RESUMO
AIM:To investigate the expression and function of store-operated calcium channels ( SOCC) in human circulating fibrocytes.METHODS:Peripheral blood mononuclear cells ( PBMCs) were isolated and cultured in ser-um-free media.After 7 d, the PBMCs differentiated into fibrocytes.RT-PCR and real-time PCR were performed to deter-mine the mRNA expression of ORAI1-3 and STIM1-2 in the fibrocytes.SOCC inhibitor SKF-96365 was used to elucidate the role of SOCC in the differentiation of fibrocytes.RESULTS:The results of real-time PCR showed that the mRNA ex-pression of ORAI1-3 and STIM1-2 was positive in cultured fibrocytes.SKF-96365 (10μmol/L) significantly inhibited the differentiation of fibrocytes.CONCLUSION:SOCC-related proteins ORAI1-3 and STIM1-2 are abundantly expressed in the fibrocytes, and may play an important role in the differentiation of these cells.
RESUMO
Aim To isolate and characterize the human circulating fibrocytes from human peripheral blood and explore the effects of curcumin on human circulating fi-brocytes.Methods The cells were isolated and puri-fied by density gradient centrifugation,and identified by flow cytometry and immunocytochemistry .Then , CCK-8 and flow cytometry were used to study the effect of curcumin on the proliferation as well as COL I ex-pression of human circulating fibrocytes,respectively. Results After being isolated the cells expressed CD34,CD45 and COLⅠ,among which 79.7% were both CD45 and collagen I positive,typical of human circulating fibrocytes.Curcumin could exert regulatory effects on proliferation of human circulating fibrocytes. Exposure of the cells to curcumin for as short as 24 hours promoted their growth,while prolonged treatment (72 h ) significantly inhibited cell propagation and downregulated the COLⅠ levels,best manifested at a concentration as high as 20 μmol · L-1 .Conclusion The proliferation of cells and COLⅠexpressions can be effectively inhibited by curcumin with the prolonged action period and high concentrations.