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目的：研究sigma-1 受体的激动剂fluvoxamine 对危重症患者外周血单核细胞功能的影响。方法：收集健康体检者和危重症患者外周血，并采用淋巴细胞分离液离心，分离外周血单核细胞。将分离的单核细胞种植于96 孔细胞培养板中，种植密度为106/ 孔。然后，加入不同的浓度的fluvoxamine，共培养24 小时。在部分培养板中，同时加入1 uMsigma-1 受体拮抗剂BD1047。采用 MTT、ELISA 和荧光定量PCR 分别检测体检者与危重患者外周血单核细胞的细胞活性、炎症因子TNF-alpha、IL-1beta及抗炎因子 IL-10 水平的表达及fluvoxamine 对细胞活性、炎症因子TNF-alpha、IL-1beta及抗炎因子IL-10 水平的影响；采用Western blot 检测 fluvoxamine 对sigma-1 受体表达的影响。结果：Fluvoxamine 在极高剂量下才影响危重症患者外周单核细胞的活性。其IC50 值为 217.9 uM( 95%可区区间: 188.5-251.8 uM)。危重症患者的外周单核细胞分泌的促炎症因子TNF-alpha、IL-1beta的水平明显高于健康体检患者，而fluvoxamine 能够抑制危重症患者单核细胞中TNF-alpha、IL-1beta等炎症因子的表达，促进抗炎症因子IL-10 的表达。但是 fluvoxamine 对健康人群的细胞因子没有影响。经1 uMBD1047 预处理以后，fluvoxamine 处理的单核细胞与未经fluvoxamine 处理的细胞处于同样的活化状态。另外，fluvoxamine 和BD1047 并不影响sigma-1受体的表达。结论：Fluvoxamine 能够抑制危重症患者的炎症活化状态，其效应可能与其激活sigma-1 受体有关。

英文摘要:

Objective:To investigate the immuno-modulatory effect of fluvoxamine, a sigma-1 receptor agonist, on the peripheral monocytes of critical ill patients.Methods:The peripheral blood was collected from the healthy volunteers and the critical ill patients. The peripheral monocytes were isolated using lymphocyte separation mediums through centrifugation. The isolated peripheral monocytes was seeded in the 96-well culture plates at the density of 106 per well. Then, fluvoxamine was added and incubated for 24 hours at the different concentrations. In some cases, 1 uM BD1047, a sigma-1 receptor antagonist, was added. The levels of cytokines in the culture medium, such as TNF-alpha, IL-1beta and IL-10, were quantified with ELISA and the corresponding intra-cytosolic mRNA expression were examined by real time-PCR. The expression of sigma-1 receptor was examined with western blot.Results:Fluvoxamine at extreme high concentration inhibited the viability of peripheral monocytes. The IC50 was 217.9 uM(95%confidential interval: 188.5-251.8 滋M). The monocytes in the critical ill patients were over-activated to the stimulation of lipopolysaccharide, compared with the healthy volunteers. The mediumlevels of pro-inflammatory factors (TNF-alpha, IL-1beta) in the critical ill patients were higher than those of the healthy volunteers. Fluvoxamine at the concentration of 2.5-10 nM significantly inhibited the production of TNF-alpha and IL-1beta, and promoted the production of an anti-inflammatory factor, IL-10, in the critical ill patients. But, fluvoxamine didn't change the expressions of TNF-alpha, IL-1beta, and IL-10 in the healthy volunteers. In addition, BD1047, a sigma-1 receptor antagonist, ameliorated the immuno-modulatory effect of fluvoxamine. When being pretreated with 1 uM BD1047, the monocytes treated by fluvoxamine were kept at the similar over-activated levels with those without the treatment of fluvoxamine. In addition, fluvoxamine and BD1047 didn't alter the expression of sigma-1 receptor in the peripheral monocytes.Conclusion:Fluvoxamine could modulate the inflammatory response of the critical ill patients, and this activity depended on the activation of sigma-1 receptor.

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