.05). But the NPE group has the same level as control group (P﹥0.05).
Conclusion:
1. Incubated with TNF-αimpacted the morphology and cell activity of A549 cell.When the incubation last to 4 and 6 hours, the mRNA expression of ENaC-α、β、γsubunits and protein level of ENaC-αwere decreased.
2. PFC protected A549 cells attacted with TNF-αby physical barriers, but has no ameliorations in the mRNA and protein expression.
3. NPE released the depression of TNF-α, and aliveiated the downregulation of TNF-αto the mRNA and protein expression of ENaC in A549 cells.
探讨全氟化碳原液(PFC)和新型全氟化碳乳剂(NPE)对A549细胞生长的影响;观察PFC和NPE对TNF-α攻击的A549细胞钠离子通道(ENaC)表达的影响。
方法:
1.PFC对A549细胞生长影响及对TNF-α攻击的A549细胞ENaC表达影响的研究。分为对照组、TNF-α组、PFC组和治疗组共四组。TNF-α以100ng/ml的浓度攻击细胞,PFC以1/10细胞培养液体积的比例与细胞共同培养。正常组细胞不做任何处理,实验组细胞分别用不同因素处理2、4、6h,用MTT法测各组细胞活力变化;提取细胞RNA和蛋白,分别用real-time PCR法检测细胞ENaCα、β、γ亚单位表达水平变化;ENaC-α蛋白表达水平采用Western-blot法检测。
2. NPE对A549细胞生长影响及对TNF-α攻击的A549细胞ENaC表达影响的研究。分为对照组、TNF-α组、NPE组和治疗组共四组。TNF-α以100ng/ml的浓度攻击细胞,NPE以1/10细胞培养液体积的比例与细胞共同培养。正常组细胞不做任何处理,实验组细胞分别用不同因素处理2、4、6h,用MTT法测各组细胞活力变化;提取细胞RNA和蛋白,分别用real-time PCR法检测ENaCα、β、γ亚单位表达水平变化;ENaC-α蛋白表达水平采用Western-blot法检测。
结果:
1. PFC对A549细胞生长影响:2h组:与对照组细胞相比较:TNF-α组、PFC组和治疗组细胞基本形态无明显变化。4h组:TNF-α组细胞细胞生长状态不良,死亡细胞较多。PFC组和治疗组细胞一般情况较TNF-α组好,死亡细胞数较少。6h各组细胞一般情况与4h组无明显差别。
2. PFC对TNF-α攻击的A549细胞活力的影响:2h组:TNF-α组、PFC组、治疗组细胞吸光度值与对照组比较,差异均无统计学意义(P﹥0.05)。4h组:PFC组细胞吸光度值为与对照组比较,差异无统计学意义(P﹥0.05);TNF-α组、治疗组细胞吸光度值较对照组降低,治疗组吸光度值较TNF-α组增高,差异具有统计学意义(P﹤0.05)。6h各组吸光度值比较结果与4h组相似。
3. real-time PCR结果显示:2h组:与对照组相比,各组ENaC-α、β、γ亚单位mRNA水平变化不明显;处理4h后,与对照组相比,TNF-α组细胞ENaC各亚单位mRNA水平均有不同程度的降低,PFC组细胞则与对照组无明显差异,治疗组细胞ENaC各亚单位mRNA水平也有不同程度降低,与TNF-α组同时相点相比,差异无统计学意义。6h组情况与4h组相同。
4. Western-blot结果显示:2h组:与对照组相比,各组ENaC-α蛋白表达变化不明显;4h组和6h组中,有TNF-α组细胞ENaC-α蛋白表达明显下调,其他各组细胞ENaC-α蛋白表达与对照组并无差别。
5.NPE对A549细胞生长影响:2h组:与对照组细胞相比较:TNF-α组、NPE组、治疗组细胞基本形态无明显变化。4h组:TNF-α致伤组细胞细胞生长状态不良,死亡细胞较多。NPE组和治疗组细胞一般情况较TNF-α组好,细胞形态变化不明显,死亡细胞数较少。6h组各组细胞一般情况与4h组无明显区别。
6. NPE对TNF-α攻击的A549细胞活力的影响: 2h组:NPE组、TNF-α组、治疗组细胞吸光度值与对照组比较,无明显差异(P﹥0.05)。4h组:NPE组细胞吸光度值与对照组比较,无明显差异(P﹥0.05);TNF-α组、治疗组细胞吸光度值较对照组明显降低,治疗组吸光度值较T组增高,差异具有统计学意义(P﹤0.05)。6h各组吸光度值比较结果与4h组相似。
7. real-time PCR结果显示:2h组:与对照组相比,各组ENaC-α、β、γ亚单位mRNA水平变化不明显;处理4h后,与对照组相比,TNF-α组细胞ENaC各亚单位mRNA水平均有不同程度的降低,NPE组细胞则与对照组无明显差别,治疗组细胞ENaC各亚单位mRNA水平也有不同程度降低,但较TNF-α组同时相点相比有明显升高,差异具有统计学意义。6h组情况与4h组相同。
8. Western-blot结果显示:2h组:与对照组相比,各组ENaC-α蛋白表达变化不明显;4h组和6h组中, TNF-α组细胞ENaC-α蛋白表达明显下调,治疗组细胞ENaC-α蛋白表达与TNF-α组相比有明显增高,差异具有统计学意义(P﹤0.05),NPE组细胞ENaC-α蛋白表达与对照组无明显差异。
结论:
1. TNF-α可对A549细胞生长及活力产生抑制作用,且可以下调细胞ENaC-α、β、γ亚单位及ENaC-α蛋白表达水平。
2.PFC可能通过物理屏障作用对TNF-α攻击的A549细胞产生一定的保护作用,对细胞ENaC-α、β、γ亚单位及ENaC-α蛋白表达的下调无改善作用。
3.NPE可改善TNF-α对A549细胞生长的抑制作用,同时可以改善TNF-α对细胞ENaC各基因及ENaC-α蛋白表达的下调。
Objective:
To explore the impaction of perfluorocarbon and new perfluorocarbon emulsion on A549 cells; To investigate the protection and mechanism of PFC and NPE on TNF-αattacked A549 cells.
