外源性白蛋白输入对ARDS小鼠肺血管通透性的相关研究
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摘要
目的:本研究采用腹腔注射内毒素建立ARDS小鼠模型,用EB(伊文思蓝)标记方法,研究外源性白蛋白输入对ARDS小鼠肺血管通透性和CRP、PCT的影响。
方法:1.采用腹腔注射5mg/kg、10mg/kg、15mg/kg3种不同浓度内毒素,并于4、8、16、24h4个时间点断颈取血(每次6只),采用ELASA法测定血清CRP和PCT;同时留取肺测湿、干重。2.采用腹腔注射内毒素(LPS)15mg/kg,制备ARDS模型,尾静脉注射0.05g/L荧光标记的牛血清白蛋白0.1ml/10g体质量,于注射后2、4、8、16和24h断颈处死(每次6只),取适量肺组织匀浆,荧光分光光度计测上清液的荧光值,对照标准曲线计算出各样本荧光白蛋白的量。3.制备ARDS模型后,分为未治疗组和治疗组,治疗组分别尾静脉注射0.5g/L人血清白蛋白0.1mL/10g体质量、50.0g/L人血清白蛋白0.1mL/10g体质量、60.0g/L羟乙基淀粉0.1mL/10g体质量。各小组取6只,分别于处死前30min尾静脉注射5.0g/L伊文思蓝0.1ml/10g体质量,于注射后4、8、16、24h断颈处死(每次6只),用改良伊文思蓝渗出法测定肺组织伊文思蓝漏出量。各组剩余6只小鼠于各时间点断颈取血测血清CRP、PCT,并留取肺测湿、干重。
结果:1.各致伤浓度组小鼠肺W/D从4h上升,8h达到高峰,16h有所下降,24h稍有恢复(P＜0.05)。随感染程度的加深和时间的延长,PCT逐渐升高(P＜0.05);CRP有在16h达到高峰,也有在24h达到高峰。2.荧光标记的牛血清白蛋白漏出量在4-24h内持续升高。3.与未治疗组比较,50.0g/L人血清白蛋白和60.0g/L羟乙基淀粉组伊文思蓝漏出量无明显差别(P＞0.05);CRP和PCT有所下降(P＜0.05);死亡率无明显变化;肺组织病理表现减轻。
结论:50.0g/L人血清白蛋白能降低早期ARDS肺含水量,降低CRP和PCT,减轻炎症反应,不增加肺血管通透性,有助于早期ARDS的治疗。
Objective:In this study,Lipopolysaccharide(LPS) was injected in mice to establish ARDS models by intraperitoneal.EB(Evans blue) labeling was used to research the influence of importation of exogenous albumin on pulmonary vascular permeability and CRP、PCT of ARDS models.
Methods:1.5mg/kg、10mg/kg、15mg/kg of LPS were injected in different mice by intraperitoneal.Mice were killed by break neck at 4、8、16、24h after injected(6 mice per). Serum was kept.ELASA was used to determine CRP and PCT,and wet/dry weight of the lung was also determined.2.15mg/kg of LPS was injected in different mice by intraperitoneal to establish ARDS models.0.05g/L of fluorescent labeling of bovine serum albumin 0.1mL/10g body mass was given by tail vein injection and mice were killed by break neck at 2、4、8、16、24h after injected(6 mice per).Take appropriate lung tissue,homogenate,We can know fluorescence value of supernatant by fluorescence spectrophotometer.Then,We can calculate the amount of albumin fluorescence of samples by control of standard curve.3.ARDS models were divided to injury but non-treated and treated groups.Treatad groups were given 0.5g/L of human serum albumin 0.1ml/10g body mass,50.0g/L of human serum albumin 0.1ml/10g body mass,60.0g/L hydroxyethyl starch by tail vein injection respectively.Check the group 6,5.0g/L Evans Blue(EB) was respectively injected in mice by tail vein at 30min before the end of the execution.Mice were killed by break neck at 4、8、16、24h after injected(6 mice per). Leakage of Lung tissue Evans blue was messured by modified EB exudation determination.Then,We killed the other 6 mice of different group by break neck at 4、8、16、24h after injected(6 mice per) to keep serum to determine CRP and PCT and to determine wet/dry weight of the lung.
Results:1.The W/D of mice lung increased from 4h,peak at 8h,decline at 16h and recovered slightly at 24h(P<0.05) in various concentrations of injury groups.With the deepening of degree of infection and extension of time,PCT increased gradually(P< 0.05);CRP peaked at 16h or 24h.2.The leakage of fluorescent labeling of bovine serum albumin rose in 4-24h sustainly.3.Compared with the untreated group,there was no difference in leakage of lung tissue EB and mortality rate in 50.0g/L of albumin group and 60.0g/L of hydroxyethyl starch group(P>0.05);the CRP and PCT in the two groups decreased;lung tissue pathology in the two groups improved.
