人巨细胞病毒感染与冠心病患者Treg/Th17平衡及miR-155表达的关系

Relationship between human cytomegalovirus infection and regulatory T cells/helper T cells 17 balance and microRNA-155 expression in patients with coronary heart disease

目的分析人巨细胞病毒(HCMV)感染与冠心病患者外周血调节性T细胞(Treg)/辅助性T细胞17(Th17)平衡及微小RNA-155(miR-155)表达的相关性,探讨HCMV感染在冠心病发生发展中的作用。方法选择2018年1月至2019年1月期间保定市第四中心医院心内科住院部收治的冠心病患者160例,其中急性冠状动脉综合征(ACS)患者84例作为ACS组,稳定型心绞痛(SAP)76例作为SAP组,另选取同期到本院体检的非冠心病者80例作为对照组。采用酶联免疫吸附法(ELISA)检测血清HCMV-IgG、HCMV-IgM及血清白细胞介素-10(IL-10)、白细胞介素-17(IL-17)、转化生长因子-β(TGF-β)水平;实时荧光定量PCR法(RT-qPCR)检测血清miR-155表达;流式细胞术检测外周血Th17、Treg亚群比例,并计算Treg/Th17比值;Pearson分析ACS患者血清HCMV-IgM与miR-155、Treg/Th17相关性。结果与对照组比较,SAP组与ACS组HCMV-IgM阳性率及HCMV-IgM水平、HCMV-IgG阳性率及HCMV-IgG水平、Th17比例、IL-17水平升高,血清miR-155、IL-10、TGF-β水平、Treg比例、Treg/Th17比值降低,三组差异均有统计学意义(χ^(2)/F=35.744、38.400、36.526、151.238、114.878、439.705、185.429、384.825、127.762、53.129、533.803,均P<0.05)。与SAP组比较,ACS组HCMV-IgM阳性率及HCMV-IgM水平、Th17比例、IL-17水平升高,血清miR-155、IL-10、TGF-β水平、Treg比例、Treg/Th17比值降低,差异均有统计学意义(均P<0.05)。ACS组HCMV-IgM阳性患者血清miR-155水平、Treg比例、Treg/Th17比值低于HCMV-IgM阴性患者,Th17比例高于HCMV-IgM阴性患者,差异均有统计学意义(t=-9.430、-3.127、-5.005、2.366,均P<0.05);相关性分析显示,ACS患者血清HCMV-IgM与miR-155、Treg/Th17呈负相关(r=-0.647、-0.569,P<0.05)。结论HCMV感染可能参与冠心病的发生发展,与冠心病患者血清中miR-155、Treg/Th17比值有关。

Objective To analyze the correlation between human cytomegalovirus(HCMV)infection and the balance of regulatory T cell(Treg)/helper T cell 17(Th17),microRNA-155(miR-155)expression in peripheral blood of patients with coronary heart disease(CHD),and to explore the role of HCMV infection in the occurrence and development of CHD.Methods A total of 160 inpatients with CHD admitted to the cardiology department of the Fourth Central Hospital of Baoding City from January 2018 to January 2019 were selected,including 84 cases of acute coronary syndrome(ACS)as the ACS group and 76 cases of stable angina pectoris(SAP)as the SAP group.In addition,80 non-CHD patients underwent physical examination during the same period in the hospital were selected as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of HCMV-IgG,HCMVIgM,interleukin-10(IL-10),interleukin-17(IL-17)and transforming growth factor-β(TGF-β).Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect serum miR-155 expression.Flow cytometry was used to detect the proportions of Th17 cells and Treg cell subsets in peripheral blood,and calculate the ratio of Treg/Th17.Pearson analysis was used to evaluate the correlation between serum HCMV-IgM and miR-155,Treg/Th17 in ACS patients.Results Compared with the control group,the positive rate and level of HCMV-IgM,the positive rate and level of HCMV-IgG,Th17 proportion,and IL-17 level in the SAP group and the ACS group increased,the levels of serum miR-155,IL-10,TGF-β,Treg proportion,and Treg/Th17 ratio reduced,the differences were statistically significant among the three groups(χ^(2)/F=35.744,38.400,36.526,151.238,114.878,439.705,185.429,384.825,127.762,53.129,533.803;all P<0.05).Compared with the SAP group,the positive rate and level of HCMV-IgM,Th17 proportion,and IL-17 level in the ACS group increased,the levels of serum miR-155,IL-10,TGF-β,Treg proportion and Treg/Th17 ratio reduced,the differences were statistically significant(all P<0.05).In the ACS group,the serum miR-155 level,Treg proportion,and Treg/Th17 ratio of HCMV-IgM-positive patients were lower than those of HCMV-IgM-negative patients,and the Th17 proportion was higher than that of HCMV-IgM-negative patients,the differences were statistically significant(t=-9.430,-3.127,-5.005,2.366;all P<0.05).Correlation analysis showed that serum HCMV-IgM in ACS patients was negatively correlated with miR-155 and Treg/Th17 ratio(r=-0.647,-0.569;P<0.05).Conclusion HCMV infection may be involved in the occurrence and development of CHD,which is related to the serum miR-155 and Treg/Th17 ratio in patients with CHD.