Effect of curcumin on HO-1 expression in endothelial progenitor cells from human umbilical vein blood

DING Hong; HUANG Wei-yi; SHEN Gui-dong

doi:10.3969/j.issn.1002-266X.2015.07.006

Abstract：Objective To observe the effects of curcumin ( CMN) on the HO-1 protein expression in endothelial pro-genitor cells ( EPCs ) from human umbilical vein blood and to investigate the mechanisms .Methods The mononuclear cells were isolated and cultured by density gradient centrifugation from human umbilical vein blood .The morphology of EPCs was observed with inverted microscope ,EPCs were also identified as adherent cells which were double positive for Dil -ac-LDL/FITC-UAE-1 by immumofluorescence method , and their expression of CD 133 was detected .EPCs were divided into four groups and incubated with 0, 5, 10 and 15μmol/L CMN respectively for 24 h.Then the HO-1 protein expression, the cell proliferation activity and the Lactate dehydrogenase ( LDH) activity in the supernatant were detected .EPCs were divid-ed into three groups and incubated with 0 μmol/L CMN (simple group), 15 μmol/L CMN (CMN group) and 15μmol/L CMN+1 μmol/L all-trans retinoic acid (ATRA) (CMN+ATRA group) respectively for 24 h.The HO-1 protein expres-sion was detected .Results Attached cells were observed after 3-4 days and cluster growth was noticed after 7 days.The immunofluorescence showed that the positive rate of CD 133 , the double staining positive rates of DiI-acLDL and FITC-UEA-1 were all more than 90%.With the increase of CMN concentration , the expression of HO-1 protein in the EPCs increased gradually (all P<0.05), and the cell proliferation increased significantly (all P<0.05), but the LDH activity in the su-pernatant did not change obviously (all P>0.05).Compared with simple medium group , the expression of HO-1 protein in the CMN and CMN+ATRA group increased (all P<0.05), and significant difference was found between the CMN and CMN+ATRA groups ( P<0.05).Conclusion CMN may effectively induce the high protein expression of HO-1 in EPCs, which is related with Nrf2/ARE signal pathway.