南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (03): 428-.

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TSP-2对小鼠溃疡性结肠炎NF-κB DNA结合活性和表达的影响

赖明广,王立生,姚君,魏铖   

  • 出版日期:2013-03-20 发布日期:2013-03-20

TSP-2 suppresses the expression and DNA-binding activity of nuclear factor-κB p65 protein in mice with ulcerative colitis

  • Online:2013-03-20 Published:2013-03-20

摘要: 目的研究抗小鼠Toll样受体2(Toll-like receptor 2)胞外段(mTLR2ECD)单表位抗体TSP-2对溃疡性结肠炎小鼠肠粘膜
核因子NF-κB DNA结合活性和表达的影响。方法将实验小鼠随机分为4组:正常对照组、模型组、TSP-2治疗组和兔IgG组,其
中模型组、TSP-2治疗组和兔IgG组均给予5%葡聚糖硫酸钠7 d以制备溃疡性结肠炎小鼠模型,停用造模药后TSP-2治疗组和
兔IgG组给予后续干预治疗7 d。计算溃疡性结肠炎疾病活动指数(DAI),14 d后处死小鼠,取结肠行HE染色;利用ELISA法测
小鼠肠粘膜NF-κB P65 DNA结合活性;用Western blot 法测定NF-κB p65蛋白水平,并进行统计学分析。结果模型组NF-κB
p65 DNA结合活性明显高于对照组(P<0.05);模型组NF-κB P65 蛋白表达明显高于NC组(P<0.05);TSP-2治疗组小鼠第10~
14 天DAI明显低于模型组(P<0.05);TSP-2 治疗组NF-κB p65 DNA结合活性明显低于MD组(P<0.05);TSP-2 治疗组NF-κB
p65蛋白表达水平显著低于模型组(P<0.05);兔IgG组与模型组比较,实验结果无统计学意义(P>0.05)。结论TSP-2可以明显
抑制NF-κB表达和活性,通过调节肠道过度的免疫反应和抗炎作用,在溃疡性结肠炎治疗中发挥作用。

Abstract: Objective To observe the effect of TSP-2, the antibody of Toll-like receptor 2 extracellular domain, on the expression
and DNA-binding activity of nuclear factor-κB (NF-κB) p65 protein in mice with ulcerative colitis (UC). Methods Sixty BALB/c
mice were randomized equally into normal control group, UC model group, TSP-2 treatment group, and rabbit IgG treatment
group. In the latter 3 groups, the mice were fed with 5% DSS (C6H7Na3O14S3) solution for 7 days to induced UC, followed then
by treatment with daily injections of TSP-2 or rabbit IgG as appropriate for 7 days. The disease activity index was recorded
during the treatment. The colitis tissues were collected after the treatments for HE staining and detecting the expression and
DNA-binding activity of NF-κB p65 in the colon mucosa by Western blotting and ELISA. Results The DNA binding activity
and expressions of NF-κB P65 protein increased significantly in UC model group (P<0.05). TSP-2 treatment group significantly
decreased the disease activity index (P<0.05) and lowered the DNA-binding activity and expression of NF-κB P65 protein (P<
0.05) in the UC mouse models, while rabbit IgG produced no such effects (P>0.05). Conclusion TSP-2 can suppress the
DNA-binding activity and protein expressions of NF-κB P65 and regulate excessive immune response in the intestines to
ameliorate ulcerative colitis in mice.