Methods:
1. The exploration of PFC: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand PFC group with PFC at a 1/10 volume; we used the same dose of TNF-αand PFC in treat group. After treatment for 2、4、6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α、β、γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively.
2. The exploration of NPE: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand NPE group with NPE at a 1/10 volume; we used the same dose of TNF-αand NPE in treat group. After treatment for 2、4、6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α、β、γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively.
Results:
1. Compared with control group, the cell morphology of other groups had no significant change after treated for 2 hours; however, when it came to 4 and 6 hours, the cells in TNF-αgroup grew alas, and many cells died and float. Meanwhile, the cells in PFC group and treat group had no difference with control group (P﹥0.05).
2. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).
3.The mRNA levels of ENaC-α、β、γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and there was no difference between TNF-αgroup and treat group(P﹥0.05). There was no significant change in PFC group compared with control group after treated for 4 and 6 hours( P﹥0.05).
4. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, there was no difference between treat group and TNF-αgroup(P﹥0.05). The mRNA levels of PFC group had no difference with group C(P﹥0.05).
5. The cell morphology of each group had no significant difference after treated for 2 hours; however, when incubated for 4 and 6 hours, the cells in TNF-αgroup grew bad, and many cells died and float. Meanwhile, the cells in NPE group and treat group had no difference with control group (P﹥0.05).
6. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).
7. The mRNA levels of ENaC-α、β、γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and the treat group had a higer level than TNF-αgroup (P﹤0.05). There was no significant change in NPE group compared with control group after treated for 4 and 6 hours( P﹥0.05).
8. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, and the level of treat group was higer than TNF-αgroup (P﹤0.05). But the NPE group has the same level as control group (P﹥0.05).
Conclusion:
1. Incubated with TNF-αimpacted the morphology and cell activity of A549 cell.When the incubation last to 4 and 6 hours, the mRNA expression of ENaC-α、β、γsubunits and protein level of ENaC-αwere decreased.
2. PFC protected A549 cells attacted with TNF-αby physical barriers, but has no ameliorations in the mRNA and protein expression.
3. NPE released the depression of TNF-α, and aliveiated the downregulation of TNF-αto the mRNA and protein expression of ENaC in A549 cells.
