摘要
目的基于肠道M细胞模型,研究黄连多糖对小檗碱经小肠吸收的影响及相关机制,为研究中药多糖类成分影响活性成分吸收机制奠定基础。方法以Caco-2细胞与淋巴细胞分别构建小肠Caco-2细胞单层膜模型与小肠M细胞模型;考察不同浓度黄连多糖溶液(浓度分别为20、40和80μg·mL^-1)分别对小檗碱经两种小肠细胞模型转运的影响;以荧光素经小肠细胞模型吸收的Papp值反映黄连多糖对小肠细胞通透性的影响;以Western blot法检测紧密连接蛋白ZO-1的表达水平,分析黄连多糖对小肠细胞模型完整性的影响。结果成功构建小肠Caco-2细胞单层膜模型与小肠M细胞模型;在小肠M细胞模型中,黄连多糖溶液浓度与小檗碱转运率和细胞膜通透性成正相关、与紧密连接蛋白ZO-1表达水平成负相关。当黄连多糖溶液总糖浓度为80μg·mL^-1时,小檗碱转运率、细胞膜通透性以及紧密连接蛋白ZO-1表达水平与空白对照组间差异均有统计学意义(P<0.05或P<0.001);在小肠Caco-2细胞单层膜模型中,黄连多糖对小檗碱的转运率、细胞膜通透性以及紧密连接蛋白ZO-1的表达均没有显著影响(P>0.05)。结论黄连多糖可通过增加细胞膜通透性和降低紧密连接蛋白ZO-1的表达来促进小檗碱经小肠M细胞模型的转运,推测这可能与黄连多糖经M细胞摄取后对小肠相关免疫的调节作用有关。
Objective To determine the effect of Coptis chinensis polysaccharides on the intestinal berberine absorption and related mechanism,based on the M cell model,and to elucidate the effect of polysaccharides on the absorption of active ingredients in traditional Chinese medicine.Methods Caco-2 cells and lymphocytes were used to establish Caco-2 monolayer cell model and M cell model.Coptis Chinensis polysaccharide solutions at 20,40,and 80μg·mL^-1 were used to determine the effect of Coptis Chinensis polysaccharides on berberine transport through the two cell models.Papp value of fluorescein transport and expression level of tight junction protein ZO-1 were respectively analyzed to measure the effect of Coptis chinensis polysaccharides on the permeability and integrity of the two cell models.Results Caco-2 monolayer cell model and M cell model were successfully established.In the M cell model,Coptis chinensis polysaccharides exhibited concentrationdepend promotion in berberine transport and cell membrane permeability,and inhibition on ZO-1 expression level,especially Coptis chinensis polysaccharide solution at 80μg·mL^-1,which exhibited significantly higher berberine transport,higher cell membrane permeability and lower ZO-1 expression level than those of the control group(P<0.05 or P<0.001).In the Caco-2 monolayer cell model,Coptis chinensis polysaccharides,no matter at what concentration,did not significant affect the berberine transport,cell membrane permeability and expression of ZO-1(P>0.05).Conclusion Coptis chinensis polysaccharides can promote the transport of berberine through the M cell model by increasing the cell membrane permeability and decreasing the expression of tight junction protein ZO-1.This may be related to the regulation of Coptis chinensis polysaccharides on the small intestine-related immunity through the phagocytosis/uptaking by M cells.
作者
吴晶晶
袁秀妍
杨晔
WU Jing-jing;YUAN Xiu-yan;YANG Ye(School of Pharmacy,Anhui University of Traditional Chinese Medicine,Hefei 230012;Institute of Pharmaceutical Preparation,Anhui Academy of Traditional Chinese Medicine,Hefei 230012;Key Laboratory of Chinese Medicine Compound of Anhui,Hefei 230012)
出处
《中南药学》
CAS
2020年第4期525-530,共6页
Central South Pharmacy
基金
国家自然科学基金资助项目(No.81874348,No.81303239)
安徽省自然科学基金(No.1908085J29)
安徽省重点研究与开发计划(对外科技合作)专项(No.201904b11020023)
安徽省教育厅高校自然科学研究重点项目(No.KJ2018A0282)。
关键词
黄连多糖
小檗碱
Caco-2细胞单层膜模型
M细胞模型
跨膜转运
Coptis chinensis polysaccharide
berberine
the Caco-2 monolayer cell model
M cell model
transmembrane transport
