溃结灵对溃疡性结肠炎大鼠结肠黏膜泛素表达的影响

Effect of Kuijieling Decoction on the Expression of Ubiquitin in Colonic Mucosa of Ulcerative Colitis Model Rats

目的:观察溃疡性结肠炎(ulcerative coliotis,UC)大鼠模型结肠黏膜泛素基因和蛋白表达的变化,并探讨溃结灵对其的作用,进一步揭示溃结灵防治UC的作用机理。方法:采用三硝基苯磺酸(TNBS)法复制UC大鼠模型并进行不同剂量药物干预。采用逆转录PCR方法检测结肠黏膜泛素基因表达,Elisa方法检测结肠黏膜泛素蛋白的表达。结果:模型组泛素基因相对表达量高于正常组(P<0.05);溃结灵中、低剂量组泛素基因相对表达量低于模型组(P<0.05)。模型组泛素蛋白相对表达量高于正常组(P<0.05);溃结灵中、低剂量组泛素蛋白相对表达量均低于模型组(P<0.01,P<0.05)。结论:溃结灵对TNBS法UC大鼠模型结肠黏膜泛素基因和蛋白的表达均有下调作用,其作用机理可能为通过抑制IκB-α的泛素化降解,从而抑制NF-κB的活化,减轻炎症反应。

Objective To observe the effect of Kuijieling decoction（KD） on the expression of gene and protein of ubiquitin in colonic mucosa of rats with ulcerative colitis（UC） and explore its possible mechanism. Methods Trinitro-benzene-sulfonic acid（TNBS） method was used to establish UC rat model. After treatment with different dosages of KD, reverse transcription polymerase chain reaction（RT-PCR） was used to detect the expression of ubiquitin gene in colonic mucosa, and the expression of ubiquitin protein was detected by enzyme-linked immunoabsorbent assay（ELISA）. Results The expression of ubiquitin gene in model group was higher than that in normal group（P 〈 0.05）; and the relative expression of ubiquitin gene in middle and low dosage KD groups were both significantly lower than that in model group（P 〈 0.05）. The relative expression of ubiquitin protein in model group was notably higher than that in normal group（P〈 0.05）; and the relative expression of ubiquitin protein in middle and low dosage KD groups were both obviously lower than that in model group（P〈0.01, P〈0.05） Conclusion The relative expressions of the gene and protein of ubiquitin in colonic mucosa of UC model rats inducedby TNBS couldbe down-regulated after the treatment with KD. The mechanism might be that KD could inhibit the activation of NF-κB and reduce inflammatory reaction through preventing the ubiquitination degradation of IkB-α.