固肠止泻丸对溃疡性结肠炎小鼠的治疗作用研究

Therapeutic effects of Guchang Zhixie Pills on mice with ulcerative colitis

目的研究固肠止泻丸(Guchang Zhixie Wan,GC)对葡聚糖硫酸钠(destran sulfate sodium,DSS)诱导的溃疡性结肠炎(ulcerative colitis,UC)的治疗作用及其作用机制。方法36只雄性小鼠随机均分为对照组、模型组、柳氮磺胺吡啶组、GC组,除对照组外,其余3组小鼠自饮DSS进行UC造模。待UC造模成功后,模型组与对照组灌胃相应体积纯净水,柳氮磺胺吡啶组(125 mg/kg)和GC组(50 mg/kg)灌胃相应药物,连续灌胃观察5 d。给药前后观察小鼠体征,并计算小鼠疾病活动指数(disease activity index,DAI);HE染色观察结肠组织病理学变化;ELISA法检测血清炎症因子白细胞介素-4(interleukin-4,IL-4)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-8(interleukin-8,IL-8)、白细胞介素-10(interleukin-10,IL-10)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量;Western blot法检测小鼠结肠组织炎症相关信号蛋白核因子κB(nuclear factor-κB,NF-κB)、核因子κB抑制蛋白α(inhibitor alpha of NF-κB,IκBα)、信号转导及转录激活因子3(signal transduction and activator of transcription 3,STAT3)和磷酸化信号转导及转录激活因子3(phosphorylated signal transduction and activator of transcription 3,p-STAT3)的表达。结果与模型组比较,柳氮磺胺吡啶组和GC组DAI评分均下降(P<0.01);模型组可见炎性异常浸润;柳氮磺胺吡啶组和GC组结肠组织损伤相对恢复;与对照组比较,模型组IL-6、IL-8及TNF-α含量升高(P<0.01),IL-4、IL-10含量降低(P<0.05);与模型组比较,GC组IL-6、IL-8及TNF-α含量下降(P<0.05),IL-4含量升高(P<0.01);与柳氮磺胺吡啶组比较,GC组IL-10表达降低(P<0.05);与对照组比较,模型组中NF-κB和IκBα蛋白表达降低(P<0.01),p-STAT3水平升高(P<0.05);与模型组比较,GC组NF-κB和IκBα蛋白表达升高(P<0.05),柳氮磺胺吡啶组和GC组p-STAT3水平降低(P<0.05);与柳氮磺胺吡啶组比较,GC组NF-κB蛋白表达升高(P<0.05)。结论GC对DSS诱导的UC小鼠具有显著的治疗作用,且这一作用可能与NF-κB和STAT3信号通路相关。

Objective To explore the therapeutic effects and mechanisms of Guchang Zhixie Pills(GC)on ulcerative colitis(UC)induced by dextran sulfate sodium(DSS).Methods A total of 36 male mice were randomly divided into control group,model group,sulfasalazine group,and GC group.Except the control group,the other three groups of mice were induced by DSS.After the UC model was successfully established,the mice in control and model group were given the corresponding volume of pure water by intragastric administration.The sulfasalazine group and GC group were administered with 125 mg/kg of sulfasalazine and 50 mg/kg of GC respectively for 5 d.Physical signs of the mice were observed,and the disease activity index(DAI)was calculated before and after administration.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of colon tissues in each group.The serum levels of interleukin-4(IL-4),interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10(IL-10),and tumor necrosis factor-α(TNF-α)were detected by ELISA.We also tested the expressions of nuclear factor-κB(NF-κB),inhibitor alpha of NF-κB(IκBα),signal transduction and activator of transcription 3(STAT3)and phosphorylated signal transduction and activator of transcription 3(p-STAT3)in colon tissues of mice in each group by Western blot.Results The DAI scores of the sulfasalazine group and the GC group decreased compared with the model group(P<0.01);the colon tissue of the model group had abnormal inflammatory infiltration;the pathological damage of the colon tissues in the sulfasalazine group and GC group recovered relatively;compared with the control group,the levels of IL-6,IL-8 and TNF-αin the model group increased(P<0.01),and the levels of IL-4 and IL-10 decreased(P<0.05);compared with model group,the levels of IL-6,IL-8 and TNF-αin the GC group decreased(P<0.05),and the levels of IL-4 increased(P<0.01);compared with sulfasalazine group,the levels of IL-10 in GC group decreased(P<0.05);compared with control group,the expression of NF-κB and IκBαprotein of the model group decreased(P<0.01),and the expression of p-STAT3 protein increased(P<0.05);compared with model group,the expression of NF-κB and IκBαprotein in GC group increased(P<0.05),and the expression of p-STAT3 protein in sulfasalazine group and GC group decreased(P<0.05);compared with the sulfasalazine group,the expression of NF-κB protein of the GC group increased(P<0.05).Conclusion GC has the obvious therapeutical effects on DSS-induced UC in mice and the therapeutic effects may be related to the NF-κB and STAT3 pathways.