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Based on the safe range of lipopolysaccharide(LPS) density explored in the preliminary study, we investigated the effects of different ratios of LPS at different points on the expression of inflammatory cytokines in human coronary artery smooth muscle cells(HCASMC) cultured in vitro.
The 3-5 generations of HCASMC was respectively seeded onto 6-well plates, then co-incubated with different concentration of LPS (0,0.01,0.1,0.5,1,and 10μg/ml) at different points (0,6,24,and48h). Then the expression of inflammatory cytokines were determined via enzyme-linked immunosorbent assay(ELISA).
The effect of LPS on the expression of inflammatory cytokines in HCASMC were related with the density and action-time. The dose of (0.01,0.1,0.5,1,10)μg/ml LPS with action-time of (24,48h) could significantly induce the expression of IL-1β(P<0.05). 0.5μg/ml LPS with action-time of (6,24,48h) could induce the expression of IL-6 and TNF-α(P<0.05), while 1μg/ml LPS with action-time of (24,48h) could induce the expression of IL-6 and TNF-α significantly(P<0.05).
Under certain conditions, LPS can activate HCASMC pathway and increase secretion of specific inflammatory factors. The dose of (0.5,1)μg/ml LPS with action-time of (24,48h) can significantly induce the expression of IL-1β, IL-6 and TNF-α, which can be used in the experimental study of HCASMC cultured in the inflammation condition.