Effect Of Microparticles In Patients With Ankylosing Spondylitis On Human Umbilical Vein Endothelial Cells In Vitro

Objective: To detect the levels of Microparticles(MPs)?Platelets microparticles(PMPs),Granulocyte microparticles(GMPs)?B-Lymphocyte microparticles(B-LMPs)?CD4+T-LMPs ?CD8+T-LMPs?and CD41a+CD62P+PMPs in plasma ankylosing spondylitis(AS)patients and healthy controls(HC).To evaluate the correlation of MPs ? PMPs ?CD41a+CD62P+PMPs ?GMPs?B-LMPs ?CD4+T-LMPs ?CD8+T-LMPs with clinical inflammatory parameters of AS patients.To determine whether MPs in plasma in AS patients can effect the cell viability of human umbilical vein endothelial cells(HUVEC).To investigate the impact of PMPs on gene expression of HUVEC in vitro,which including ICAM-1,VCAM-1,e NOS.Methods : Plasma were collected from 47 AS patients and 15 healthy volunteers.The levels of MPs,PMPs,GMPs,B-LMPs,CD4+T-LMPs,CD8+T-LMPs and CD41a+CD62P+PMPs in plasma in AS patients and HC were detected by flow cytometry.The clinical parameters including CRP,ESR,BASDAI were obtained.The effects of MPs on HUVEC cell viability were measured with Cell Counting Kit-8 assays.The gene expression of ICAM-1?VCAM-1and e NOS were determined with RT-q PCR in HUVEC after incubation with PMPs..Results:(1)The levels of MPs ?PMPs?CD41a+CD62P+PMPs ?GMPs in plasma from AS patients were higher than those in plasma from HC(p< 0.05).No significant differences were found between plasma from AS patients and HC in B-LMPs ?CD4+T-LMPs ?CD8+T-LMPs levels(p>0.05).(2)The levels of MPs ?PMPs?CD41a+CD62P+PMPs ?GMPs ?B-LMPs ?CD4+T-LMPs ?CD8+T-LMPs in plasma from AS patients were not correlated with ESR,CRP(p>0.05).(3)The levels of MPs in plasma of AS patients were positively correlated with BASDAI(r=0.5548,P=0.004),and the levels of PMPs in AS patients were positively correlated with BASDAI(r=0.5423,P=0.06)but the CD41a+CD62P+PMPs ? GMPs in AS patients were not correlated with BASDAI(p>0.05).(4)Compared with HC,the MPs in plasma from AS patients can significantly decreased cell viability after incubation with HUVEC for 24 hours(p<0.01).(5)when HUVECs were stimulated with PMPs for 24 hours,the levels of ICAM-1 m RNA are significantly incrased in AS patients when compared with blank and HC(p<0.01),but no significant difference between AS patients ? HC and Blank in the gene expression of e NOS ?VCAM-1 m RNA(p>0.05).Conclusion : The elevated levels of MPs and PMPs were correlated with disease activity of AS.The MPs in AS patients can decrease cell viability after incubation with HUVEC for 24 hours and PMPs can effect the gene expression ICAM-1m RNA of HUVEC which may increase cardiovascular risk in AS patients to some extent.