Different expressions of Nogo-B1 and Nogo-B2 in mouse heart microvascular endothelial cell dysfunction induced by lysophosphatidylcholine.
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 micromol/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 micromol/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 micromol/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.
Pan
Jing-Wei
JW
Department of Cardiology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
Zheng
Xing
X
Yang
Peng-Yuan
PY
Qin
Yong-Wen
YW
Rui
Yao-Cheng
YC
Ma
Li-Ping
LP
Zhou
Fei
F
Kang
Hui
H
D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005786:Gene Expression Regulation; D008244:Lysophosphatidylcholines / Q000378:metabolism*; D051379:Mice; D009185:Myelin Proteins / Q000096:biosynthesis*; D009206:Myocardium / Q000378:metabolism*; D000070798:Nogo Proteins; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013997:Time Factors; D014158:Transcription, Genetic
D008244:Lysophosphatidylcholines; D009185:Myelin Proteins; D000070798:Nogo Proteins; D012333:RNA, Messenger; C000603201:Rtn4 protein, mouse