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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1μM sigma-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-α、IL-1β及抗炎因子IL-10水平的表达及fluvoxamine对细胞活性、炎症因子TNF-α、IL-1β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217.9μM(95%可区区间:188.5-251.8μM)。危重症患者的外周单核细胞分泌的促炎症因子TNF-α、IL-1β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-α、IL-1β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1μM BD1047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。
AIM: To investigate the effect of fluvoxamine, a sigma-1 receptor agonist, on the function of peripheral blood mononuclear cells in critically ill patients. Methods: Peripheral blood was collected from healthy subjects and critically ill patients. Peripheral blood mononuclear cells were isolated by centrifugation of lymphocytes. Isolated mononuclear cells were seeded in 96-well cell culture plates at a planting density of 106 per well. Then, different concentrations of fluvoxamine were added and co-cultured for 24 hours. In some plates, 1 μΜ sigma-1 receptor antagonist BD1047 was added simultaneously. The activity of TNF-α, IL-1β and IL-10 in peripheral blood mononuclear cells were detected by MTT, ELISA and fluorescence quantitative PCR, respectively. The effect of fluvoxamine on cell activity, inflammation TNF-α, IL-1β and anti-inflammatory cytokines IL-10. Western blot was used to detect the effect of fluvoxamine on sigma-1 receptor expression. RESULTS: Fluvoxamine affected peripheral mononuclear cell activity in critically ill patients at very high doses. The IC50 value was 217.9 μM (95% divisible interval: 188.5-251.8 μM). The levels of TNF-α and IL-1β secreted by peripheral blood mononuclear cells in critically ill patients were significantly higher than those in healthy subjects, while fluvoxamine inhibited the levels of TNF-α, IL-1β and other inflammatory cytokines in critically ill patients , Promote the expression of anti-inflammatory factor IL-10. But fluvoxamine has no effect on cytokines in healthy people. After pretreatment with 1 μM BD1047, fluvoxamine-treated monocytes were in the same activation state as those without fluvoxamine treatment. In addition, fluvoxamine and BD1047 did not affect sigma-1 receptor expression. Conclusion: Fluvoxamine can inhibit the activation of inflammation in critically ill patients, the effect may be related to its activation sigma-1 receptor.