引文
1 樊毫军,刘书盈,张健鹏,等.静脉注射内毒素致大鼠急性肺损伤模型的病理生理学指标评价。中国危重病急救医学,2006,18(8):485-487;
2 Matthay MA , Clerici C , Saumon G,et al. Active fluid clearance from the distal air spaces of the lung . J Appl Physiol 2002, 93: 1533-1541.
3 Berthiaume Y, Folkesson HG, Matthay MA,et al. Lung edema clearance: 20 years of progress: invited review: alveolar edema fluid clearance in the injued lung. Am J Physiol, 2002, 93(6):2207-2213.
4 Norlin A, Lu LN,Guggino SE ,et al. Contribution of amiloride insensitive pathways to alveolar fluid clearance in adult rats. J Appl Physiol, 2001,90(4):1489-1496.
5 Fiers W, Jayr C, Lazrak A, et,al.Tumor necrosis factor. Characterization at the molecular,cellular and in vivo level. FEBS Lett 1999;285:199-212
6 Steinhorn DM, Leach CL, Fuhrman BP, et al. Partial liquid ventilation enhances surfactant phospholipids production. Crit Care Med,1996,24(7):1252-1256.
7 Fukuda N, Jayr C, Lazrak A, et,al. Mechanisms of TNF-α stimulation of amiloride-sensitivesodium transport across alveolar epithelium. Am J Physiol Lung Cell MolPhysiol 280: L1258–L1265, 2001.
8 Nakamura H,Yoshimura K, Bajocchi G, et al. Tumor necrosis factor modulation of expression of thecystic fibrosistransmembrane conductance regulator gene. FEBS Lett 314:366–370, 1992.
9 Kellenberger S,Hoffmann Pochon N,Gautschi H, et al.On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
10 崔俊昌,刘又宁.全氟化碳的医学应用.北京医学,2001,23(3):168-170.
11 Nakamura H, Yoshimura K, Bajocchi G, et al. Tumor necrisis factor modulation of expression of the cystic fibrosis trasmembrane conductance regulator gene. FEBS Lett 1992;314:366-370.
12 张秋金,李银平,李檀实. 肺泡上皮细胞功能特性与内毒素性急性肺损伤.中国危重病急救医学,2005,17(6):382-384.
13 Woods CM,Gerald N,Elisabeth K,et al.Perflubron attenuates neutrophil adhesion to activated endothelial cells in vitro.Am J Physiol Lung Cell Mol Physiol,2000,278(5):L1008–L1017.
14 A Lazrak, Samanta S. Matalon.Biophysical properties and molecular characterization of amiloride-sensitive solium channels in A549 cells.Am J Physiol Lung Cell Mol Physiol.2000;278:L848-L857.
15 Andre Dagenais, Rosalie Frechette, Yuko Yamagata. Downregulation of ENaC activity and expression by TNF-αin alveolar epithelial cells. Am J Physiol Lung Cell Mol Physiol 2004;286: L301–L311.
16 樊毫军,刘书盈,张健鹏.全氟化碳对肿瘤坏死因子攻击下肺泡上皮细胞产生 IL-8 水平的影响.中华医学杂志,2006;86(15):1075-1076。
17 Alvarez RD, Canessa CM, Fyfe, et al. Structure and regulation of amiloride-sensitive sodium channel . Annu Rev Physiol, 2000, 62: 573-594.
18 Kellenberger S,Hoffmann Pochon N, Gautschi H, et al .On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
19 Carr S, Cantor JP, Cowan S,et al. Utilizing the peritoneum for gas exchange with oxygenated perfluorocarbons. American College of Surgeons 90th Annual Cliniacl Congress, New Orleans,2004.
20 Carr S, Cantor JP, Rao AS, et al. Peritoneal perfusion with Oxygenated Perfluorocarbon Augments Systemic Oxygenation. Chest,2006,130:402-411.
21 王晓光,赵晓巍,张健鹏. 全氟化碳雾化吸入对急性呼吸窘迫综合症猪气体交换、呼吸力学和血流动力学的影响.中华结核和呼吸杂志 2006;29(2):104-108.
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 M,Buhrow L,Wiedemann H.Perflubron decreases cytokine production by human alveolar macrophages.Crit Care Med,1997,25:2045–2047.
