Effecting Of Activated Platelets On Expression Of Cytokine IL-1β And IL-6 In Cultured Endothelial Cells

ObjectiveTo analyze the influence of platelets activated by ADP on expressing ofinflammatory cytokines (IL-1βand IL-6) in human umbilical vein endothelial cells(HUVECs) cultured in vitro. To explore the role of activated platelets playing ininflammatory diseases.Methods1 Peripheral blood from 30 healthy people were taken and platelet suspensionwere prepared with the method used by Ahnadi CE etc. 0.8μmol/l (finalconcentration) ADP were used to activate the platelets on conditions ofroom-temperature 20min. Mean platelet component(MPC) were detected by Advia120 Hematology system before and after intervention of ADP, and level of plateletactivation were evaluated.2 Frozen HUVECs (ECV304) was revived and subcultured. Mixed plateletsuspension was prepared with peripheral blood from healthy people and divided intothe following 4 experimental groups: (1) activated platelets group: achieved bymixing 0.8μmol/l (final concentration) ADP and platelets for 20min at roomtemperature and incubating with HUVECs. (2) Resting platelets group: achieved byresting platelets suspension for 20min at room temperature and incubating withHUVECs. (3) ADP control group. (4) Blank control group. Cells from each groupwere incubated for 12 hours at 37℃in circumstance of 5%CO₂. RNA was extractedwith TRNzol. Expressions of IL-1βand IL-6mRNA in endothelial cell were detectedby RT-PCR technology.Results1 Paired-Samples T test was performed to test the difference of two levels ofMPC achieved before and after activating platelets for 20 min by ADP with finalconcentration of 0.8μmol/l. t= 32.24, P＜0.01. The difference was statistically significant. Therefore, the research showed that platelets could be activated under theexperimental condition.2 When cultured with HUVECs, activated platelets could stimulate expressingof inflammatory cytokines IL-1βand IL-6mRNA on endothelial cell. While bothresting platelets and ADP with the same,content could not stimulate expressing ofIL-1βand IL-6mRNA.Conclusion:Activated platelets could involve in inflammatory reaction through inducinginflammatory cytokines IL-1βand IL-6 expressing on endothelial cell. This kind ofplatlet-endothlium reaction may be an important pathomechanism in someinflammatory diseases.