Influence Of Patients With Antiphospholipid Serum On The Expression Of Adhesion Molecule In Human Endothelial Cells And The Intervention Of Fluvastatin

Objective:To investigate the influence on ICAM-1,E-selectin,and VCAM-1 expression induced by the serum of patients with APS in human umbilical vein endothelial cells(HUVEC). As well as the intervention role and mechanism of fluvastatinMethods:HUVEC was primarily cultured by enzyme digestion procedure, the growth of the cells was observed under the phase-contrast microscope and all the cells were appraised though the eighth factor immunofluorescence.Choose the well-growth cells in the third and fouth generations for experiment. HUVEC were cultured with the following groups. (1) control group: with 1640 culture medium (2) APS group:with different concentration of serum from APS patients(10%,25%,50%,100%) for 24 hours. (3) Fluvastatin group:with different concentration of Fluvastatin (2.5,5 and 10μmol/L) combined with APS serum(50%) for 24 hours. (4) Mevalonate group:with APS serum(50%),fluvastatin (2.5,5 and 10μmol/L) and mevalonate (0.2mmol/L) mixed for 24 hours. mRNA level of ICAM-1,E-selectin,and VCAM-1 in the cells were measured by RT-PCR; the concentration of ICAM-1,E-selectin,and VCAM-1 in the supernatant were measured by ELISA and the amounts of NF-κΒp65 in each group was measured by western-blot.Results:1,Normal cells growed well with flat and polygon morphology,clear boundary,rich cytoplasm,round or oval nucleus. It showed a "paving stone" permutation with the increase of cell density. The immunofluorescence testimonied the positive expression of the eighth factor's relative antigen in endothelial cells.2,After stimulation of HUVEC with different concentration serum of APS, the concentration of ICAM-1,E-selectin and VCAM-1 in the supernatant, the content of ICAM-1,E-selectin,and VCAM-1 mRNA were signficiantly higher than the control group, and this effect was dose dependent(P<0.05).3,The concentration of ICAM-1,E-selectin and VCAM-1 in the supernatant, the content of ICAM-1,E-selectin and VCAM-1 mRNA and were dose-dependently decreased compared with control group after adding different concentration fluvastatin to HUVEC in the presence of serum of APS(50%). The amounts of NF-κΒp65 was consistent with the adhension molecules' levels(P<0.05).4,The concentration of ICAM-1,E-selectin and VCAM-1 in the supernatant, the content of ICAM-1,E-selectin and VCAM-1 mRNA and the amounts of NF-κΒp65 showed no significant difference from the 50% APS serum group Conclusions:1,The expression of ICAM-1,E-selectin and VCAM-1 in HUVEC cells can be induced by the serum of APS.2,Fluvastatin can inhibit the increased expression of ICAM-1,E-selectin and VCAM-1 in HUVEC induced by the serum of APS so as to prohibit the formation of thrombus and protect the endothelial cells within the antilipid effect.3,This protective effect maybe related to the NF-κΒpathway.