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摘要:
目的 探讨内毒素对小鼠脂钙蛋白2 (lipocalin 2, LCN2)及其相关细胞因子表达水平的影响。 方法 C57BL/6小鼠随机分为4组(n=8), 实验组分别予10、20、30 mg/kg内毒素腹腔内注射, 对照组仅注射等容积生理盐水。观察24 h后留取盲肠和血清, 酶联免疫吸附法和蛋白免疫印迹法测定白细胞介素(interleukin, IL)-1β、IL-6、IL-17A、LCN2水平, 实时PCR相对定量肠组织中LCN2 mRNA。 结果 与对照组比较, 大剂量内毒素组(20 mg/kg和30 mg/kg)的血清IL-6、LCN2与肠组织IL-1β、IL-6、IL-17A、LCN2水平均显著增高(P < 0.05)。与10 mg/kg组比较, 30 mg/kg组的血清IL-6、LCN2以及肠组织IL-1β、IL-17A、LCN2水平均显著增高(P < 0.05)。对照组及10、20、30 mg/kg内毒素组肠组织LCN2 mRNA相对定量值分别为1.548±0.417、3.184±0.716、4.875±0.547、7.489±0.820, 两两比较差异均有统计学意义(P < 0.05)。 结论 内毒素可剂量依赖性地引起小鼠肠组织LCN2表达水平增高; 血清IL-6与LCN2水平以及肠组织IL-1β、IL-17A与LCN2水平之间存在一定的相关性。 Abstract:Objective To explore the influence of an endotoxin lipopolysaccharide (LPS) injection on the expressions of lipocalin 2 (LCN2) and its relative cytokines in mice. Methods C57BL/6 mice were randomly divided into 3 groups (n=8 each) according to the serial LPS dosages of 10, 20, and 30 mg/kg. The control group (n=8) received only isotonic saline injection. Twenty-four hours later, cecum and blood samples were collected, and enzyme-linked immunosorbent assay (ELISA) and Western blot were performed to detect the serum levels of interleukin(IL)-1β, IL-6, IL-17A, and LCN2, and real-time polymerase chain reaction (PCR) to determine the LCN2 mRNA level in cecum lysate. Results The serum levels of IL-6 and LCN2 and the levels of IL-1β, IL-6, IL-17A, and LCN2 in cecum lysate in two large-dosage LPS groups (20 mg/kg and 30 mg/kg) were significantly higher than their counterparts in control group (P < 0.05). Compared with the 10 mg/kg LPS group, the levels of IL-6 and LCN2 in serum and the levels of IL-1β, IL-17A, and LCN2 in cecum lysate significantly increased in the 30 mg/kg LPS group (P < 0.05). 2-ΔΔCT was used to indicate fold changes of LCN2 mRNA in intestine. The results expressed as mean±standard deviation were as follows:1.548±0.417 for con-trol group, 3.184±0.716 for 10 mg/kg LPS group, 4.875±0.547 for 20 mg/kg LPS group, and 7.489±0.820 for 30 mg/kg LPS group, and the difference was significant between either two groups (P < 0.05). Conclusions LPS dose-dependently increases the level of LCN2 expression in cecum. The serum IL-6 level as well as the cecum IL-1β and IL-17A levels have certain correlations with the LCN2 level. -
Key words:
- endotoxin /
- lipocalin 2 /
- cytokine
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图 2 盲肠组织LCN2表达水平(以β-肌动蛋白为内参照)
A.蛋白免疫印迹显影条带;B.显影区带光密度值相对定量,数据资料以中位数和取值范围表示;C.实时荧光PCR定量肠组织LCN2 mRNA水平,以各实验组相对于对照组的变化倍数表示,数据资料以均数和标准差表示Mr:相对分子质量;LCN2:同图 1;与对照组比较,*P<0.05;与10 mg/kg内毒素组比较,†P<0.05;与20 mg/kg内毒素组比较,‡P<0.05
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