Conclusion:50.0g/L albumin can reduce lung wet/dry weight in early ARDS,reduce CRP and PCT,reduce inflammatory response,not increase pulmonary vascular permeability and contribute to the treatment of early ARDS.
方法:1.采用腹腔注射5mg/kg、10mg/kg、15mg/kg3种不同浓度内毒素,并于4、8、16、24h4个时间点断颈取血(每次6只),采用ELASA法测定血清CRP和PCT;同时留取肺测湿、干重。2.采用腹腔注射内毒素(LPS)15mg/kg,制备ARDS模型,尾静脉注射0.05g/L荧光标记的牛血清白蛋白0.1ml/10g体质量,于注射后2、4、8、16和24h断颈处死(每次6只),取适量肺组织匀浆,荧光分光光度计测上清液的荧光值,对照标准曲线计算出各样本荧光白蛋白的量。3.制备ARDS模型后,分为未治疗组和治疗组,治疗组分别尾静脉注射0.5g/L人血清白蛋白0.1mL/10g体质量、50.0g/L人血清白蛋白0.1mL/10g体质量、60.0g/L羟乙基淀粉0.1mL/10g体质量。各小组取6只,分别于处死前30min尾静脉注射5.0g/L伊文思蓝0.1ml/10g体质量,于注射后4、8、16、24h断颈处死(每次6只),用改良伊文思蓝渗出法测定肺组织伊文思蓝漏出量。各组剩余6只小鼠于各时间点断颈取血测血清CRP、PCT,并留取肺测湿、干重。
结果:1.各致伤浓度组小鼠肺W/D从4h上升,8h达到高峰,16h有所下降,24h稍有恢复(P＜0.05)。随感染程度的加深和时间的延长,PCT逐渐升高(P＜0.05);CRP有在16h达到高峰,也有在24h达到高峰。2.荧光标记的牛血清白蛋白漏出量在4-24h内持续升高。3.与未治疗组比较,50.0g/L人血清白蛋白和60.0g/L羟乙基淀粉组伊文思蓝漏出量无明显差别(P＞0.05);CRP和PCT有所下降(P＜0.05);死亡率无明显变化;肺组织病理表现减轻。
结论:50.0g/L人血清白蛋白能降低早期ARDS肺含水量,降低CRP和PCT,减轻炎症反应,不增加肺血管通透性,有助于早期ARDS的治疗。
Objective:In this study,Lipopolysaccharide(LPS) was injected in mice to establish ARDS models by intraperitoneal.EB(Evans blue) labeling was used to research the influence of importation of exogenous albumin on pulmonary vascular permeability and CRP、PCT of ARDS models.
Methods:1.5mg/kg、10mg/kg、15mg/kg of LPS were injected in different mice by intraperitoneal.Mice were killed by break neck at 4、8、16、24h after injected(6 mice per). Serum was kept.ELASA was used to determine CRP and PCT,and wet/dry weight of the lung was also determined.2.15mg/kg of LPS was injected in different mice by intraperitoneal to establish ARDS models.0.05g/L of fluorescent labeling of bovine serum albumin 0.1mL/10g body mass was given by tail vein injection and mice were killed by break neck at 2、4、8、16、24h after injected(6 mice per).Take appropriate lung tissue,homogenate,We can know fluorescence value of supernatant by fluorescence spectrophotometer.Then,We can calculate the amount of albumin fluorescence of samples by control of standard curve.3.ARDS models were divided to injury but non-treated and treated groups.Treatad groups were given 0.5g/L of human serum albumin 0.1ml/10g body mass,50.0g/L of human serum albumin 0.1ml/10g body mass,60.0g/L hydroxyethyl starch by tail vein injection respectively.Check the group 6,5.0g/L Evans Blue(EB) was respectively injected in mice by tail vein at 30min before the end of the execution.Mice were killed by break neck at 4、8、16、24h after injected(6 mice per). Leakage of Lung tissue Evans blue was messured by modified EB exudation determination.Then,We killed the other 6 mice of different group by break neck at 4、8、16、24h after injected(6 mice per) to keep serum to determine CRP and PCT and to determine wet/dry weight of the lung.
Results:1.The W/D of mice lung increased from 4h,peak at 8h,decline at 16h and recovered slightly at 24h(P<0.05) in various concentrations of injury groups.With the deepening of degree of infection and extension of time,PCT increased gradually(P< 0.05);CRP peaked at 16h or 24h.2.The leakage of fluorescent labeling of bovine serum albumin rose in 4-24h sustainly.3.Compared with the untreated group,there was no difference in leakage of lung tissue EB and mortality rate in 50.0g/L of albumin group and 60.0g/L of hydroxyethyl starch group(P>0.05);the CRP and PCT in the two groups decreased;lung tissue pathology in the two groups improved.
Conclusion:50.0g/L albumin can reduce lung wet/dry weight in early ARDS,reduce CRP and PCT,reduce inflammatory response,not increase pulmonary vascular permeability and contribute to the treatment of early ARDS.
引文
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