康复新液对急性大鼠实验性结肠炎作用机制的研究

Study on the Mechanisms of Kangfuxin Solution in the Treatment of Acute Experimental Colitis in Rats

背景:康复新液在临床上对溃疡性结肠炎(UC)具有良好疗效,但其具体机制尚不明确。基质金属蛋白酶(MMPs)是细胞外基质(ECM)合成和降解的关键酶,与UC的发生、发展密切相关。目的:探讨康复新液对葡聚糖硫酸钠(DSS)诱导的急性大鼠实验性结肠炎的疗效和对MMPs表达的影响。方法:30只健康雄性Sprague-Dawley大鼠随机分为正常对照组、模型组和康复新液治疗组。模型组和康复新液治疗组大鼠以3%DSS溶液制备急性大鼠实验性结肠炎模型。第10d处死大鼠,测定各组大鼠疾病活动指数(DAI)、组织学损伤评分和肠组织髓过氧化物酶(MPO)活性;以MMPs功能基因芯片检测筛选表达差异的基因,并行荧光定量聚合酶链反应(PCR)验证。结果:与模型组相比,康复新液治疗组大鼠DAI和组织学损伤评分显著降低(P<0.01),MPO活性显著降低(P<0.01)。康复新液治疗组表达差异在2倍或以上的基因有16个,PCR结果显示MMP-3、MMP-13表达较模型组降低(P<0.01),与芯片变化趋势一致。结论:康复新液可改善DSS诱导的急性大鼠实验性结肠炎,减轻腹泻、血便症状。康复新液抑制MMP-3、MMP-13表达可能是其作用机制之一。

Kangfuxin solution has been demonstrated to ameliorate ulcerative colitis （UC） clinically, however, its mechanism remains unclear. Matrix metalloproteinases （MMPs） are the key enzymes in synthesis and degradation of extracellular matrix （ECM）, and are closely related to the development and progression of UC. Aims： To investigate the therapeutic effect of Kangfuxin solution on acute experimental colitis induced by dextran sulfate sodium （DSS） in rats and its effect on expression of MMPs. Methods： Thirty male Sprague-Dawley rats were randomly divided into three groups： normal control group, model group, and Kangfuxin solution-treated group. Acute experimental colitis model was established by 3% DSS in model group and Kangfuxin solution-treated group. All rats were sacrificed 10 days later. Disease activity index （DAD, score of histological injury and activity of intestinal tissue myeloperoxidase （MPO） were determined. Genes of differential expression were detected by MMPs related gene microarray, and the results were verified by fluorescence quantitative polymerase chain reaction （PCR）. Results： Compared with model group, DAI and score of histological injury in Kangfuxin solution-treated group decreased significantly （P〈0.01）, and activity of MPO decreased significantly （P〈0.01）. In comparison with model group, there were 16 genes changing 2-fold or more than 2-fold. PCR amplification showed that expressions of MMP-3 and MMP-13 in Kangfuxin solution-treated group decreased significantly （P〈0.01）, which was consistent with the changes seen in gene microarray. Conclusions： Kangfuxin solution may ameliorate acute experimental colitis induced by DSS in rats, and alleviate the diarrhea and bloody stool. Inhibition of MMP-3 and MMP-13 expressions may be one of the mechanisms of Kangfuxin solution.