阻塞性睡眠呼吸暂停低通气综合征患者外周血CD4^+T细胞PD-1、CTLA-4表达及VEGF的变化研究

The expressions of CTLA-4 and PD-1 on CD4^+ T cells and the level of plasma VEGF in patients with obstructive sleep apnea hypopnea syndrome

目的探讨与肿瘤发生发展相关的CD4^+ T细胞、细胞毒性T淋巴细胞相关抗原-4(cytotoxic T-lymphocyte antigen 4,CTLA-4)、程序性死亡蛋白-1(programmed cell death-1,PD-1)及血管内皮生长因子(vascular endothelial growth factor,VEGF)在阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者中的变化。方法纳入2017年1月至2018年1月广东省人民医院睡眠呼吸障碍中心的首诊患者47例,根据多导睡眠监测(PSG)检查结果分为对照组、轻中度OSAHS组及重度OSAHS组,其中对照组17例,男10例,女7例,平均年龄(54±12)岁;轻中度OSAHS组15例,男9例,女6例,平均年龄(54±12)岁;重度OSAHS组15例,男10例,女5例,平均年龄(56±13)岁。采集静脉血,分离血浆及细胞,使用流式细胞术检测外周血CD4^+ T细胞表面PD-1和CTLA-4的表达,用酶联免疫吸附实验测定血浆VEGF的浓度。结果对照组、轻中度OSAHS组及重度OSAHS组CD4^+ T细胞比例分别为(38±8)%、(35±8)%和(38±6)%(F=1.228,P>0.05),CD4^+ T细胞CTLA-4的表达分别为1.13%(0.59%~1.78%)、0.45%(0.16%~1.43%)和0.87%(0.47%~1.46%),差异无统计学意义(H=2.205,P>0.05),CD4^+ T细胞PD-1的表达分别为4.24%(2.12%~6.03%)、3.54%(2.69%~5.09%)和3.31%(1.67%~8.25%),差异无统计学意义(H=0.541,P>0.05)。对照组、轻中度OSAHS组及重度OSAHS组外周血VEGF浓度分别为(395±88)、(453±108)及(546±199)ng/L,差异有统计学意义(F=4.827,P=0.013),重度OSAHS组明显高于对照组(P<0.01)。VEGF浓度与夜间最低SpO2呈负相关(rs=-0.480,P=0.001),与AHI(rs=0.403,P=0.005)、ODI(rs=0.378,P=0.010)、SpO2≤90%(%TST)(rs=0.547,P=0.000 3)均呈正相关。结论OSAHS患者CD4^+ T细胞PD-1及CTLA-4的表达无显著变化,但有下降趋势;VEGF表达上调,且随着严重程度和夜间缺氧程度加重而增高。

Objective CD4^+T cells,cytotoxic T-lymphocyte antigen 4 (CTLA-4),programmed cell death-1 (PD-1) and vascular endothelial growth factor (VEGF) are associated with cancer development.The aim of the present study was to investigate the expression of CTLA-4,PD-1 and VEGF in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods From January 2017 to January 2018,a total of 47 first-visit outpatients were recruited in the Sleep and Respiratory Disorder Center of Guangdong Provincial People′s Hospital,and were divided into control group (N=17,mean age 54±12 years),mild-to-moderate OSAHS group (N=15,mean age 54±12 years) and severe OSAHS group (N=15,mean age 56±13 years).Venous blood was collected,plasma and cells were isolated,the expressions of PD-1 and CTLA-4 on the surface of CD4^+T cells were detected by flow cytometry,and plasma VEGF was measured by enzyme linked immunosorbent assay.Results The proportion of CD4^+T cells in control group,mild-to-moderate OSAHS group and severe OSAHS group were respectively(38±8)%,(35±8)% and (38±6)%(F=1.228,P>0.05).The expression of CTLA-4 on CD4^+T cells were respectively [1.13 (0.59~1.78)]%,[0.45 (0.16~1.43)]% and [0.87(0.47~1.46)]%(H=2.205,P>0.05).The expression of PD-1 on CD4^+T cells were respectively [4.24 (2.12~6.03)]%,[3.54(2.69~5.09)]% and [3.31(1.67~8.25)]%(H=0.541,P>0.05).The concentrations of VEGF in control group,mild-to-moderate OSAHS group and severe OSAHS group were statistically different [(395.16±87.78) ng/L vs (452.85±107.97) ng/L vs (546.42±199.27) ng/L,F=4.827,P=0.013].Compared with the control group,VEGF concentration was significantly increased in the severe OSAHS group(P<0.01).VEGF concentration was correlated negatively with the lowest SpO2 (r s=-0.480,P=0.001),but positively with apnea-hypopnea index(r s=0.403,P=0.005),oxygen desaturation index (r s=0.378,P=0.010) and proportion of SpO2 less than or equal to 90% of total sleep time(r s=0.547,P=0.000 3).ConclusionThere was no significant difference of PD-1 and CTLA-4 expression on CD4^+T cells in patients with and without OSAHS.The expression of VEGF was elevated in OSAHS patients,and increased with the severity of OSAHS and hypoxia.