Effect of Yu's Qingrehuoxue formula to ulcerative colitis rats'colon mucosa on the expression of NLRP3, IL-1β and NF-Κb
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摘要: 目的 研究俞氏清热活血方对溃疡性结肠炎(UC)大鼠结肠黏膜NOD样受体热蛋白结构域相关蛋白3(NLRP3)、白细胞介素-1β(IL-1β)和核转录因子-κB(NF-κB)表达的影响，以观察俞氏清热活血方减轻UC炎症反应的作用。方法 30只Wistar大鼠随机分为对照组、模型组、美沙拉嗪西药组、中药低、中、高剂量组(每组5只)，运用三硝基苯磺酸(TBSN)造模成功后，分别给予蒸馏水及高、中、低浓度俞氏清热活血方及美沙拉嗪混悬液灌肠14 d。运用蛋白质印迹法(WB)及免疫组化方法检测各组大鼠结肠黏膜NLRP3、IL-1β、NF-κB的表达。结果 WB及免疫组化结果均显示，与对照组比较，模型组大鼠结肠黏膜组织中NLRP3、IL-1β、NF-κB蛋白表达水平显著升高(P < 0.01)。与模型组比较，美沙拉嗪西药组及高、中、低剂量中药组大鼠结肠黏膜组织NLRP3、IL-1β、NF-κB蛋白表达水平显著降低(P < 0.05，P < 0.01)。结论 俞氏清热活血方可能是通过降低NLRP3、IL-1β和NF-κB的表达来减轻UC炎症反应，从而缓解UC临床症状。
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关键词:
- 俞氏清热活血方 /
- 溃疡性结肠炎 /
- 中药灌肠 /
- NOD样受体热蛋白结构域相关蛋白3
Abstract: Objective This study was done to study Yu's Qingrehuoxue formula's effect to ulcerative colitis rats'colon mucosa on the expression of NLRP3, IL-1β and NF-κB, to prove that Yu's formula can reduce inflammatory response.Methods Thirty Wistar rats were randomly divided into six groups, include control group, model group, mesalazine group, Chinese medicine high-dose, medium-dose and low-dose groups(5 in each group).After successful modeling with TNBS(trinitrobenzene sulfonic acid), the rats were given 14 days enema with distilled water, high, medium and low concentration Yu's Qingrehuoxue formula and mesalazine suspension. Use Western blot and immunohistochemistry to test the expression of NLRP3, IL-1β and NF-κB in colon mucosa in every group.Results Western blot and immunohistochemistry results all showed that compared with control group, the expression levels of NLRP3, IL-1β and NF-κB protein in colon tissue of model group were significantly increased(P < 0.01). Compared with model group, the expression levels of NLRP3, IL-1β and NF-κB protein in colon tissues of mesalazine, high-dose, medium-dose and low-dose groups were significantly decreased(P < 0.05 or P < 0.01).Conclusion Yu's Qingrehuoxue formula maybe through reducing the expression of NLRP3, IL-1β and NF-κB to reduce the inflammation in UC. -
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表 1 WB一抗具体信息
试剂 厂家 货号 稀释比 NLRP3/NALP3 Antibody Novus Biologicals NBP2-12446 1：1 000 IL-1β Antibody Santa Cruz sc-12742 1：200 NF-κB p65 Antibody CST 8242 1：1 000 
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表 2 免疫组化一抗具体信息
试剂 厂家 货号 稀释比 NLRP3/NALP3 Antibody Novus Biologicals NBP2-12446 1：30 IL-1β Antibody Santa Cruz sc-12742 1：200 NF-κB p65 Antibody CST 8242 1：500
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