The Impact Of ET On Rat Peritoneal Mast Cell Empirical Study

Objective:To isolate and culture rat peritoneal mast cells in vitro,explore the effect of LPS on mast cell.Methods:Collecting the supernatant of rat PMCs,the morphologic features of PMCs were monitored by microscopy.PMCs vigor were observed with trypanblue dyeing,PMCs purity were observed with toluidine blue dyeing.PMS identited were divided into 5 groups:â‘ without LPS group;â‘¡0.5μg/ml LPS group;â‘¢1μg/ml LPS group;â‘£2μg/ml LPS group;⑤4μg/ml LPSgroup.The cells were cultured under the condition of 37℃,5%CO₂ for 2 hours,morphologic features and vigor of PMCs were monitored,PMCs degranulation were examed with toluidine blue dyeing.The histamine concentration in the supernatant of PMCs were detected by fluorometric method;The tryptase activities in the supernatant of PMCs were detected by BAPNA;The levels of IL-4,IL-10 in the supernatant of PMCs were detected by ELISA;The levels of TNF-αin the supernatant of PMCs were detected by ILIA.Results:1.The morphologic features of PMCs:most of rat PMC were demi-adherence growth by inverted microscopy,and round or oval-shape,intracytoplasm grains,compared to the no LPS treated group,PMCs in LPS treated groups showed a obvious degranulation,especialy in 1μg/ml LPS treated group.2.After 2 hours of PMCs together with different concentration LPS:1) The PMCs survival rates were above 90%in groups,which are not significantly dependent on the dose of LPS.2) The histamine and tryptase detected result display:PMCs in LPS treated groups increased significantly(P＜0.05),which were not significantly dependent on the doses of LPS(P＞0.05).3)Cytokine IL-4,IL-10 detected result display:PMCs in LPS treated groups increased significantly(P＜0.05),which were not significantly dependent on the doses of LPS(P＞0.05).4) Cytokine TNF-αdetected result display:PMCs in LPS treated groups increased significantly(P＜0.05),which were not significantly dependent on the doses of LPS(P＞0.05).Conclusion:compared to the no LPS treated group,PMCs in LPS treated groups have a obvious degranulation,which are not significantly dependent on the dose of LPS;The histamine,tryptase and IL-4,IL-10,TNF-αof PMCs release increased significantly,which are not significantly dependent on the dose of LPS.Indictioning that curing IETM add to inhibition degranulation,of PMC will acquired better effect.