中文摘要：

目的 探讨如何建立肾缺血再灌注损伤小鼠血清致肺微血管内皮细胞（PMVECs）损伤模型.方法 培养小鼠PMVECs,测定单层PMVECs标准电阻.待PMVECs生长成单层致密连接状态且达到标准电阻时,采用随机数字表法,将其分为4组（n=3）：正常小鼠血清组（NS组）和不同浓度肾缺血再灌注损伤小鼠血清组（IRS5组、IRS10组和IRS20组）.换成无血清内皮细胞培养基饥饿处理1h后,NS组上室和下室分别加入含20％正常小鼠血清的内皮细胞培养基0.8和0.2 ml进行培养;IRS5组、IRS10组和IRS20组上室和下室分别加入含5％、10％、20％肾缺血再灌注损伤小鼠血清的内皮细胞培养基0.8和0.2 ml进行培养.4组均在上室中加入100 μg/ml FITC-BSA 100μl作为通透物质.分别于培养3、6、9、12、15、18、21和24 h时,采用荧光白蛋白滤过法检测PMVECs的通透性,以通透系数（Pa）表示.结果 与NS组比较,IRS5组培养12和15 h时Pa升高,IRS10组和IRS20组培养6～24 h时Pa升高（P＜0.05）;与IRS5组比较,IRS10组培养21和24 h时Pa升高,IRS20组培养9～24 h时Pa升高（P＜0.05）;与IRS10组比较,IRS20组培养15～24 h时Pa升高（P＜0.05）.结论 10％和20％肾缺血再灌注损伤小鼠血清均可成功建立PMVECs损伤模型,其中20％血清时损伤更为明显.

英文摘要：

Objective To establish the model of serum-caused damage to pulmonary microvascular endothelial cells （PMVECs） of mice with renal ischemia-reperfusion （I/R） injury.Methods Mice PMVECs were cultured to measure the standard trans-endothelial electrical resistance （TER） in the monolayer of PMVECs.When PMVECs were cultured and arranged in compact monolayer and TER was achieved,they were divided into 4 groups （n =3 each） using a random number table：serum of normal mice group （NS group） and different concentrations （5%,10% and 20%） of serum of mice with renal I/R injury groups （IRS5 group,IRS10group and IRS20 group）.The PMVECs were cultured for 1 h in the serum-free endothelial culture medium.The 0.8 and 0.2 ml culture medium containing 20% serum of normal mice were then added to the upper and lower chambers,respectively,in group NS.The 0.8 and 0.2 ml culture medium containing 5%,10% and 20% serum of mice with renal I/R injury were then added to the upper and lower chambers in IRS5,IRS10 and IRS20 groups,respectively.100 μg/ml FITC-BSA 100 μl was added to the upper chamber in the four groups.At 3,6,9,12,15,18,21 and 24 h of incubation,the PMVEC monolayer permeability （apparent permeability coefficient,Pa） was detected.Results Compared with NS group,the Pa was significantly increased at 12 and 15 h of incubation in IRS5 group,and the Pa was increased at 6-24 h of incubation in IRS10 and IRS20 groups.Compared with IRS5 group,the Pa at 21 and 24 h in IRS10 group and at 9-24 h in IRS20 group were significantly increased.Conclusion Both 10% and 20% serum of mice with renal I/R injury can successfully establish the model of damage to PMVECs,and 20% serum causes a more severe damage.