24 Woods CM,Gerald N,Elisabeth K,et al.Perflubron attenuates neutrophil adhesion to activated endothelial cells in vitro.Am J Physiol Lung Cell Mol Physiol,2000,278(5):L1008–L1017.
25 A Lazrak, Samanta K,Venetsanou, et al. Modification of biophysical properties of lung epithelial Na+ channels by dexamethasone. Am J Physiol 2000,279:762-770.
26 Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J]. Crit Care Med,1997,25(12):2045-2047.
27 马利波,刘文. 全氟化碳的临床应用及其并发症. 眼科,2003,12(3):182-186.
28 刘书盈,樊毫军,张健鹏. 全身应用全氟化碳防护内毒素致急性肺损伤可行性研究. 山东医药,2006;46(34):18-19.
29 樊毫军,刘书盈,张健鹏.全氟化碳腹腔注射对内毒素肺损伤大鼠中性粒细胞肺侵润的抑制作用.中华结核与呼吸杂志 2006;29(10):674-678.
1 樊毫军,刘书盈,张健鹏,等.静脉注射内毒素致大鼠急性肺损伤模型的病理生理学指标评价。中国危重病急救医学,2006,18(8):485-487;
2 Matthay MA , Clerici C , Saumon G, et al. Active fluid clearance from the distal air spaces of the lung . J Appl Physiol 2002, 93: 1533-1541.
3 Berthiaume Y, Folkesson HG, Matthay MA,et al. Lung edema clearance: 20 years of progress: invited review: alveolar edema fluid clearance in the injued lung. Am J Physiol, 2002, 93(6):2207-2213.
4 Norlin A, Lu LN,Guggino SE ,et al. Contribution of amiloride insensitive pathways to alveolar fluid clearance in adult rats. J Appl Physiol, 2001,90(4):1489-1496.
5 张秋金,李银平,李檀实.肺泡上皮细胞功能特性与内毒素性急性肺损伤.中国危重病急救医学,2005,17(6):382-384.
6 Alvarez RD, Canessa CM, Fyfe GK, et al ,Structure and regulation of amiloride-sensitive sodium channel . Annu Rev Physiol, 2000, 62: 573-594.
7 Kellenberger S,Hoffmann Pochon N, Gautschi H, et al ,On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
8 Richard, Ramminger , Inglis, et al. O2 can raise fetal pneumocyte Na + conductance without affecting-ENaC-mRNAabundance.Biochem-Biophys-Res-Commun. 2003, 305(3): 671-676;
9 Matthay MA, Folkesson HG, Clerici C. Lung epithelial fluid transport and the resolution of pulmonary edema. Phsiol Rev 2002, 82(3):569-600;
10 Mairbaurl H, Mayer K, Kim KJ, et al. Hypoxia decreases active Na+ transport across primary rat alveolar epithelial cell monolayers. Am J Physiol Lung Cell Mol Physiol, 2002, 282(4):L659-665;
11 Chen XJ,Eaton DC,Jain L.Beta-adrenergic regulation of amiloride-sensitive lung sodium channels.Am J Physiol Lung Cell Mol Physiol,2002,282:L609-L620.
12 Dobbs LG, Gonzalez R , Matthay MA, et al. Highly water permeable type Ⅰ alveolar epithelial cells confer high water permeability between the airspace and vasculature in rat lung. Proc Nalt Acad Sci USA ,1998,95:2991-2996.
13 Matalon S, Lazrak A, Lucky J,et al. Lung clearance 20years lf progress inveted review :Biophysical properties of sodem channels in lung alveolar epithelial cells.J Appl Physiol , 2002, 93:1852-1859;
14 Mustafa SB, Geronimo RJ, et al. postatal glucocorticoids induce alpha-ENaCformation and regulate glucocorticoids receptors in the pretern rabbit lung. Am Physiol Lung Cell Mol physiol,2004, 286(1):L73-80.
15 宋志芳,谢伟,单慧敏,等.糖皮质激素治疗大肠杆菌致急性呼吸窘迫综合症的实验研究,中国危重病急救医学,2006,18(12):716-720.
16 Suzuki S, Matsuda Y, Sugawara T, et al.Effects of carbenoxolone on alveolar fluid clearance and lung inflammation in the rat. Crit Care Med, 2004, 32: 1910-1915.
