尿毒症血液透析患者Treg/Th17功能失衡影响血管钙化因子BMP-2表达的研究

Effect of Treg/Th17 cells dysfunction on expression of vascular calcification factor BMP-2 in uremia patients with dialysis

目的探讨尿毒症血液透析患者Treg/Th17细胞功能是否发生失衡及其对血管钙化因子BMP-2表达的影响。方法选取66例尿毒症血液透析患者(MHD组)、30例尿毒症未并发心血管疾病且未透析患者(WHD组)和20例健康志愿者,将MHD组患者分为并发心血管疾病(MHD1,n=36)和未并发心血管疾病(MHD2,n=30)。流式细胞术测单个核细胞中Treg和Th17细胞比例,实时荧光定量PCR测Foxp3和ROR-γt mRNA表达,ELISA测单个核细胞上清液中炎症因子IL-17、IL-10和血管钙化因子BMP-2水平。结果与健康者相比,尿毒症患者出现Treg/Th17细胞功能失衡和BMP-2分泌异常,主要表现为Treg和Th17细胞比例、Foxp3和ROR-γt表达、IL-17和BMP-2水平均增高(P<0.05),而IL-10水平减低(P<0.05),这一失衡亦表现在WHD与MHD组、MHD1与MHD2组间。透析者IL-17、BMP-2水平均与Treg细胞比例呈负相关(P<0.05),而与Th17细胞比例呈正相关(P<0.05),其中BMP-2与IL-17、IL-10水平分别呈正相关和负相关(P<0.05)。结论尿毒症血液透析患者Treg/Th17细胞失衡使微炎症因子分泌异常,进而加剧血管钙化,导致加速性动脉粥样硬化和心血管疾病发生。

Objective To investigate Treg/Th17 cell dysfunction and their effects on expression of vascular calcification factor BMP-2 in uremia patients with dialysis.Methods Sixty-six uremia patients with dialysis（MHD group）,30 uremia patients neither treated by dialysis nor complicated with cardiovascular events（WHD group） and 20 healthy volunteers were enrolled.Patients of the MHD group were divided into two subgroups according to patients with（MHD1 group,n=36） or without（MHD2 group,n=30） cardiovascular events.Proportion of Treg and Th17 cells in mononuclear cells were measured by flow cytometry.Expression of ROR-γt and Foxp3 mRNA were measured by real-time PCR.Levels of IL-17,IL-10 and BMP-2 in supernatants of mononuclear cells were detected by ELISA.Results Uremia patients exhibited a dysfunction of Treg/Th17 and BMP-2 as compared with healthy controls,including a higher Treg/Th17 ratio,higher expression of Foxp3 and ROR-γt and higher concentrations of IL-17 and BMP-2.In contrast,IL-10 was significantly lower in the uremia patients（P〈0.05） than in the healthy controls.These dysfunctions also exhibited when comparison was performed between the WHD group and MHD group,and between the MHD1 group and MHD2 group,respectively.Concentrations of IL-17 and BMP-2 were negatively correlated with Treg cell proportion,but positively correlated with Th17 cell proportion;BMP-2 concentration was positively correlated with IL-17 concentration,but negatively correlated with IL-10 concentration.Conclusion Dysfunction of Treg/Th17 cells in uremia patients with dialysis can induce abnormal secretion of microinflammatory cytokines and aggravation of vascular calcification,leading to accelerated atherosclerosis and cardiovascular disease.