溃结灵对溃疡性结肠炎大鼠结肠黏膜E1、UBC5、E3RSIκB mRNA与蛋白表达的影响  

Effect of Kuijieling on Colonic Mucosal E1, UBC5, E3RSIκB mRNA and Protein Expression in Ulcerative Colitis Rats

作　　者：邹娟[1] 李燕舞[1] 郭珍[1] 唐立海[1] 巫燕莉[1] 刘梓峰[1] 杜群[1] 

机构地区：[1]广州中医药大学脾胃研究所,广东广州510405

出　　处：《广州中医药大学学报》2014年第1期99-102,108,共5页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:30873421)

摘　　要：【目的】观察溃结灵对溃疡性结肠炎(UC)模型大鼠结肠黏膜泛素—蛋白酶体系统组分UBE1(E1)、UBC5(E2)、E3RSIκB(E3)基因和蛋白表达的影响。【方法】选用SD大鼠,随机分为6组:正常组,模型组,溃结灵高、中、低剂量组(剂量分别为18.3、9.2、4.6 g·kg-1·d-1),阳性对照组(柳氮磺胺吡啶,剂量为0.5 g·kg-1·d-1);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—聚合酶链反应(RT-PCR)法检测结肠黏膜E1、UBC5和E3RSIκB基因表达,酶联免疫吸附测定(ELISA)法检测结肠黏膜E1、UBC5和E3RSIκB蛋白含量。【结果】模型组大鼠结肠黏膜组织E1、UBC5 mRNA和蛋白相对表达量与正常组比较,差异均无统计学意义(P﹥0.05),溃结灵各剂量组E1、UBC5mRNA和蛋白相对表达量(除溃结灵高剂量组UBC5蛋白表达外)与模型组比较,差异均无统计学意义(P﹥0.05);模型组大鼠结肠黏膜组织E3RSIκB mRNA相对表达量显著高于正常组(P﹤0.05),溃结灵高、中剂量组E3RSIκB mRNA相对表达量显著低于模型组(P﹤0.05);模型组大鼠结肠黏膜组织E3RSIκB蛋白相对表达量显著高于正常组(P﹤0.05),溃结灵高、低剂量组E3RSIκB蛋白相对表达量显著低于模型组(P﹤0.05或P﹤0.01)。【结论】溃结灵抗UC作用的机理可能与抑制E3RSIκB的表达,阻止IκB的泛素化降解,进而抑制NF-κB的活化,减轻炎症反应有关。Objective To observe the effect of Kuijieling Decoction （KD） on the expression of ubiquitin- activating enzymes （El）, ubiquitin-conjugating enzyme UBC5 and ubiquitin ligase E3RSIKB mRNA and protein in colonic mucosa of ulcerative colitis （UC） model rats. Methods SD rats were randomized into 6 groups, namely normal group, model group, positive control group （treated with salazosulfapyridine 0.5 g·kg^-1·d^-1） , and high-, middle- and low-dose KD groups （in the dosage of 18.3, 9.2, 4.6 g·kg^-1·d^-1, respeetively） UC rat model was established with trinitro-benzene-sulfonic acid and were treated with the corresponding medicine according to the experimental design. El, UBC5 and E3RSIKB mRNA expression levels in colonie mueosa were tested by reverse transcription-polymerase ehain reaction （RT-PCR）, and El, UBC5 and E3RSIKB protein eontents were detected by enzyme-linked immunosorbent assay （ELISA） . Results The relative contents of E1 and UBC5 mRNA and protein in the colon of the model group had no statistically difference from those of the normal group （P〉0.05）, and the relative contents in the colon of high-, middle- and low-dose KD groups either did not differ from those of the model group （P〉0.05, except for UBC5 protein expression in high-dose KD group） .The relative contents of E3RSIKB mRNA and protein in the model group were significantly higher than those of the normal group （P〈0.05） . The relative eontent of E3RSIKB mRNA expression in high- and middle-dose KD group and the relative contents of E3RSIKB protein expression in high- and low-dose KD groups were significantly lower than those of the model group （P〈0,05 or P〈0.01） Conclusion The therapeutic mechanism of KD for UC degradation through inhibiting E3RSIKB expression, thus inflammation. may be related with the obstruction of ubiquitin to decrease the activation of NF-KB and relieve the

关 键 词：溃结灵 药理学 溃疡性结肠炎 中药疗法 基因表达调控 蛋白表达 疾病模型 动物 大鼠 

分 类 号：R285.5[医药卫生—中药学]