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【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 on the gene and protein expression of UBE1 (E1), UBC5 (E2) and E3RSIκB (E3) in the colonic mucosa of UC in ulcerative colitis model rats. 【Methods】 SD rats were randomly divided into 6 groups: normal group, model group, Kuijieling high, medium and low dose groups (doses were 18.3, 9.2 and 4.6 g · kg-1 · d-1, The positive control group (sulfasalazine, 0.5 g · kg-1 · d-1) was used. The UC rat model was transplanted with trinitrobenzene sulfonic acid (TNBS) and the drug intervention was performed at different doses. The expressions of E1, UBC5 and E3RSIκB in colonic mucosa were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the contents of E1, UBC5 and E3RSIκB in colonic mucosa were detected by enzyme linked immunosorbent assay (ELISA). 【Results】 The mRNA and protein expressions of E1 and UBC5 in colonic mucosa in model group were not significantly different from those in normal group (P> 0.05). The relative expressions of E1 and UBC5 mRNA and protein (P <0.05). The relative expression of E3RSIκB mRNA in colonic mucosa in model group was significantly higher than that in normal group (P < 0.05). The relative expression level of E3RSIκB mRNA in the ulcerative collaterals group was significantly lower than that in the model group (P <0.05). The relative expression level of E3RSIκB protein in colonic mucosa in model group was significantly higher than that in normal group (P <0.05) The relative expression level of E3RSIκB protein in the group of high, medium and low dose of KD was significantly lower than that in the model group (P <0.05 or P <0.01). 【Conclusion】 UC can inhibit the expression of E3RSIκB, prevent the degradation of IκB ubiquitinated, and then inhibit the activation of NF-κB and reduce the inflammatory reaction.