连续性血液净化对多器官功能障碍幼猪T辅助细胞免疫功能影响的机制研究

目的 研究连续性血液净化(CBP)对多器官功能障碍(MODS)幼猪在疾病不同时期外周血Th1、Th2细胞数量及其细胞培养上清液中细胞因子(IFN-γ、TNF-α、IL-12,IL-10、IL-4)水平的影响.方法 24只幼猪随机分为对照组和CBP干预组,每组12只.内毒素诱导多器官功能障碍.CBP主要采用连续性静脉静脉血液透析滤过(continuous venovenous hemodiafiltration,CVVHDF).造模完成后为0 h,MODS对照组在造模后0、2、4、6 h进行检测;血液净化干预组造模后即开始CVVHDF,分别在0 h及CVVHDF干预2 h、4 h、6 h进行检测.在各时点采用免疫磁珠分离两组幼猪外周血T辅助细胞,采用流式细胞仪检测Th1、Th2细胞数目,ELISA检测各时点T细胞培养上清液中IFN-γ、TNF-α、IL-12、IL-10、IL-4的表达水平.结果 幼猪诱导MODS模型后,辅助性T细胞分泌炎症因子TNF-α明显上升,而促进Th1细胞分化的因子IL-12和Th1细胞因子IFN-γ和调节因子IL-10呈下降趋势,Th2细胞因子IL-4则没有明显的变化,Th1/Th2比值下降;经过CVVHDF治疗,TNF-α和IL-4减少,IL-12、IFN-γ和IL-10在CVVHDF6 h后明显上升,Th1/Th2比值上升.结论 MODS幼猪模型经过CVVHDF治疗,有助于炎症因子的清除,增加机体的细胞免疫功能,恢复Th1/Th2的平衡.

The immune function influence of the CD4 + T helper cell after the treatment of continuous blood purification in a porcine model with multiple organ dysfunction syndrome

Objective To investigate the levels of Th1 and Th2 cell in the peripheral and the cytokines (IFN-γ,TNF-α,IL-12,IL-10,IL-4) in the culture medium at different time in the porcine model with multiple organ dysfunction syndrome(MODS) after continuous blood purification(CBP). Methods Twenty-four young porcines were given intravenous infusion of endotoxin to induce MODS, then were randomly divided into 2 groups:CBP group(n = 12) and MODS group without CBP(n = 12). Continuous venovenous hemodiafiltration (CVVHDF) was the main mode for CBP. At baseline, onset of MODS, and 2 h,4 h,6 h after treatment of CVVHDF,we picked the blood respectively to separate the T helper cell using the beads,detected the Th1 and Th2 cell by FACS and detected the cytokines (IFN-γ,TNF-α,IL-12,IL-10,IL-4) in the culture medium by ELISA. Results The MODS group showed a obvious rise in TNF-α,the drop tendency in IL-12,IFN-γ,IL-10,Th1/Th2 ratio, and no significant change in IL-4. After the treatment, the CBP group showed the drop in TNF-α and IL-4;and increase in IL-12,IFN-γ,IL-10 and Th1/Th2 ratio. Conclusion CBP helps to dear immune function,and restore the balance of Th1/Th2 in porcine MODS model.