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[Studies on the expression of Src-suppressed C kinase substrate mRNA in pulmonary microvascular endothelial cells
Hui Li1, Geng-Yun Sun, Li-Ming Fei
1Department of Respiratary Medicine. The First Affiliated Hospital of Anhui Medical University, Hefei 230022, Anhui, China.
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).
Conclusion:
(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.
Insights
Lipopolysaccharide (LPS) up-regulates Src-suppressed C kinase substrate (SSeCKS) mRNA in pulmonary microvascular endothelial cells in a time- and dose-dependent manner. Methylprednisolone sodium succinate inhibits this LPS-induced increase.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Context:
- Pulmonary microvascular endothelial cells (PMVEC) play a critical role in lung inflammation.
- Lipopolysaccharide (LPS) is a potent inducer of inflammation and endothelial cell activation.
- Src-suppressed C kinase substrate (SSeCKS) is a protein kinase C substrate implicated in cellular signaling pathways.
Purpose:
- To investigate the expression pattern of SSeCKS mRNA in rat PMVEC (RPMVEC) following LPS stimulation.
- To determine the effect of methylprednisolone sodium succinate on LPS-induced SSeCKS mRNA expression.
Summary:
- LPS stimulation significantly increased SSeCKS mRNA expression in RPMVEC in a dose- and time-dependent manner.
- SSeCKS mRNA levels peaked at 1 hour post-LPS stimulation and remained elevated for up to 12 hours.
- Methylprednisolone sodium succinate treatment effectively inhibited the LPS-induced upregulation of SSeCKS mRNA.
Impact:
- These findings suggest that SSeCKS is involved in LPS-induced injury of RPMVEC.
- The inhibitory effect of methylprednisolone sodium succinate on SSeCKS mRNA expression may contribute to its protective role in LPS-induced lung injury.
