摘要
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 explore the effect of Kuijingling decoction（ KD） on NLRP3 inflamasomes regulatory in clonic mucosal of Rats with ulcerative colitis（ UC）,and to reveal the therapeutic mechanism of KD for UC. Methods： TNBS was used for the establishment of UC rat model,Sixty Rats were randomly divided into the six group： normal control（ NC） group,model control（ MC） group,Kuijieling high dose（ KHD）group,middle dose（ KMD）,low dose（ KLD）,and SASP group. After treatment,the rats were killed to get their colonic tissue. Expressions of NLRP3,caspase-1and ASC were detected by Western blotting and RT-q PCR. IL-18,IL-33 were detected by Elisa. Results： the model group＇ s pathological examination showed colon epithelial erosion,bleeding,multifocal ulcers and numerous inflammation cells infiltrating.However,the symptoms were improved in other groups. Compared with the normal group,The expression of Nlrp3,Asc,Caspase-1 protein and mRNA in the model group was obviously higher than that in the normal group（ P〈0. 01 OR 0. 05）; The expressions of Nlrp3、Asc、aspase-1 protein and mRNA in the KD high dose group（ 18. 3g/kg）,KD middle dose group（ 9. 2g/kg） 、KD low dose group（ 4. 6g/kg） and SASP group（ 0. 5g/kg） were lower than those in model groups（ P〈0. 01 OR 0. 05）; the model group＇ s IL-18,IL-33 expressions was increase.KD high dose group（ 18. 3g/kg）,KD middle dose group（ 9. 2g/kg） 、KD low dose group（ 4. 6g/kg） and SASP group（ 0. 5g/kg） was decrease. Conclusion： ： The mechanism of Kuijingling prevent the ulcerative colitis in Rats may be related to the inhibition of NLRP3 inflamasomes protein and mRNA expressions and decrease the IL-18,IL-33 expressions.
作者
覃景春
钱彩云
揭凤鸣
甘丽萍
李燕舞
巫燕莉
杜群
Qin Jingchu;Qian Caiyun;Jie Fengming;Gan Liping;Li Yanwu;Wu Yanli;Du Qun(Pi Wei Institution,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405)
出处
《中药药理与临床》
CSCD
北大核心
2017年第4期90-94,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金面上项目(81373798)
