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溃结灵对溃疡性结肠炎大鼠中性粒细胞凋亡的影响及其机制

Effect and mechanism of Kuijieling decoction on the apoptosis of polymorph nuclear neutrophil in the model of ulcerative colitis

  • 摘要: 探讨溃结灵对溃疡性结肠炎大鼠结肠黏膜外周血中性粒细胞(PMN)的作用及其机制。采用5% 2,4,6三硝基苯磺酸100 mg/kg灌肠建立大鼠溃疡性结肠炎(UC)模型,动物分为模型组、治疗组(高、中、低剂量)和阳性对照组。第3天开始,除正常组外,各组分别按剂量给予生理盐水、美沙拉嗪(阳性对照)、溃结灵处理,至第10天后处死动物。肉眼结合显微镜观察结肠组织,进行形态学评分;采用流式细胞仪检测动物外周血PMN凋亡,ELISA法检测 IL-8、NO、ICAM-1水平。结果发现,中、高剂量(每日9.2、18.3 g/kg)溃结灵具有改善大鼠UC和提高外周血PMN凋亡率的效果(P<0.05);凋亡率随炎症好转有上升趋势并与溃结灵的使用剂量呈正相关;与模型组比较,溃结灵中、高剂量组和美沙拉嗪组均显著降低IL-8、NO和ICAM-1的水平(P<0.01),且IL-8、NO和ICAM-1水平降低与溃结灵剂量呈正相关;PMN凋亡率与IL-8、NO和ICAM-1相关系数分别为:-0.792 6、-0.705 1、-0.593 7。结果提示溃结灵对UC有明显治疗效果,并可能通过下调IL-8、NO、ICAM-1水平进而提高PMN凋亡率来发挥治疗作用。

     

    Abstract: To investigate the effect and mechanism of Kuijieling decoction(KD) on the apoptosis of polymorph nuclear neutrophil (PMN) in the model of ulcerative colitis,the rat model of ulcerative colitis was induced by 100 mg/kg of 2,4,6-trinitrobenzene sulfonic acid (TNBS) enema.The experimental animals were randomly divided into Kuijieling treatment group (including high,medium and low dosage),model control group,and mesalazine positive control group.Each group, except for the normal group,was treated from 3rd day with an enema of 0.9% saline、mesalazine and KD respectively.The rats were sacrificed 10 days later.The histomorphological changes of the colon were evaluated by naked eyes and microscope.The apoptosis of PMN was tested by flow cytometry and the content of IL-8,NO and ICAM-1 was detected with ELISA in the peripheral blood of rats.There were significant effects on ulcerative colitis and in increasing the apoptosis of PMN in the KD (9.2 and 18.3 g·kg-1·d-1) groups and mesalazine solution group (P<0.05).The apoptosis were positively correlated with inflammation in the model of colitis in a dose-dependent relationship with KD.Compared with the model group,the contents of IL-8,NO and ICAM-1 were decreased in the groups of KD (9.2 and 18.3·kg-1·d-1) and mesalazine solution (P<0.01).The correlation coefficients between the apoptosis and IL-8,NO,ICAM-1 were -0.792 6,-0.705 1 and -0.593 7 respectively.The results demonstrated that Kuijieling decoction exerted a therapeutic effect on the rat model of ulcerative colitis possibly by reducing the content of IL-8,NO and ICAM-1 to increase the apoptosis of PMN.

     

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