黄连粗多糖协同小檗碱改善溃疡性结肠炎肠黏膜屏障损伤的作用

Coptidis Rhizoma Crude Polysaccharide and Berberine Synergistically Restore Intestinal Mucosal Barrier Damage in Ulcerative Colitis

目的:考察黄连粗多糖与小檗碱在溃疡性结肠炎小鼠治疗中的协同作用。方法:采用雄性BALB/c小鼠30只随机分为5组,除正常组6只外,其余在小鼠日常饮水中给予5%葡聚糖硫酸钠建立结肠炎模型。建模成功后,每组分别进行灌胃给药4 d,每日1次,小檗碱组(100 mg·kg^(-1)BBR)、小檗碱+低剂量粗多糖组(100 mg·kg^(-1)BBR+22.8 mg·kg^(-1)CCP)、小檗碱+高剂量粗多糖组(100 mg·kg^(-1)BBR+45.6 mg·kg^(-1)CCP),模型组和正常组灌胃同体积生理盐水。实验结束后处死小鼠提取结肠,通过蛋白免疫印迹法检测小鼠结肠组织紧密连接闭锁连接蛋白-1(ZO-1)、紧密连接蛋白-1(Claudin-1)和闭合蛋白(Occludin)蛋白的表达。以正常组为对照,评价各治疗组模型小鼠疾病活动指数(DAI)评分、结肠长度、结肠组织形态和结肠紧密连接蛋白的表达水平。结果:与正常组比较,模型组小鼠ZO-1、Claudin-1和Occludin蛋白表达显著降低(P<0.01);与模型组比较,小檗碱组Claudin-1蛋白表达无明显差异,ZO-1和Occludin蛋白表达显著升高(P<0.01),黄连粗多糖联合小檗碱组Claudin-1、ZO-1和Occludin蛋白表达显著升高(P<0.01);与小檗碱组比较,黄连粗多糖联合小檗碱组紧密连接蛋白表达明显强于小檗碱单独给药(P<0.05)。结论:黄连粗多糖协同小檗碱给药能有效改善溃疡性结肠炎小鼠肠黏膜屏障损伤。

Objective:To investigate the synergistic effect of Coptidis Rhizoma crude polysaccharide(CCP)and berberine(BBR)in treating ulcerative colitis(UC)model mice.Method:Thirty male BALB/c mice were randomized into five groups.Except the 6 mice in the normal group,the rest were given 5%dextran sodium sulfate in their daily drinking water to establish the UC model.After modeling,the mice were administrated with corresponding agents by gavage once daily for 4 days:BBR(100 mg·kg^(-1))group,BBR(100 mg·kg^(-1))+low-dose(22.8 mg·kg^(-1))CCP group,BBR(100 mg·kg^(-1))+high-dose(45.6 mg·kg^(-1))CCP group.The mice in the model group and normal group were administrated with the same volume of normal saline.At the end of the experiment,the mice were sacrificed for the collection of colon,and the expression of tight junction proteins zonula occluden-1(ZO-1),Claudin-1,and Occludin in colon tissue was detected by Western blot.With the normal group as the control,the disease activity index(DAI)score,colon length,colon histomorphology,and expression levels of tight junction proteins in other groups were evaluated.Result:Compared with the normal group,the modeling down-regulated the protein levels of ZO-1,Claudin-1,and Occludin(P<0.01).Compared with the model group,BBR did not significantly change the protein level of Claudin-1 and up-regulated those of ZO-1 and occludin(P<0.01).The expression levels of Claudin-1,ZO-1,and Occludin were up-regulated in BBR+CCP groups(P<0.01).The expression levels of tight junction proteins in BBR+CCP groups were significantly higher than those in the BBR group(P<0.05).Conclusion:The administration of CCP combined with BBR can effectively ameliorate intestinal mucosal barrier damage in the mice with UC.