基于尤斯室技术研究黄连水煎液中微粒体系对小檗碱肠黏膜透过特性的影响及相关机制

Effect and Mechanism of Particle Aggregates in Coptis chinensis Decoction on the Intestinal Mucosal Permeability of Berberine Based on Ussing Chamber Technique

目的采用尤斯室(Ussing Chamber)技术考察黄连水煎液中微粒体系对小檗碱(berberine,Ber)离体肠黏膜透过特性的影响及相关机制研究。方法常规煎煮黄连,高速离心得黄连水煎液中的微粒体系;采用尤斯室技术,考察Ber在大鼠不同肠段的吸收情况以及黄连水煎液中微粒体系对Ber肠黏膜透过的影响;采用Western blot检测肠黏膜中紧密连接相关蛋白ZO-1、Occludin、claudin-1的蛋白表达水平。结果 Ber在大鼠小肠全肠段均有吸收,空肠吸收最好;黄连水煎液中的微粒体系显著促进Ber的吸收;微粒体系可显著下调小肠上皮细胞紧密连接相关蛋白ZO-1与Occludin的表达水平,且与加入微粒体系的剂量呈正相关。结论基于尤斯室技术建立动物离体实验模型,黄连水煎液中微粒体系能显著增加Ber的肠黏膜透过性,这可能与其下调紧密连接蛋白表达有关。

OBJECTIVE To investigate the effect and mechanism of particle aggregates in Coptis chinensis decoction on the permeability of berberine(Ber) through isolated intestinal mucosa based on Ussing chamber technique. METHODS Coptis chinensis decoction was prepared by conventional decocting method, and the particle aggregates of Coptis chinensis decoction were obtained by high-speed centrifugation. The absorption of Ber in different intestinal segments of rats and the effect of particle aggregates in Coptis chinensis decoction on the permeability of Ber intestinal mucosa were studied by Ussing chamber technique. The protein expression levels of ZO-1, occludin and claudin-1 in intestinal mucosa were detected by Western blot. RESULTS Ber was absorbed in the whole intestinal segment of rats, and the jejunum was the best. Particle aggregates in Coptis chinensis decoction significantly promoted the absorption of Ber. Particle aggregates could significantly down-regulate the expression level of ZO-1 and occludin, which was positively correlated with the dosage of solid particles. CONCLUSION The animal model in vitro was established by Ussing chamber technique, the particle aggregates in Coptis chinensis decoction can significantly increase the intestinal permeability of Ber, which may be related to the down-regulation of the expression of tight junction protein.