Establishment of model of serum-caused damage to pulmonary microvascular endothelial cells of mice with renal ischemia-reperfusion injury

Abstract

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. Key words: Reperfusion injury; Kidney; Serum; Endothelial cells