AIM：To investigate the effect of tissue factor targeting peptide（TF-TP）on retinal pigment epithelium（RPE）cells tight junctions.METHODS：Cell counting kit-8（CCK-8）was used to measure the proliferation of ARPE-19...AIM：To investigate the effect of tissue factor targeting peptide（TF-TP）on retinal pigment epithelium（RPE）cells tight junctions.METHODS：Cell counting kit-8（CCK-8）was used to measure the proliferation of ARPE-19 cells.Expression of tight junction,ZO-1 in ARPE-19 cells was measured by Western blot and immunofluorescent staining.Western blot was also used to detect the expression of tissue factor（TF）.CEC Transmigration Assay was used to measure the migration of ARPE-19 cells.The transport of fluorescent markers [fluorescein isothiocyanate dextrans of 4,10,20（FD4,FD10,FD20） ]and the transepithelial electrical resistance（TEER）were used to measure in ARPE-19 cell RESULTS：CCK-8 assay showed that 5μmol/L TF-TP can inhibit ARPE-19 cells abnormally proliferation stimulated by lipopolysaccharide（LPS;P〈0.05）.LPS increased the transport of fluorescent markers（FD4,FD10,FD20）and decreased TEER levels in ARPE-19 cells,respectively,which were prevented by 5μmol/L TF-TP pretreatment（P〈0.05）. Furthermore,LPS significantly up-regulated the expression of TF and downregulated the expression of ZO-1（P〈0.05）in ARPE-19 cell which was inhibited by the TF-TP（P〈0.05）.In addition,TF-TP inhibited the abnormal migration induced by LPS in ARPE-19 cell（P〈0.05）.CONCLUSION：Our findings suggest that TF-TP suppressed proliferation and migration of ARPE-19 cells induced by LPS,and maintained the RPE tight junctions through inhibition of TF expression and increased expression of ZO-1.展开更多
基金Supported by Science and Technology Project of Guangzhou City(No.2014J4100035)the Project of the Third Affiliated Hospital of Guangzhou Medical University(No.2013Y06)
文摘AIM：To investigate the effect of tissue factor targeting peptide（TF-TP）on retinal pigment epithelium（RPE）cells tight junctions.METHODS：Cell counting kit-8（CCK-8）was used to measure the proliferation of ARPE-19 cells.Expression of tight junction,ZO-1 in ARPE-19 cells was measured by Western blot and immunofluorescent staining.Western blot was also used to detect the expression of tissue factor（TF）.CEC Transmigration Assay was used to measure the migration of ARPE-19 cells.The transport of fluorescent markers [fluorescein isothiocyanate dextrans of 4,10,20（FD4,FD10,FD20） ]and the transepithelial electrical resistance（TEER）were used to measure in ARPE-19 cell RESULTS：CCK-8 assay showed that 5μmol/L TF-TP can inhibit ARPE-19 cells abnormally proliferation stimulated by lipopolysaccharide（LPS;P〈0.05）.LPS increased the transport of fluorescent markers（FD4,FD10,FD20）and decreased TEER levels in ARPE-19 cells,respectively,which were prevented by 5μmol/L TF-TP pretreatment（P〈0.05）. Furthermore,LPS significantly up-regulated the expression of TF and downregulated the expression of ZO-1（P〈0.05）in ARPE-19 cell which was inhibited by the TF-TP（P〈0.05）.In addition,TF-TP inhibited the abnormal migration induced by LPS in ARPE-19 cell（P〈0.05）.CONCLUSION：Our findings suggest that TF-TP suppressed proliferation and migration of ARPE-19 cells induced by LPS,and maintained the RPE tight junctions through inhibition of TF expression and increased expression of ZO-1.