17 Caldwell,Boucher, Stutts M.J. Neutrophil elastase activates near-silent epithelial Na+ channels and increases airway epithelial Na + transport. Am.J.Physiol Lung Cell Mol.Physiol.2005,288:813-819.
18 Matthay MA, Fukuda N, Frank J, et al. Alveolar epithelial barrier: role in lung fluid balance in clinical lung injury.Clinic Chest Med, 2000,21:477-490.
19 Benos DJ, Awayda MS,Ismailov, et al. Structure and function of amiloride-sensitive epithelial Na+ channels. J Membrane Biol, 1995,143:1-18.
20 McRitchie DI, Isowa N, Edelson JD, et al. Production of tumour necrosis factor by primary cultured rat alveolarepithelial cells. Cytokine, 2000,12: 644–654.
21 Suter PM, Suter S, Girardin E, et al. High bronchoalveolar levels of tumor necrosis factor and its inhibitors, interleukin-1, interferon, and elastase, in patients with adult respiratory distress syndrome after trauma, shock, or sepsis. Am Rev Respir Dis, 1992 145: 1016–1022.
22 Dagenais A,Frechette R,Yamagata Y,et al.Downregulation of EnaC activity and expression by TNF-alpha in alveolar epithelial cells.Am J Physiol Lung Cell Mol Physiol,2004,286:L301-L311.
23 Frank J,Roux J,Kawakatsu H,et al.Transforming growth factor –beta1 decreases expression of the epithelial sodium channel alphaENaC and alveolar epithelial vectorial sodium and fluid transport via ERK1/2-dependentmechnism.J Biol Chem,2003,278(45):43939-43950.
24 Towne JE,Harrod KS, Krane CM ,et al. Decreased expression of aquaporin (AQP)1 and AQP5 in mouse lung after acute viral infection. Am J Respir Cell Mol Biol, 2000, 22: 34-44.
25 Dagenais A, Gosselin D, Guilbault C, et al. Modulation of epithelial sodium channel expression in mouse lung infected with Pseudomonas aeruginosa. Respir. Res, 2005,6(6): 2.
2 Matthay MA , Clerici C , Saumon G,et al. Active fluid clearance from the distal air spaces of the lung . J Appl Physiol 2002, 93: 1533-1541.
3 Berthiaume Y, Folkesson HG, Matthay MA,et al. Lung edema clearance: 20 years of progress: invited review: alveolar edema fluid clearance in the injued lung. Am J Physiol, 2002, 93(6):2207-2213.
4 Norlin A, Lu LN,Guggino SE ,et al. Contribution of amiloride insensitive pathways to alveolar fluid clearance in adult rats. J Appl Physiol, 2001,90(4):1489-1496.
5 Fiers W, Jayr C, Lazrak A, et,al.Tumor necrosis factor. Characterization at the molecular,cellular and in vivo level. FEBS Lett 1999;285:199-212
6 Steinhorn DM, Leach CL, Fuhrman BP, et al. Partial liquid ventilation enhances surfactant phospholipids production. Crit Care Med,1996,24(7):1252-1256.
7 Fukuda N, Jayr C, Lazrak A, et,al. Mechanisms of TNF-α stimulation of amiloride-sensitivesodium transport across alveolar epithelium. Am J Physiol Lung Cell MolPhysiol 280: L1258–L1265, 2001.
8 Nakamura H,Yoshimura K, Bajocchi G, et al. Tumor necrosis factor modulation of expression of thecystic fibrosistransmembrane conductance regulator gene. FEBS Lett 314:366–370, 1992.
9 Kellenberger S,Hoffmann Pochon N,Gautschi H, et al.On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
10 崔俊昌,刘又宁.全氟化碳的医学应用.北京医学,2001,23(3):168-170.
11 Nakamura H, Yoshimura K, Bajocchi G, et al. Tumor necrisis factor modulation of expression of the cystic fibrosis trasmembrane conductance regulator gene. FEBS Lett 1992;314:366-370.
12 张秋金,李银平,李檀实. 肺泡上皮细胞功能特性与内毒素性急性肺损伤.中国危重病急救医学,2005,17(6):382-384.
13 Woods CM,Gerald N,Elisabeth K,et al.Perflubron attenuates neutrophil adhesion to activated endothelial cells in vitro.Am J Physiol Lung Cell Mol Physiol,2000,278(5):L1008–L1017.
