黄连提取物小檗碱对溃疡性结肠炎小鼠结肠黏膜机械屏障的影响

Effects of Berberine extracted from Chinese Goldthread Rhizome on the intestinal mucosal mechanical barrier in mice with ulcerative colitis

目的观察黄连提取物小檗碱(BBR)对溃疡性结肠炎(UC)小鼠结肠黏膜机械屏障的干预作用,探讨BBR治疗UC的可能机制。方法 BALB/c小鼠随机分为模型对照组、BBR低、高剂量组、阳性对照组和空白对照组。采用右旋葡聚糖硫酸复制UC模型,灌胃给药7 d,评估疾病活动指数(DAI),结肠组织切片、HE染色并观察组织的病理学变化;免疫组化SP法检测紧密连接蛋白claudin-1、occludin和ZO-1的表达水平;Western Blot法测定肠道干细胞(ISCs)标志物LGR-5和TERT的含量。结果与治疗前比较,药物治疗后小鼠的DAI评分均下降(P<0.01),结肠组织病理表现有不同程度的改善;与治疗后同期模型对照组比较,高剂量BBR组的DAI评分显著下降(P<0.01),结肠组织中claudin-1、occludin和ZO-1的水平均明显升高(P<0.01)。BBR低、高剂量组结肠组织中LGR-5和TERT的含量均升高(P<0.05)。结论 BBR通过抑制ISCs和紧密连接蛋白的破坏,维持肠黏膜机械屏障的稳态,从而有效抑制UC小鼠的结肠炎症,对UC的发生发展起到保护作用。

Objective To observe the effects of Berberine （BBR） extracted from Chinese Goldthread Rhizome on colonic mucosal mechanical barrier in mice with ulcerative colitis （ UC）, and to explore the possible mechanisms of BBR＇ s ther- apeutic effect. Methods BALB/c mice were randomly divided into five groups ： model control group, low and high dose BBR group, positive control group and blank control group. UC model was established by dextran sulphate sodium, and daily garage administration was given for 7 days. Disease activity index （DAI） of mice were assessed, colon tissue were sectioned and HE stained, and its pathological changes were observed, the contents of claudin-1, occludin and ZO-1 were detected by immunohistochemical method, the expression levels of LGR-5 and TERT were determined by Western Blot. Results Compared with those before the treatment, the DAI of UC mice after the treatment were all decreased （P 〈 0.01 ）, with their pathological symptoms of colitis improving to varying degrees; compared with model control group after the treatment, the DAI of high dose BBR group were decreased obviously （P 〈 0.01 ）, the expression levels of claudin-1, occludin and ZO-I in high dose BBR group were all markedly increased （ P 〈 0.01 ）, and the contents of LGR-5 and TERT in low and high dose BBR groups were all elevated （ P 〈 0.05）. Conclusion Berberine can keep the steady state of intestinal mucosal barrier by suppressing the destruction of ISCs and tight junction proteins, thus effectively inhibit the colitis in UC mice. It is then proved to have a protective effect on the pathogenesis of UC.