Influence of lipopolysaccharide on growth of in vitro human coronary artery smooth muscle cells: an investigation on establishing in-stent restenosis inflammatory cell model

LI Hong-Mei; WANG Xian

doi:10.3969/j.issn.1674-4055.2016.01.07

Abstract：Objective To investigate the influence of lipopolysaccharide (LPS) in different doses on growth of in vitro human coronary artery smooth muscle cells (HCASMC) at different times, analyze non-toxic concentration range of LPS to HCASMC, and provide test data for establishing HCASMC inflammation activation model in studies related to restenosis after interventional therapy.Methods HCASMC was cultured in vitro for 3-5 generations, and seeded onto 96-well plates. The influences of LPS in different doses (0 μg/mL, 0.01 μg/mL, 0.1 μg/mL, 0.5 μg/mL, 1 μg/mL, 5 μg/mL, 10 μg/mL and 100 μg/mL) on viability of HCASMC at different time points (24 h,46 h,72 h) were detected by using MTT assay. The value A was detected by using ELISA at point of 492 nm, and cell viability=(value A of test group- value A of zero hole)/value A of control group.Results When LPS dose was lower than 100 μg/mL and interventional time was shorter than 48 h, there was no cytotoxicity observed to HCASMC and there was promotion effect on proliferation. When interventional time was longer than 48 h, there was significant cytotoxicity observed and viability of HCASMC would decrease as LPS dose increased. After intervention for 72 h, LPS had promotion effect on proliferation in doses from 0 μg/mL to 0.01 μg/mL, and mild inhibitory effect in doses from 0.1 μg/mL to 0.5 μg/mL but cytotoxicity was not significant. The toxic reaction gradually appeared when LPS dose was from 1 μg/mL to 100 μg/mL, and viability of HCASMC decreased as LPS dose increased. When LPS doses were the same, viability of HCASMC gradually decreased as interventional time was prolonged, and the longer the interventional time was, the more significant the decrease of HCASMC was.Conclusion LPS had no significant cytotoxicity when its dose is 0.1 μg/mL, and when its dose is 0.01 μg/mL, the proliferation of HCASMC will be more significant after intervention for 24 h, which can be taken as the best condition for establishing HCASMC inflammation activation model.