Different expressions of Nogo-B1 and Nogo-B2 in mouse heart microvascular endothelial cell dysfunction induced by lysophosphatidylcholine

Abstract To investigate Nogo-B expression changes in mouse heart microvascular endothelial H5V cell line induced by lysophosphatidylcholine. Cells were incubated with different concentrations of lysoPC for the same incubation time and with 10 μmol/l lysoPC at different incubation times. Protein and mRNA expression levels of Nogo-B1 and Nogo-B2 were measured with Western blotting and semiquantitative RT-PCR, respectively. Nogo-B1 protein was detected in normal H5V cells by Western blotting. When H5V cells were incubated with lysoPC, Nogo-B1 protein level decreased, and the lowest point fell to 20% of the original level induced by 20 μmol/l lysoPC for 24 h. Incubation of H5V cells with lysoPC of different concentrations or at different time points caused little change in Nogo-B1 mRNA expression level, except for a 50% decrease in 20 μmol/l lysoPC at 24 h, while a transient change was observed in Nogo-B2 mRNA level. These results demonstrate that Nogo-B1 protein expression could be down-regulated with increasing concentrations of lysoPC and lapse of incubation time, though no mRNA transcription down-regulation occurred. However, mRNA expression level of Nogo-B2 showed a transient up-regulation induced by lysoPC. We conclude that the two subtypes of Nogo-B may play different roles in the endothelial cell injury process.