Endothelial Cells Conditioned Medium IL-6on The Proliferation And ICMA-1、IL-8of The Vascular Smooth Muscle Cells

Objective:To investigate the influence of activated endothelial cells on the proliferation and the secretion of the vascular smooth muscle cells, and the mechanism between them.Method::Cultured HUVECs was treated with different doses and durations of TNF-a, To collect the supernatant fluid Enzyme-linked immunosorbent assay(ELISA) were used to measure IL-6production in HUVECs. TUNEL was used to detect the apoptosis of endothelial cells, the supernatant fluid which was treated with lOng/ml TNF-a for24h, called fluid A; the supernatant fluid which was treated with Ong/ml TNF-a for24h, called fluid B, to collect them, to filter with micropore film, The vascular smooth muscle cells were inoculated on cultivation plates and divided to4groups as follows①Activated group:to add fluid A and DMEM (1:1);②Antagonism group:Add the same to activated group and anti IL-6monoclonal antibody;③Control group:to add fluid B and DMEM (1:1)④Blank group:to add DMEM only. All the supernatant fluid was collected at the Oh and24h after grouping, then ELISA was used to measure the level of ICAM-1、IL-8. The proliferation of the vascular smooth muscle cells were detected by MTT.Results:TNF-a can induce endothelial IL-6production in vascular endothelial cells in a dose and time-dependently manner; TNF-a induced apoptosis of endothelial cells in a time-dependant manner, the OD value of the4groups were significant difference. At the end of the experiment, ICAM-1and IL-8were increased than the baseline values in4groups, the maximum increase was occur in Activated group.Conclusion:The TNF-a activated endothelial cell promote the vascular smooth muscle cells proliferation and the secretion of ICAM-1and IL-8by elevated IL-6releasing.