论文部分内容阅读
目的:观察三硝基苯磺酸(TNBS)溃疡性结肠炎大鼠病理形态、结肠黏膜NLRP3炎性体蛋白表达、mRNA及其相关因子IL-18、IL-33的表达,从而揭示其可能的作用机制。方法:采用三硝基苯磺酸(TNBS)复制溃疡性结肠炎模型,60只大鼠随机分为6组,即正常组,模型组,柳氮磺砒啶(SASP)阳性对照组,溃结灵高、中、低剂量组。连续治疗10天后,采集粘膜样本。观察大鼠病理形态学变化,采用蛋白质印迹法和荧光定量PCR检测大鼠结肠组织NLRP3、ASC、caspase-1蛋白和mRNA的表达;ELISA法检测大鼠结肠粘膜中IL-18、IL-33的含量。结果:模型组大鼠出现不同程度的体重减轻、结肠缩短和结肠质量增加,病理结果显示固有层崩解,出血、溃疡形成,毛细血管扩张、充血,肌层及外膜内大量急慢性炎症细胞浸润。溃结灵高(18.3 g/kg)中(9.2g/kg)低(4.6g/kg)剂量组和柳氮磺砒啶组(0.5 g/kg)上述情况较模型组明显减轻。与正常组比较,模型组NLRP3、caspase-1、ASC的蛋白和mRNA表达显著增高;与模型组比较溃结灵高(18.3 g/kg)中(9.2g/kg)低(4.6 g/kg)剂量组、柳氮磺砒啶组(0.5 g/kg)NLRP3、caspase-1、ASC mRNA表达均显著减少,溃结灵高(18.3 g/kg)中(9.2g/kg)剂量组NLRP3、caspase-1、ASC的蛋白表达均显著减少,溃结灵低(4.6 g/kg)剂量组NLRP3、caspase-1、ASC的蛋白表达有下降趋势但无统计学意义;与正常组比较,模型组IL-18、IL-33的表达水平明显升高,溃结灵高(18.3 g/kg)中(9.2g/kg)低(4.6 g/kg)剂量组和柳氮磺砒啶组(0.5 g/kg)IL-18、IL-33的含量明显低于模型组。结论:溃结灵治疗UC的作用机制可能与抑制NLRP3炎性体各组分蛋白和mRNA表达的表达,并下调结肠黏膜IL18、IL33的含量,使得炎症缓解从而发挥治疗溃疡性结肠炎的作用。
OBJECTIVE: To observe the pathomorphology of TNBS-induced ulcerative colitis and the expression of NLRP3 inflammatory protein in colonic mucosa and the mRNA and its related factors IL-18 and IL-33, so as to reveal their possible Mechanism. Methods: The model of ulcerative colitis was replicated by trinitrobenzene sulfonic acid (TNBS). Sixty rats were randomly divided into six groups: normal group, model group, sulfasalazine (SASP) positive control group, Ling high, medium and low dose group. Mucosal samples were collected after 10 days of continuous treatment. The pathological changes of rats were observed. The protein and mRNA expressions of NLRP3, ASC and caspase-1 were detected by Western blot and real-time PCR. The levels of IL-18 and IL-33 in colonic mucosa content. Results: The rats in the model group showed different degrees of weight loss, shortened colon and colonic mass. The pathological findings showed that the lamina propria was disintegrated, hemorrhage, ulcer formation, telangiectasia, hyperemia and a large number of acute and chronic inflammatory cells infiltration. Compared with the model group, the above-mentioned conditions in the dose of 4.6g / kg and the group of sulfasalazine (0. 5g / kg) in the group of KFD (18.3 g / kg) The protein and mRNA expressions of NLRP3, caspase-1 and ASC in the model group were significantly increased compared with those in the normal group (4.6 g / kg) in the ulcer group (18.3 g / kg) Dose group, the expressions of NLRP3, caspase-1 and ASC mRNA in sulfasalazine group (0.5 g / kg) were significantly decreased. The levels of NLRP3, caspase The protein expression of NLRP3, caspase-1 and ASC in decoction group (4.6 g / kg) had a downward trend but no statistical significance. Compared with normal group, the expression of IL-1 and ASC protein in model group was significantly decreased The expression of IL-33 and IL-33 were significantly increased in the group of low dose (9.2 g / kg) of kympathoderma (18.3 g / kg) and the dose of 0.5 g / kg) IL-18, IL-33 levels were significantly lower than the model group. Conclusion: The mechanism of Kuijieling in treating UC may be related to inhibiting the expression of protein and mRNA in various components of NLRP3 inflammasome, decreasing the content of IL18 and IL33 in colonic mucosa, and relieving the inflammation and thus treating ulcerative colitis.