14 A Lazrak, Samanta S. Matalon.Biophysical properties and molecular characterization of amiloride-sensitive solium channels in A549 cells.Am J Physiol Lung Cell Mol Physiol.2000;278:L848-L857.
15 Andre Dagenais, Rosalie Frechette, Yuko Yamagata. Downregulation of ENaC activity and expression by TNF-αin alveolar epithelial cells. Am J Physiol Lung Cell Mol Physiol 2004;286: L301–L311.
16 樊毫军,刘书盈,张健鹏.全氟化碳对肿瘤坏死因子攻击下肺泡上皮细胞产生 IL-8 水平的影响.中华医学杂志,2006;86(15):1075-1076。
17 Alvarez RD, Canessa CM, Fyfe, et al. Structure and regulation of amiloride-sensitive sodium channel . Annu Rev Physiol, 2000, 62: 573-594.
18 Kellenberger S,Hoffmann Pochon N, Gautschi H, et al .On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
19 Carr S, Cantor JP, Cowan S,et al. Utilizing the peritoneum for gas exchange with oxygenated perfluorocarbons. American College of Surgeons 90th Annual Cliniacl Congress, New Orleans,2004.
20 Carr S, Cantor JP, Rao AS, et al. Peritoneal perfusion with Oxygenated Perfluorocarbon Augments Systemic Oxygenation. Chest,2006,130:402-411.
21 王晓光,赵晓巍,张健鹏. 全氟化碳雾化吸入对急性呼吸窘迫综合症猪气体交换、呼吸力学和血流动力学的影响.中华结核和呼吸杂志 2006;29(2):104-108.
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 M,Buhrow L,Wiedemann H.Perflubron decreases cytokine production by human alveolar macrophages.Crit Care Med,1997,25:2045–2047.
24 Woods CM,Gerald N,Elisabeth K,et al.Perflubron attenuates neutrophil adhesion to activated endothelial cells in vitro.Am J Physiol Lung Cell Mol Physiol,2000,278(5):L1008–L1017.
25 A Lazrak, Samanta K,Venetsanou, et al. Modification of biophysical properties of lung epithelial Na+ channels by dexamethasone. Am J Physiol 2000,279:762-770.
26 Thomassen MJ, Buhrow LT, Wiedemann HP. Perflubron decreases inflammatory cytokine production by human alveolar macrophages[J]. Crit Care Med,1997,25(12):2045-2047.
27 马利波,刘文. 全氟化碳的临床应用及其并发症. 眼科,2003,12(3):182-186.
28 刘书盈,樊毫军,张健鹏. 全身应用全氟化碳防护内毒素致急性肺损伤可行性研究. 山东医药,2006;46(34):18-19.
29 樊毫军,刘书盈,张健鹏.全氟化碳腹腔注射对内毒素肺损伤大鼠中性粒细胞肺侵润的抑制作用.中华结核与呼吸杂志 2006;29(10):674-678.
1 樊毫军,刘书盈,张健鹏,等.静脉注射内毒素致大鼠急性肺损伤模型的病理生理学指标评价。中国危重病急救医学,2006,18(8):485-487;
2 Matthay MA , Clerici C , Saumon G, et al. Active fluid clearance from the distal air spaces of the lung . J Appl Physiol 2002, 93: 1533-1541.
3 Berthiaume Y, Folkesson HG, Matthay MA,et al. Lung edema clearance: 20 years of progress: invited review: alveolar edema fluid clearance in the injued lung. Am J Physiol, 2002, 93(6):2207-2213.
4 Norlin A, Lu LN,Guggino SE ,et al. Contribution of amiloride insensitive pathways to alveolar fluid clearance in adult rats. J Appl Physiol, 2001,90(4):1489-1496.
5 张秋金,李银平,李檀实.肺泡上皮细胞功能特性与内毒素性急性肺损伤.中国危重病急救医学,2005,17(6):382-384.
6 Alvarez RD, Canessa CM, Fyfe GK, et al ,Structure and regulation of amiloride-sensitive sodium channel . Annu Rev Physiol, 2000, 62: 573-594.
