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目的:观察肠道病毒71型(EV71)感染小鼠外周血IL-6,TNF-α水平的变化及脑细胞凋亡情况,以探讨EV71中枢损害可能的发病机制。方法:建立小鼠EV71感染模型,小鼠根据不同处理因素分为对照组和实验组,用RT-PCR检测感染1天、3天、5天、7天的小鼠脑组织的EV71RNA;分别收集外周血,用ELISA法检测血清细胞因子IL-6,TNF-α的含量;制备单细胞悬液,采用AnnexinV/PI双染结合流式细胞术(FCM)测定小鼠脑细胞的凋亡情况。应用方差分析进行统计学处理。结果(1)实验组于感染第5天和第7天在脑组织中可检测到病毒RNA,对照组脑组织未检测到病毒RNA。(2)实验组血清IL-6,TNF-α水平明显高于对照组(P<0.05),并于24-72h达高峰,此后逐渐下降,于第7天时基本降至正常。(3)实验组小鼠于感染第3天开始出现脑细胞凋亡,于第5天达高峰,第7天时脑细胞凋亡反而降低。结论:EV71具有嗜神经性,EV71感染能促进单核-巨噬细胞分泌IL-6,TNF-α介导全身炎症反应,EV71感染过程中存在脑细胞凋亡现象,可能是其中枢损害的机制之一。
Objective: To observe the changes of IL-6 and TNF-α in peripheral blood of mice infected with enterovirus 71 (EV71) and the apoptosis of brain cells to explore the possible pathogenesis of EV71 central lesion. Methods: The mice model of EV71 infection was established. The mice were divided into control group and experimental group according to different treatment factors. The EV71 RNA in brain tissues of mice infected with day 1, day 3, day 5 and day 7 were detected by RT-PCR. The levels of IL-6 and TNF-α in serum were measured by ELISA. Single cell suspension was prepared and the apoptosis of mouse brain cells was determined by Annexin V / PI double staining combined with flow cytometry (FCM). Analysis of variance was used for statistical analysis. Results (1) In the experimental group, viral RNA was detected in the brain tissue on the 5th and 7th days of infection, but no viral RNA was detected in the control group. (2) The levels of serum IL-6 and TNF-α in the experimental group were significantly higher than those in the control group (P <0.05), and peaked at 24-72h, then decreased gradually, and then decreased to normal on the 7th day. (3) In the experimental group, the apoptosis of brain cells began to appear on the 3rd day of infection and peaked on the 5th day. On the 7th day, the apoptosis of brain cells decreased. CONCLUSION: EV71 has neuropathicity. EV71 infection can promote the secretion of IL-6 by monocyte-macrophage. TNF-α mediates the systemic inflammatory response. The apoptosis of brain cells during EV71 infection may be the mechanism of central nervous system damage one.