ABSTRACT
Objective:To study the effects of LPS and TNF-? on the expression of SSeCKS and morphology as well as cytoskeleton of endothelial cell, so as to explore the role of SSeCKS in cell morphology changes.Methods:The cultured Bovine pulmonary artery endothelial cells(BPAEC) was induced by LPS, TNF-? and the expression of SSeCKS was detected by in situ hybridization,Western blot and immunohistology. Immunofluorescent staining method with confocal laser-scanning fluorescence microscope was used to observe the effects of LPS and TNF-? on the morphology of endothelial cells and the organisation of SSeCKS as well as cytoskeleton.Results:Firstly, we found that TNF-? could induce the expression of SSeCKS in a concentration and time dependent manner , meanwhile LPS had no effects on SSeCKS expression. Secondly, it was observed that LPS and TNF-? induced reorganization of F-actin and SSeCKS in endothelial cell. Thirdly,PKC inhibitor Ro-31-8220 reversed the effect of LPS,TNF-? on F-actin and SSeCKS in endothelial cells.Conclusion:The results demonstrate that TNF-? could induce endothelial cell to express SSeCKS; PKC plays a role in the reorganization of SSeCKS and F-actin in endothelial cells induced by LPS and TNF-?; the results suggest that the mechanism for reorganization of cytoskeleton induced by LPS, TNF-? be partially related to the SSeCKS of ECs.
ABSTRACT
Objective To study the localizations and changes of mRNA for nNOS-interacting DHHC domain-containing protein with dendritic mRNA(NIDD) in normal and injured peripheral nerves. Methods Real-time fluorescence quantitative PCR(FQ-PCR) and a combination of in situ hybridization and indirect immunofluorescence were used to detect the localizations and changes of mRNA for nNOS and NIDD. Results It was found that nNOS and NIDD were mainly localized in Schwann cells(SCs) of normal sciatic nerve,and the expression peaked in crushed sections of sciatic nerve in the 2nd week after injury and in proximate or distal transected stumps in the 1st week after injury respectively.Expressions of mRNA for nNOS and NIDD were detected in the perinuclear plasm of primary SCs as well.Conclusion The increase in mRNA for NIDD in SCs may play an important role in axonal regeneration after nerve injury.