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目的观察淋巴细胞缺陷对内毒素血症小鼠腹腔巨噬细胞活化的影响。方法采用腹腔注射脂多糖(LPS)建立Balb/c小鼠和T、B细胞缺陷的重症联合免疫缺陷(SCID)小鼠内毒素血症模型,ELISA检测2种小鼠腹腔灌洗液TNF-α和IL-10水平,实时荧光定量PCR检测腹腔巨噬细胞(peritoneal macrophage,PMa)TNF-α、IL-10及丝裂原蛋白激酶磷酸酶-1(mito-gen-activated protein kinase phosphatase-1,MKP-1)mRNA表达;ELISA检测Balb/c及SCID小鼠PMa体外刺激后细胞因子分泌情况。结果 LPS注射后1 h,SCID小鼠腹腔灌洗液TNF-α水平高于Balb/c小鼠,注射后3 h,IL-10水平低于Balb/c小鼠;LPS注射前及注射后,SCID小鼠PMa TNF-αmRNA表达高于Balb/c小鼠PMa,IL-10 mRNA表达低于Balb/c小鼠PMa;体外实验LPS刺激下,SCID小鼠PMa较Balb/c小鼠PMa分泌更多的TNF-α,IL-10的分泌却偏少。LPS注射前及注射后,Balb/c小鼠PMa MKP-1 mRNA表达均明显高于SCID小鼠。结论淋巴细胞缺陷导致内毒素血症小鼠腹腔巨噬细胞活性的增加,淋巴细胞抑制巨噬细胞的活化并可能调控其发育及成熟;MKP-1表达的减少可能是淋巴细胞缺陷导致腹腔巨噬细胞活性增加的分子机制之一。
Objective To observe the effects of lymphocyte defects on the activation of peritoneal macrophages in endotoxemic mice. Methods The model of severe combined immunodeficiency (SCID) mouse endotoxemia was established by intraperitoneal injection of lipopolysaccharide (LPS) in Balb / c mice and T and B cells. The levels of TNF-α And IL-10 levels were detected by real-time fluorescence quantitative PCR. The expressions of TNF-α, IL-10 and mito-gen-activated protein kinase phosphatase-1 in peritoneal macrophage (PMa) MKP-1) mRNA expression was detected by enzyme-linked immunosorbent assay (ELISA). The secretion of cytokines in Balb / c and SCID mice stimulated with PMa in vitro was detected. Results At 1 h after LPS injection, the level of TNF-α in peritoneal lavage fluid of SCID mice was higher than that of Balb / c mice at 3 h after injection of LPS, and the level of IL-10 was lower than that of Balb / c mice 3 h after LPS injection. Before and after LPS injection, The expression of PMa TNF-α mRNA in SCID mice was higher than that in Balb / c mice, and the expression of IL-10 mRNA was lower in Balb / c mice than in Balb / c mice More TNF-α, IL-10 secretion is less than normal. Before and after LPS injection, the expression of PMa MKP-1 mRNA in Balb / c mice was significantly higher than that in SCID mice. Conclusions Lymphocyte defects lead to the increase of peritoneal macrophage activity in endotoxemia mice. Lymphocytes inhibit the activation of macrophages and may regulate their development and maturation. The decrease of MKP-1 expression may be caused by lymphocyte defects leading to peritoneal macrophages One of the molecular mechanisms of increased cell viability.