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[Role of RAGE in lipopolysaccharide-induced cytoskeletal changes in mouse pulmonary microvascular endothelial cells]
Xiao-Yan Zhou1, Wei-Jin Zhang, Qiao-Bing Huang
1Department of Pathophysiology of Southern Medical University, Key Laboratory of Shock and Microcirculation of Guangdong Province, Guangzhou 510515, China.E-mail: 602502708@qq.com.
Objective:
To investigate lipopolysaccharide (LPS)-induced changes of cytoskeletal filamentous actin in primary isolated pulmonary microvascular endothelial cells (PMVECs) from wild-type and RAGE knock-out mouse.
Methods:
The lungs of wild-type and RAGE knock-out mice were digested with collagenase type I to obtain endothelial cells purified by anti-CD31-coupled magnetic beads. The PMVEC identified by factor VIII labeling were stimulated with LPS at different concentrations and the changes of filamentous actin were observed by confocal microscopy.
Results:
The cultured primary cells showed typical endothelial cell phenotype as examined with factor VIII labeling. LPS stimulation caused rearrangement of the cytoskeletal filament F-actin in wild-type mouse PMVECs with stress fiber formation, but such changes were not obvious in RAGE knock-out mouse PMVECs.
Conclusion:
Mouse PMVECs of a high purity can be obtained by immune magnetic beads. RAGE is involved in LPS-induced destruction of mouse PMVEC cytoskeletons.
Insights
Lipopolysaccharide (LPS) causes cytoskeletal changes in mouse lung endothelial cells. The Receptor for Advanced Glycation End products (RAGE) is crucial in this LPS-induced F-actin rearrangement.
Area of Science:
- Cell Biology
- Immunology
- Pulmonary Medicine
Background:
- Pulmonary microvascular endothelial cells (PMVECs) play a critical role in lung function and inflammation.
- Lipopolysaccharide (LPS) is a potent inflammatory molecule that can affect endothelial cell integrity.
- The Receptor for Advanced Glycation End products (RAGE) is implicated in inflammatory responses.
Purpose of the Study:
- To investigate the impact of LPS on the F-actin cytoskeleton in primary mouse PMVECs.
- To determine the role of RAGE in LPS-induced cytoskeletal alterations in these cells.
Main Methods:
- Primary mouse PMVECs were isolated from wild-type and RAGE knock-out mice using collagenase digestion and magnetic bead purification.
- Cells were identified by factor VIII labeling.
- PMVECs were stimulated with varying concentrations of LPS, and F-actin cytoskeletal changes were visualized using confocal microscopy.
Main Results:
- Isolated PMVECs exhibited a typical endothelial cell phenotype.
- LPS stimulation induced significant F-actin rearrangement and stress fiber formation in wild-type PMVECs.
- These LPS-induced cytoskeletal changes were markedly attenuated in PMVECs from RAGE knock-out mice.
Conclusions:
- High-purity mouse PMVECs can be effectively isolated using immune magnetic beads.
- RAGE plays a significant role in the LPS-induced disruption of the mouse PMVEC cytoskeleton.