7 Kellenberger S,Hoffmann Pochon N, Gautschi H, et al ,On the molecular basis of ion peimeation in the epithelial Na+ channel. J Gen Physiol ,1999,114:13-30.
8 Richard, Ramminger , Inglis, et al. O2 can raise fetal pneumocyte Na + conductance without affecting-ENaC-mRNAabundance.Biochem-Biophys-Res-Commun. 2003, 305(3): 671-676;
9 Matthay MA, Folkesson HG, Clerici C. Lung epithelial fluid transport and the resolution of pulmonary edema. Phsiol Rev 2002, 82(3):569-600;
10 Mairbaurl H, Mayer K, Kim KJ, et al. Hypoxia decreases active Na+ transport across primary rat alveolar epithelial cell monolayers. Am J Physiol Lung Cell Mol Physiol, 2002, 282(4):L659-665;
11 Chen XJ,Eaton DC,Jain L.Beta-adrenergic regulation of amiloride-sensitive lung sodium channels.Am J Physiol Lung Cell Mol Physiol,2002,282:L609-L620.
12 Dobbs LG, Gonzalez R , Matthay MA, et al. Highly water permeable type Ⅰ alveolar epithelial cells confer high water permeability between the airspace and vasculature in rat lung. Proc Nalt Acad Sci USA ,1998,95:2991-2996.
13 Matalon S, Lazrak A, Lucky J,et al. Lung clearance 20years lf progress inveted review :Biophysical properties of sodem channels in lung alveolar epithelial cells.J Appl Physiol , 2002, 93:1852-1859;
14 Mustafa SB, Geronimo RJ, et al. postatal glucocorticoids induce alpha-ENaCformation and regulate glucocorticoids receptors in the pretern rabbit lung. Am Physiol Lung Cell Mol physiol,2004, 286(1):L73-80.
15 宋志芳,谢伟,单慧敏,等.糖皮质激素治疗大肠杆菌致急性呼吸窘迫综合症的实验研究,中国危重病急救医学,2006,18(12):716-720.
16 Suzuki S, Matsuda Y, Sugawara T, et al.Effects of carbenoxolone on alveolar fluid clearance and lung inflammation in the rat. Crit Care Med, 2004, 32: 1910-1915.
17 Caldwell,Boucher, Stutts M.J. Neutrophil elastase activates near-silent epithelial Na+ channels and increases airway epithelial Na + transport. Am.J.Physiol Lung Cell Mol.Physiol.2005,288:813-819.
18 Matthay MA, Fukuda N, Frank J, et al. Alveolar epithelial barrier: role in lung fluid balance in clinical lung injury.Clinic Chest Med, 2000,21:477-490.
19 Benos DJ, Awayda MS,Ismailov, et al. Structure and function of amiloride-sensitive epithelial Na+ channels. J Membrane Biol, 1995,143:1-18.
20 McRitchie DI, Isowa N, Edelson JD, et al. Production of tumour necrosis factor by primary cultured rat alveolarepithelial cells. Cytokine, 2000,12: 644–654.
21 Suter PM, Suter S, Girardin E, et al. High bronchoalveolar levels of tumor necrosis factor and its inhibitors, interleukin-1, interferon, and elastase, in patients with adult respiratory distress syndrome after trauma, shock, or sepsis. Am Rev Respir Dis, 1992 145: 1016–1022.
22 Dagenais A,Frechette R,Yamagata Y,et al.Downregulation of EnaC activity and expression by TNF-alpha in alveolar epithelial cells.Am J Physiol Lung Cell Mol Physiol,2004,286:L301-L311.
23 Frank J,Roux J,Kawakatsu H,et al.Transforming growth factor –beta1 decreases expression of the epithelial sodium channel alphaENaC and alveolar epithelial vectorial sodium and fluid transport via ERK1/2-dependentmechnism.J Biol Chem,2003,278(45):43939-43950.
24 Towne JE,Harrod KS, Krane CM ,et al. Decreased expression of aquaporin (AQP)1 and AQP5 in mouse lung after acute viral infection. Am J Respir Cell Mol Biol, 2000, 22: 34-44.
25 Dagenais A, Gosselin D, Guilbault C, et al. Modulation of epithelial sodium channel expression in mouse lung infected with Pseudomonas aeruginosa. Respir. Res, 2005,6(6): 2.