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目的:探讨溃结灵对溃疡性结肠炎(UC)大鼠结肠黏膜外周血中性粒细胞(PMN)凋亡及Bcl-2基因表达的影响。方法:雄性SD大鼠90只完全随机分为正常对照组、模型组、艾迪莎阳性对照组及溃结灵高、中、低剂量组共6组,每组15只。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模型,第3天开始,除正常对照组外,各组分别按剂量给予生理盐水、艾迪莎、溃结灵处理,至7 d后处死动物。肉眼观察结合显微镜观察结肠组织形态学评分;采用流式细胞仪检测动物外周血PMN凋亡和Bcl-2基因表达水平。结果:中、高剂量溃结灵能显著改善大鼠UC及提高外周血PMN凋亡率(P<0.05);PMN凋亡率随炎症好转有上升趋势并与溃结灵剂量呈正相关。与模型组比较,溃结灵中、高剂量和艾迪莎均能降低Bcl-2基因表达水平(P<0.01),且溃结灵降低Bcl-2基因表达水平呈剂量相关性。PMN凋亡率与肉眼损伤评分、光镜损伤评分及Bcl-2表达水平相关系数分别为:-0.7957、-0.7681、-0.7527。结论:溃结灵对UC有明显治疗效果,并可能通过下调Bcl-2基因表达水平进而提高PMN凋亡率来发挥治疗作用。
Objective: To investigate the effect of Kuijieling on the apoptosis of peripheral neutrophils (PMN) and the expression of Bcl-2 in colon mucosa of ulcerative colitis (UC) rats. METHODS: Ninety male Sprague-Dawley rats were randomly divided into normal control group, model group, Aidisha positive control group and Kuijieling high-, middle-, and low-dose group. There were 6 groups, 15 in each group. The rat ulcerative colitis model was established with 5% 2,4,6-trinitrobenzene sulfonic acid 100 mg/kg enema. On the third day, except for the normal control group, each group was given doses of physiological saline, Eddy Sasuke and Kuei Jieling were treated and the animals were sacrificed 7 days later. The morphological scores of colon tissues were observed with naked eyes and microscopic observations. The apoptosis of PMNs and the expression of Bcl-2 mRNA in peripheral blood of animals were detected by flow cytometry. RESULTS: The high and medium doses of KUJI can significantly improve the UC of rats and increase the apoptosis rate of PMN in peripheral blood (P<0.05). The apoptosis rate of PMN increased with the increase of inflammation and was positively correlated with the dose of KUOJIULING. Compared with the model group, the expression of Bcl-2 gene was decreased in the group of Kuijieling, high dose, and Idesha (P<0.01), and the expression of Bcl-2 in Kuijieling was dose-dependent. The correlation coefficient between the apoptosis rate of PMN and the visual injury score, light microscope damage score, and Bcl-2 expression level were -0.7957, -0.7681, -0.7527, respectively. Conclusion: Kuijieling has obvious curative effect on UC, and may play a therapeutic role by down-regulating the expression of Bcl-2 gene and increasing the apoptotic rate of PMN.