ABSTRACT
Objective To study the changes of ICAM-1 expression in transplanted lung at the early stage after rat pulmonary transplantation.Methods The expression level of ICAM-1 protein and its mRNA in rat transplanted lung after 4 h of ventilation and reperfussion were detected by using immunohisto chemical method(SABC method)and semiquantitative reverse transcriptase polymerase reaction method in the experimental group(n=6).When donor lungs had been flushed with low potassium solution(LPDS) and preserved in 4.C LPDS for 12 h,left orthotopic pulmonary transplantation was performed.In control group (n=6), left pulmonary artery,pulmonary vein and bronchi were fully freed to be naked.Immunohistochemical test results were recorded in negative(-),suspicion(±),faint positive(+),positive(++)and intensive positive(+++).PCR products were separated by agrose gels and the density of the bands were determined by density scanning.Results The stained color of alveolar epithelial cells and pulmonary vascular endothdial cells in the transplanted lung was significantly more intensive in experimental group than in control group(P＜0.01) and the relative density values(ICAM-1/βactin)were also significandy higher in experimental group(0.837±0.044) than in control group (0.442±0.037),P＜0.01. Conclusions ICAM-1 expression in the transplanted lung was up-regulated in the early stage after pulmonary transplantation,which was related with the enhancement of ICAM-1 mRNA.
ABSTRACT
Intexcellular adhesion molecule-10CAM- 1) is an important moleade in aehating immune and inflammatory responses. It is found on the surface of hematopoietic and nonhematopoietic cells. It can act as an adhesive ligand for integrins such as LFA-1 (CD1&/CD11a) and MAC-1 (CD18/ CD11b). ICAM-1 is basally expressed in sigaificant amount on a limited number of cell types, including monocytes and endothelial ceRs. But it is inducible or upregulated by INF-r, IL-1p and TNF-a on many cell types. IL-4, a pleiotropic cytokine and mast cell differentiation factor, is upregulated in human allergic disease and stimulates expression of vascular adhesion molecule-1 (VCAM-1) in endothelial cells. IL-4 also promotes expression of surhce ICAM-1 in human mast cells and dermal fidroblasts. So in allergic rhinitis and asthma, IL-4 may be an important cytokine implicated in the pathogenesis of inflammation. We studied the effect of INF-r and IL-4 on expression of ICAM-1 in human nasal epithelial cells (HNEC). HNEC were prepared by primary culture method of monolayer culture of dissociated cells hom human inferior nasal turbinate mucosa. Nasal mucosa were obtained by partial turbinectomy of septal deviation patients. Primary cultured cells were charaterized as an epithelial cell type by indirect immuno-fluorescence assay using antilmdies against cytokeratin-pan, cytokeratin No. 8, vimentin and von Willebrand factor. Using fluorescence activated cell sorter (Coulter EI1TE), we analyzed the quantitative expression of ICAM-1 on HNEC. Treatment of HNEC with IFN-r (1ng/ml) for 24 hours caused about 8 fold increase of the surface ICAM-1 compared with constitutive expression by mean fluorescence intensity (MIF) but IL-4 had little effect. Theses foundings suggest that IFN-r is a potent ICAM-1 inducer in HNEC and further studies are necessary for the role of IL-4 on HNEC.