小檗碱对酒精性肝炎小鼠“肠漏”的保护作用*

doi:10.3969/j.issn.1672-5069.2017.06.007

摘要/Abstract

摘要： 目的探讨小檗碱（BBR）对酒精性肝炎小鼠“肠漏”的保护作用。方法将40只雄性C57BL/6小鼠随机分为对照组、酒精组、BBR组和酒精联合BBR组,饲养8 w。采用ELISA法检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和内毒素（LPS）水平。常规行组织病理学和透射电镜检查肝组织学改变和肠上皮细胞间紧密连接（TJ）超微结构。体外培养肠上皮细胞Caco-2,采用免疫荧光法检测经酒精和BBR处理后的细胞间紧密连接主要结构蛋白occludin表达。结果酒精处理组小鼠血清ALT、AST和LPS水平分别为（46.5± 5.9） U/L、（57.0± 6.1） U/L和（0.40± 0.05） ng/ml,显著高于对照组（P< 0.01）;酒精联合BBR组血清ALT、AST和LPS水平分别为（27.6± 4.2） U/L、（31.8± 4.1） U/L和（0.24± 0.03） ng/ml,均显著低于酒精组（P< 0.05）,而与对照组无显著性差异（P> 0.05）;酒精联合BBR组肝组织细胞变性和炎细胞浸润较酒精干预组明显改善;电镜下,可见酒精处理组小鼠肠上皮细胞间紧密连接结构模糊、缝隙明显增宽,酒精联合BBR组以上改变明显减轻;酒精处理组Caco-2细胞膜occludin分布减少、中断,部分进入胞浆内,酒精联合BBR组分布虽也减少,但仍连续,无中断。结论小檗碱能改善慢性乙醇摄入诱导的细胞膜occludin减少和上皮细胞间紧密连接结构破坏,改善“肠漏”而具有保护肝脏作用。*基金项目：北京市自然科学基金资助项目(编号：7154194）;国家自然科学基金资助项目（编号：81570536）作者单位：150086 哈尔滨市哈尔滨医科大学附属第二医院体检中心（王洪岩）;北京市首都医科大学附属北京天坛医院消化内科（李鑫,迟程,徐有青）第一作者：王洪岩,女,31岁,医学博士,住院医师。主要从事酒精性肝病的防治研究。E-mail: xinglin_freedom@126.com

关键词: 酒精性肝炎, 小檗碱, 肠漏, 紧密连接, Occludin蛋白, 小鼠

Abstract: 【Abstrat】 Objective To explore the protective effect of berberine on intestinal leak in mice with alcoholic hepatitis. Methods Forty male C57BL/6mice were randomly divided into control, alcohol, berberine, alcohol plus berberine group with10animals in each. Mice were fed with alcohol or vehicle for8weeks, all were sacrificed. Serum alanine aminotransferas （ALT）, aspartate aminotransferase（AST）, lipopolysaccharide （LPS） were detected by enzyme-linked immunosorbent assay（ELISA）. The pathologic changes of liver tissues were assessed by HE staining, and the ultrastructures of tight junction（TJ） complex was observed by transmission electron microscope. In vitro, occludin, the main structural protein of intercellular TJ, was detected by immunofluorescence in intestinal epithelial Caco-2cells, which were cultured with ethanol （EtOH）, BBR, EtOH plus BBR respectively. Results Serum ALT, AST, LPS levels in alcohol group were（46.5±5.9） U/L, （57.0±6.1） U/L, （0.40±0.05） ng/ml, respectively, significantly higher than those in control group （P<0.01）;There was no significant difference between alcohol plus BBR group, control group;Serum ALT, AST, LPS levels in alcohol plus BBR group were （27.6±4.2） U/L, （31.8±4.1） U/L and（0.24±0.03） ng/ml, respectively, significantly lower than those in alcohol group（P<0.05）;The histopathologic examination of liver tissues showed alcoholic hepatitis in alcohol group, and it was alleviated as respect to steatosis, inflammatory cell infiltration in alcohol plus BBR group;Electron micrographs showed that the TJ structures between intestinal epithelial cells were fuzzy, the gaps were expanded obviously in alcohol group, and all of these were improved obviously in alcohol plus BBR group;Immunofluorescence indicated that the occludin on Caco-2cell membranes were decreased, interrupted, partly entered into cytoplasm in alcohol group, while in alcohol plus BBR group, the distribution of occludin was decreased, though, but was continuous.Conclusion Berberine improves the decrease in occludin expression on cell membranes, the disruption of TJ structures between intestinal epithelial cells induced by chronic ethanol intake, alleviates intestinal leak

Key words: Alcoholic hepatitis, Berberine, Intestinal leak, Tight junction, Occludin, Mice

王洪岩, 迟程, 徐有青, 李鑫. 小檗碱对酒精性肝炎小鼠“肠漏”的保护作用*[J]. 实用肝脏病杂志, 2017, 20(6): 663-667.

Wang Hongyan,Chi Cheng,Xu Youqing. Protective effect of berberine on intestinal leak in mice with alcoholic hepatitis[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2017, 20(6): 663-667.

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