溃结灵对溃疡性结肠炎大鼠结肠黏膜的CINC-1及其受体CXCR2表达的作用

The effect of Kuijieling decoction on the expression of cytokine-induced neutrophil chemoattractant-1 and CXC chemokine receptor 2 in a rat model of colitis

目的探讨溃结灵对溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠黏膜的中性粒细胞趋化因子-1(cytokine-induced neutrophil chemoattractant-1,CINC-1)及其受体CXC趋化因子受体2(CXC chemokine receptor 2,CXCR2)表达的作用。方法90只雄性SD大鼠随机分为正常组、模型组、艾迪莎组、溃结灵高剂量组、溃结灵中剂量组和溃结灵低剂量组,共6组,每组15只。用5%2,4,6-三硝基苯璜酸(2,4,6-trinitrobenzene sulfonic acid,TNBs)灌肠建立大鼠溃疡性结肠炎模型。造模成功后,除正常组外,各组分别给予生理盐水0.33 mL、艾迪莎0.125g.kg-1.d-1、溃结灵高(18.3 g.kg-1.d-1)、中(9.2 g.kg-1.d-1)、低(4.6 g.kg-1.d-1)剂量处理,7 d后处死动物。肉眼观察结合显微镜观察对结肠组织形态学评分;运用荧光定量PCR法及免疫组化染色法分别检测各组大鼠CINC-1及CXCR2 mRNA及其蛋白表达。结果溃结灵具有明确改善大鼠UC的效应,与艾迪莎组相比,溃结灵高剂量组的疗效差异无统计学意义(P>0.05),中、低剂量组的疗效差异有统计学意义(P<0.01);与正常组相比,模型组CINC-1及CXCR2 mRNA及蛋白质表达呈明显上升趋势(P<0.05),与炎症呈正相关;经过溃结灵和艾迪莎干预后,随着炎症改善,CINC-1及CXCR2 mRNA及其蛋白表达呈下降趋势。结论溃结灵可能通过干预CINC-1及CXCR2的表达来抑制中性粒细胞趋化,从而达到缓解TNBs诱导溃疡性结肠炎的效应。

Objective To detect the effect of Kuijieling decoction（KD） on cytokine-induced neutrophil chemoattractant-1（CINC-1） and CXC chemokine receptor 2（CXCR2） in a rat model of colitis. Methods Ninty SD rats were randomly divided into 6 groups with 15 rats in each group： normal group,blank group,etiasa solution group,and 3 KD groups at the doses of 18.3,9.2 and 4.6 g·kg^-1·d^-1,respectively.The rat model of ulcerative colitis（UC） was brought by an enema of 100 mg/kg of 5% 2,4,6-trinitrobenzene sulfonic acid（TNBs） and 50% ethanol.From the 3rd day,except for normal group,other groups were treated with an enema of 0.9% NaCl 0.33 mL,etiasa 0.125 g·kg^-1·d^-1,and KD,respectively.The rats were sacrificed 1 week later.The histomorphology changes of the colon were evaluated by naked eyes and microscope.The expression of CINC-1 and CXCR2 were detected with the technique of immunohistochemical staining and real time-PCR. Results There was no statistical difference between etiasa solution and KD（18.3 g·kg^-1·d^-1） in therapeutic effect on UC in the rat model（P〈0.05）,but the difference between KD groups（9.2 and 4.6 g·kg^-1·d^-1） and etiasa solution group had statistical significance（P〈0.01）.Compared with normal group,the expression of CINC-1 and CXCR2 in blank group increased（P〈0.05）.These changes showed a positive correlation with inflammation in the rat model of colitis.After treatment with etiasa solution or KD,the expression of CINC-1 and CXCR2 decreased. Conclusions These data demonstrate that KD is shown to exert treatment effect on the rat model of UC,which was possibly achieved by reducing the expression of CINC-1 and CXCR2 and inhibiting infiltration of polymorphnuclear neutrophil.