水杨酸柳氮磺胺吡啶对慢性结肠炎小鼠结肠组织中A20蛋白表达及脾脏中Treg比例的影响

Influence of sulfasalazine on colonic A20 protein expression and splenic Treg percent in mice with chronic colitis

目的:探讨水杨酸柳氮磺胺吡啶对慢性结肠炎小鼠结肠组织中A20蛋白表达及脾脏中Treg比例的影响。方法:18只C57BL/6小鼠分为对照组、模型组及水杨酸柳氮磺胺吡啶组,每组6只。对照组只给予正常饮水,模型组分别在第1、4、7周给予含20 g/L的葡聚糖硫酸钠饮水,水杨酸柳氮磺胺吡啶组则在模型组基础上在第3、4、18、39、46天给予水杨酸柳氮磺胺吡啶0.9 mg灌肠治疗。治疗后第1~7周观察小鼠一般状况变化,进行DAI评分;第7周末处死小鼠,取小鼠结肠组织分别采用HE染色和Western blot法进行病理学评分及A20蛋白表达测定,经抗CD4、CD25、Foxp3荧光抗体染色,流式细胞仪检测脾脏中Treg比例。结果:与对照组相比,模型组结肠组织病理学评分、DAI评分均增加;与模型组比较,水杨酸柳氮磺胺吡啶组结肠组织病理学评分、DAI评分均降低,结肠组织中A20蛋白相对表达量、脾脏中Treg比例增加(P<0.05)。结论:水杨酸柳氮磺胺吡啶可增加慢性结肠炎小鼠结肠组织中A20蛋白表达及脾脏中Treg比例。

Aim:To investigate influence of sulfasalazine on colonic A20 protein expression and splenic Treg percent in mice with chronic colitis.Methods:Eighteen C57BL/6 mice were randomly allocated into control group,model group and sulfasalazine group(n=6).Mice in control group received drinking water only.Mice in model group were administrated with 20 g/L dextran sulfate sodium in tap water on week 1,4,7 to induce chronic colitis model.Mice in sulfasalazine group received sulfasalazine(0.9 mg)by intrarectal instillation on day 3,4,18,39,46 during the colitis induction procedures.The general conditions of mice were observed daily for DAI score calculation.The colon tissue of the mice was taken after sacrifice on the 7th week,and the pathological observation and evaluation of A20 protein expression were performed by HE staining and Western blot.After staining with anti-CD4,CD25 and Foxp3 fluorescent antibodies,the Treg percent in the spleen was analyzed by flow cytometry.Results:Compared with the control group,the pathological score and DAI score in the model group were significantly higher(P<0.05).Compared with the model group,the pathological score and DAI score in the sulfasalazine group decreased significantly,and the expression level of A20 protein in colon tissue and the Treg percent in the spleen increased(P<0.05).Conclusion:Sulfasalazine could improve colonic A20 protein expression and splenic Treg percent in mice with chronic colitis.