IL-15 mutant/Fcγ2a融合蛋白对溃疡性结肠炎模型小鼠的治疗作用及其机制研究-东南大学学报医学版

IL-15 mutant/Fcγ2a融合蛋白对溃疡性结肠炎模型小鼠的治疗作用及其机制研究

单位：1. 东南大学医学院, 江苏南京 210009;
2. 同济大学附属东方医院胃肠外科, 上海 200120;
3. 东南大学附属中大医院普通外科, 江苏南京 210009;
4. 同济大学附属东方医院消化内科, 上海 200120

分类号：R574.62;R-33

出版年·卷·期（页码）：2016·35·第四期（559-564）

摘要：

目的：探讨IL-15 mutant/Fcγ2a融合蛋白（IL-15 mutant/Fc）对溃疡性结肠炎（UC）小鼠模型的治疗作用及其可能的作用机制。方法：5%葡聚糖硫酸钠（DSS）连续饮用制成UC模型，随机将30只小鼠分为对照组、IL-15 mutant/Fc治疗组、IgG2a治疗组，观察小鼠一般体重情况。造模完成后取结肠行大体病理观察，并行伊红染色，查看其病理学损伤水平。ELISA分析小鼠眼球血的IL-10、IL-15、TNF-α炎症因子的含量，RT-PCR测定结肠组织STAT3和STAT5 mRNA的表达，蛋白质印迹法测定磷酸化STAT5（p-STAT5）以及磷酸化AKT（p-AKT）蛋白的表达。结果：除对照组小鼠外，其他两组均出现一般情况及体重改变。而IL-15 mutant/Fc组小鼠的症状以及组织病理学评分均轻于IgG2a组。IL-15 mutant/Fc可改善IL-10、IL-15、TNF-α的分泌情况。IL-15 mutant/Fc抑制STAT5 mRNA、STAT3 mRNA表达以及p-STAT5、p-AKT蛋白的表达。结论：IL-15 mutant/Fc在体内具有改善模型UC小鼠病情的作用，它可能通过特异性的拮抗IL-15受体来干预STAT、AKT信号通路的信号转导而发挥其治疗作用。

Objective:To investigate the therapeutic effect and mechanisms of IL-15 mutant/Fcγ2a fusion protein(IL-15 mutant/Fc)in mice with ulcerative colitis(UC). Methods:Thirty mice were randomly divided into control group, IL-15 mutant/Fc treatment group and IgG2a treatment group.UC model was induced in the latter two groups by giving 5% dextran sulphate sodium in drinking water for 7 consecutive days. In the experiment, the general condition and weight of the mice were observed, and the protective effects were evaluated by macroscopic and pathologic examinations. Hematoxylin-eosin(HE)staining of the distal colon mucosa of mice in each group was performed to study the histopathological injury and histological scores. The concentration of inflammatory cytokines(IL-10, IL-15, TNF-α) in ocular blood of mice was determined by ELISA assay. Expression of STAT3 and STAT5 mRNA and expression of p-STAT5 and p-AKT protein were detected by reverse transcriptase polymerase chain reaction(RT-PCR) and Western Blot respectively. Results:Compared with the control group, mice in other two groups showed poor general conditions and changes in body weight, but the general conditions and histopathological score of mice in IL-15 mutant/Fc treatment group were better than those in IgG2a treatment group. The secretion disorder of pro-inflammatory cytokines and anti-inflammatory cytokine could be partially alleviated by injection of IL-15mutant/Fc demonstrated by increasing IL-10 and decreasing IL-15 and TNF-α. The levels of STAT3 and STAT5 mRNA, and of p-STAT5 and p-AKT protein in IL-15 mutant/Fc treatment group were obviously lower than those in IgG2a treatment group(P<0.05). Conclusion:As a specific IL-15 receptor antagonist, IL-15 mutant/Fc can achieve its therapeutic effect on UC by intervening the signaling pathway through regulating the mRNA or protein expressions of key signal molecules, such as STAT and AKT.

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