寨卡病毒感染人神经胶质瘤细胞所诱导的细胞因子表达水平的变化

Changes of cytokine expression levels in human glioma cells infected with ZIKA virus

目的研究寨卡病毒(ZIKV)感染人神经胶质瘤细胞(U251)所导致的细胞因子分泌的变化,探讨ZIKV影响神经系统功能紊乱的机制。方法建立ZIKV感染U251细胞模型,收获感染后第1、3、5、7天细胞培养上清液,采用悬浮芯片系统(Bio-Plex~? 200)Luminex多重检测技术同时定量检测细胞因子IL-4、IL-5、IL-6、TNF-α和IL-12P70的浓度。结果细胞因子IL-4的浓度水平低于检测范围(<0.001 pg/mL);病毒感染组分泌IL-5、IL-6、TNF-α和IL-12P70(169.9±12.1、7 747.9±949.2、29.0±0.5和93.3±2.0 pg/mL)的浓度水平均高于对照组(分别为124.6±8.6、2 652.2±386.0、25.5±0.8和85.4±1.8 pg/mL),且随着病毒感染时间的增加而分泌增加;在所有检测的细胞因子中,最显著表达的细胞因子是IL-6。结论 ZIKV感染U251细胞可分泌浓度水平相对较高的促炎性细胞因子,促炎细胞因子可能是影响神经发育的关键因素。

Objective To study the changes of cytokine secretion induced by ZIKA virus(ZIKV) infection of human glioma cells U251, and to reveal the mechanism of ZIKV affecting nervous system dysfunction. Methods A ZIKV-infected U251 cell model was established. Cell culture supernatants were harvested at the 1 st、3 rd、5 th and 7 th d after infection, and the suspension chip system(Bio-Plex~? 200) Luminex multiplex detection technology was used for quantitatively detecting the concentrations of cytokines IL-4、IL-5、IL-6、TNF-α and IL-12 P70. Results The concentration of cytokine IL-4 was lower than the detection range(<0.001 pg/mL). The levels of IL-5、IL-6、TNF-αand IL-12 P70(169.9±12.1,7 747.9±949.2,29.0±0.5 and 93.3±2.0 pg/mL,respectively)secreted by the virus infection group were higher than those of the control group(124.6±8.6 pg/mL,2 652.2±386.0 pg/mL,25.5±0.8 pg/mL and 85.4±1.8 pg/mL),and the secretion increased with the increasing of virus infection time. Of all the detected cytokines, the most significant expressed cytokine was IL-6. Conclusion ZIKV infected U251 cells can secrete relatively high levels of proinflammatory cytokines. Proinflammatory cytokines may be a key affecting factor for neural development.