白术多糖对小肠上皮细胞细胞屏障及黏附连接蛋白表达的影响  

Effect of Atractylodes macrocephala Polysaccharides on IEC-6 Cell Barrier and Expression of Adhesion Junction Proteins

作　　者：罗蒙 伍婷婷[1] 李茹柳[1] 张东 朱惠彬 胡玲[1] 陈蔚文[1] LUO Meng;WU Tingting;LI Ruliu;ZHANG Dong;ZHU Huibin;HU Ling;CHEN Weiwen(Piwei Institute,Science and Technology Innovation Center of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学科技创新中心广州中医药大学脾胃研究所,广东广州510405

出　　处：《中药新药与临床药理》2021年第4期533-539,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81673940);广州市科技计划项目(201607010335);广州中医药大学一流学科重点项目(广中医校办[2020]62号)。

摘　　要：目的观察白术多糖对小肠上皮细胞(IEC-6)渗透性的影响,并从其对黏附连接蛋白表达影响的角度,探讨白术多糖对肠黏膜上皮屏障的作用及机制。方法在3种实验条件下[正常含钙、含钙+多胺合成抑制剂(DFMO)、无钙培养],以检测Transwell小室酚红透过率的方法观察细胞间渗透性;以Western Blot法检测细胞黏附连接蛋白(E-cadherin、α-catenin和β-catenin)的表达。结果(1)正常含钙培养时,白术多糖(25、50、100 mg·L-1)能降低细胞间渗透性并提高黏附连接蛋白表达(P<0.05或P<0.01);(2)含钙+DFMO负荷时,细胞间渗透性增加且黏附连接蛋白表达下降(P<0.01);各剂量的白术多糖能逆转DFMO所致的细胞间渗透性增加,且对黏附连接蛋白表达下降有拮抗作用(P<0.05或P<0.01);(3)无钙培养时,细胞间渗透性增加且黏附连接蛋白表达下降(P<0.01),各剂量的白术多糖可逆转无钙培养所致的细胞间渗透性增加及E-cadherin表达降低(P<0.05或P<0.01),但对α-catenin和β-catenin表达无明显影响。结论白术多糖有增强小肠上皮屏障的作用,其机制与影响黏附连接蛋白表达有关,研究结果为探讨益气健脾中药白术对胃肠黏膜保护作用提供了参考。Objective To investigate the potential effect of Atractylodes macrocephala polysaccharides on the permeability of small intestinal epithelial cells(IEC-6),and to explore the mechanism of Atractylodes macrocephala polysaccharids on the intestinal mucosal epithelial barrier from the perspective of adhesion junction proteins expressing.Methods Under three experimental conditions(normal Ca2+-containing,Ca2+-containing+DFMO,and Ca2+-free),the transwell cell phenol red permeability was measured to observe the intercellular permeability;Western Blot was used to determine the expression of cell adhesion junction proteins(E-cadherin,α-catenin andβ-catenin).Results(1)When cultured in normal Ca2+-containing medium,the results showed that Atractylodes macrocephala polysaccharides(25,50 or 100 mg·L-1)can reduce intercellular permeability and improve the expression of adhesion junction proteins(P<0.05 or P<0.01).(2)In Ca2+-containing+DFMO culture,the intercellular permeability increased and the expression of adhesion junction proteins decreases(P<0.01).Atractylodes macrocephala polysaccharides could reverse the reduction of adhesion junction proteins expression and reduce intercellular permeability induced by polyamines synthesis inhibitor DFMO(P<0.05 or P<0.01).(3)In Ca2+-free culture,the intercellular permeability increased and the expression of adhesion junction proteins decreased(P<0.01).Atractylodes macrocephala polysaccharides can reverse the increase of intercellular permeability and decrease of E-cadherin expression caused by Ca2+-free culture(P<0.05 or P<0.01),but it had no obvious effect on the expression ofα-catenin andβ-catenin.Conclusion The results indicate that Atractylodes macrocephala polysaccharides can enhance the intestinal epithelial barrier,and the mechanism is related to its influence on the expression of adhesion junction protein.These findings provide reference for the herbs of reinforcing qi strengthen spleen for treating enteropathy characterized by mucosal damage in clinical practice.

关 键 词：白术多糖 IEC-6细胞 细胞间渗透性 上皮屏障 黏附连接蛋白 

分 类 号：R285.5[医药卫生—中药学]