[Studies on the expression of Src-suppressed C kinase substrate mRNA in pulmonary microvascular endothelial cells induced by lipopolysaccharide].
OBJECTIVE
To study the expression of Src-suppressed C kinase substrate (SSeCKS) mRNA in pulmonary microvascular endothelial cells(PMVEC) induced by lipopolysaccharide, and the interfering effect of methylprednisolone sodium succinate.


METHODS
Rat PMVEC (RPMVEC) were isolated and cultured in vitro, then grouped randomly according to different culture time and different dosage of LPS, and the expression of SSeCKS mRNA in RPMVEC of different groups were determined by reverse transcription polymerase chain reaction (RT-PCR).


RESULTS
Normally, the expression of the SSeCKS mRNA in RPMVEC was maintained on a low level, and it could be elevated by stimulation of LPS, and the changes in the expression exhibited in dosage and time dependent manner. After being stimulated with LPS for 1 hour, the extent of increase in SSeCKS mRNA in RPMVEC coincided with the increased dosage of LPS (control: 0.263+/-0.033; LPS of 0.1 mg/L: 0.529+/-0.066; LPS of 1 mg/L: 1.391+/-0.048; LPS of 10 mg/L: 2.339+/-0.055; LPS of 100 mg/L: 2.861+/-0.069, F=639.096, P<0.05). When RPMVEC were stimulated with 10 mg/L of LPS, the expression of SSeCKS mRNA began to increase at 0.5 hour, peaked at 1 hour, then decreased gradually, and it remained high even at 12 hours (control: 0.301+/-0.022; LPS 0.5 hour: 1.617+/-0.018; 1 hour: 2.378+/-0.031; 3 hours: 2.148+/-0.056; 6 hours: 1.322+/-0.042; 12 hours: 0.772+/-0.044, F=726.346, P<0.05); After methylprednisolone sodium succinate had been given, the increase of SSeCKS mRNA induced by LPS is inhibited (2.664+/-0.104 vs. 1.759+/-0.151, F=156.000, P<0.05).


CONCLUSIONS
(1) LPS could induce up-regulation of SSeCKS mRNA, the elevation is in time and dosage dependent manner. This indicate SSeCKS is related with LPS induced injury of RPMVEC. (2) Methylprednisolone sodium succinate inhibit the increase of SSeCKS mRNA induced by LPS.
Li
Hui
H
Department of Respiratary Medicine. The First Affiliated Hospital of Anhui Medical University, Hefei 230022, Anhui, China.
Sun
Geng-Yun
GY
Fei
Li-Ming
LM
Shen
Ji-Long
JL
D054758:A Kinase Anchor Proteins / Q000378:metabolism*; D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008168:Lung / Q000098:blood supply; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
D054758:A Kinase Anchor Proteins; C102361:Akap12 protein, rat; D018797:Cell Cycle Proteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger