人巨细胞病毒感染新生儿血清和脑脊液细胞因子的变化

Changes of cytokines in serum and cerebrospinal fluid of newborns infected with human cytomegalovirus

目的应用蛋白芯片检测人巨细胞病毒(HCMV)感染新生儿血清和脑脊液中的细胞因子,探讨HCMV感染导致血清和脑脊液中特异性细胞因子的变化,为HCMV感染引起神经系统损伤的程度预判提供依据。方法收集2016年6月至2017年12月中国医科大学附属盛京医院4例HCMV感染且具有中枢神经系统(CNS)损伤新生儿(HCMV感染组)的血清和脑脊液,4例无CNS感染性疾病新生儿(对照组)的血清和脑脊液,应用蛋白芯片筛选HCMV感染组与对照组相比血清及脑脊液中差异表达因子。进一步扩大样本收集30例HCMV感染组和30例对照组新生儿脑脊液,采用酶联免疫吸附法(ELISA)验证差异因子的表达。结果蛋白芯片分析发现,HCMV感染组新生儿脑脊液与对照组相比,在29种细胞因子的筛选中发现有3种差异表达因子,分别为脂肪细胞补体相关蛋白(Acrp30)、白细胞介素-1α(IL-1α)及基质金属蛋白酶-3(MMP3),均P<0.05。HCMV感染组新生儿血清与对照组比较,未见差异细胞因子表达。ELISA验证结果显示,HCMV感染组新生儿脑脊液中,Acrp30表达量显著高于对照组[(39.76±2.01) ng/L比(7.75±0.10) ng/L,t=87.09,P<0.001],MMP3的表达量高于对照组[(1.40±2.13) ng/L比(0.18±0.45) ng/L,t=3.07,P=0.003],而IL-1α显著低于对照组[(2.36±0.99) ng/L比(2.91±0.78) ng/L,t=2.39,P=0.020],差异均有统计学意义。结论HCMV感染新生儿脑脊液细胞因子的变化可能为HCMV的CNS损伤程度预后判断提供可靠参考指标,可协助临床对新生儿及时给予相应治疗,从而改善预后。

Objective To investigate the molecules of cytokine in the serum and cerebrospinal fluid of newborns infected with human cytomegalovirus (HCMV) by using protein chip technology and to analyze the changes of specific cytokine in serum and cerebrospinal fluid caused by HCMV infection, in order to provide a reliable index for predicting nervous system injury caused by HCMV infection. MethodsSerum and cerebrospinal fluid in 4 newborns with HCMV infection and central nervous system injury (HCMV-infected group), and 4 newborns without HCMV infection and central nervous system infection (control group) were collected in Shengjing Hospital of China Medical University from June 2016 to December 2017, and protein chip was used to screen the differentially expressed cytokines in newborns serum and cerebrospinal fluid.The samples were further expanded to collect cerebrospinal fluid from 30 newborns HCMV infection group and 30 newborns in the control group, and the expression of differentially proteins was verified by adopting enzyme linked immunosorbent assay(ELISA) method. ResultsThe results of protein chip analysis showed that newborns in HCMV infection group, compared with the control group, had 3 differentially expressed cytokines in the cerebrospinal fluid sample: adipocyte complement-related protein of 30 kD(Acrp30), interleukin-1 alpha(IL-1α), and matrix metallo protein-3(MMP3)(all P<0.05). Newborns in the HCMV-infected group, compared with the control group, had no differential cytokine expression in the serum.The results of ELISA showed that expression of Acrp30 was significantly higher in the cerebrospinal fluid of newborns with HCMV infection and central nervous system injury [(39.76±2.01) ng/L vs.(7.75±0.10) ng/L, t=87.09, P<0.001], and MMP3 expression was higher than that of control group [(1.40±2.13) ng/L vs.(0.18±0.45) ng/L, t=3.07, P=0.003], while the expression of IL-1α was significantly lower than that of the control group [(2.36±0.99) ng/L vs.(2.91±0.78) ng/L, t=2.39, P=0.020], and the differences were statistically significant. ConclusionsThe changes of cytokine in cerebrospinal fluid of HCMV infected newborn children may provide a reliable index for predicting injury degree of central nervous system in HCMV, and may further assist clinicians to give timely and appropriate treatment to newborns, and further assist clinicians to improve the prognosis for newborns.