摘要
目的探究高盐饮食对肠屏障的影响及细胞自噬变化。方法C57BL/6雄性小鼠随机分为正常饮食(NSD)组(n=9),高盐饮食(HSD)组(n=9),喂养12周采用FITC-dextran法测定肠屏障通透性,取结肠组织观察HE病理染色,Western blot法测定紧密连接蛋白ZO-1、Occludin,自噬相关蛋白ATG5,LC3Ⅱ/Ⅰ的蛋白表达水平。分别以0、50、80、100mmol/L浓度的Na Cl处理Coco2结肠癌细胞,Western blotting检测细胞自噬相关蛋白ATG5(autophagy-related gene5)、LC3Ⅱ/Ⅰ(microtubule-associated protein light chain 3Ⅱ/Ⅰ)表达。敲低Coco2细胞ATG5表达,观察紧密连接蛋白ZO-1、Occludin表达。结果与NSD组相比,HSD组血浆FITC-dextran浓度显著增加(P<0.05),结肠绒毛缺损改变,杯状细胞减少,紧密连接蛋白Z0-1、Occludin和自噬蛋白ATG5、LC3Ⅱ/Ⅰ表达水平明显降低(P<0.05)。随着Na Cl浓度增加,ATG5、LC3Ⅱ/Ⅰ蛋白在Coco2细胞中的表达逐渐降低,敲低ATG5表达后,ZO-1、Occludin蛋白表达明显降低(P<0.05)。结论高盐饮食可能通过下调ATG5表达抑制细胞自噬进而损伤肠屏障功能。
Objective To explore the effects of high-salt diet on intestinal barrier and cell autophagy.Methods Male C57 BL/6 mice were randomly divided into the normal diet(NSD)group(n=9)and the high-salt diet(HSD)group(n=9).After 12 weeks of feeding,FITC-dextran method was used to measure intestinal barrier permeability,colon tissues were taken for observation of HE pathological staining,and tight junction protein ZO-1,Occludin,autophag-related proteins Atg5 and LC3Ⅱ/Ⅰwere determined by western blot.Coco2 colon cancer cells were treated with Na Cl at concentrations of 0,50,80 and 100 mmol/L,and the expression of autophagy-related proteins Atg5 and LC3Ⅱ/Ⅰwere detected by Western blotting.We observed the expression of ATG5 in Coco2 cells was decreased,as well as the expression of tight junction protein zo-1 and Occludin.Results Compared with NSD group,plasma FITC-dextran concentration in the HSD group was significantly increased(P<0.05),the colonic villi was defective,and goblet cells decreased,meanwhile the expression levels of tight junction protein Z0-1,Occludin and autophag-related proteins ATG5 and LC3Ⅱ/Ⅰwere significantly decreased(P<0.05).Along with the increase of Na Cl concentration,the expression of Atg5 and LC3Ⅱ/Ⅰproteins in Coco2 cells gradually decreased,and the expression of ZO-1 and Occludin proteins significantly decreased after Atg5 expression was reduced in Coco2 cell(P<0.05).Conclusion High-salt diet inhibits autophagy by down-regulating the expression of ATG5 and thus damages the intestinal barrier function.
作者
雷超
刘聪
李晓媚
李晨
刘志华
刘亭
LEI Chao;LIU Cong;LI Xiaomei;LI Chen;LIU Zhihua;LIU Ting(Department of Center Laboratory,the Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China)
出处
《现代医院》
2021年第1期150-153,共4页
Modern Hospitals
基金
国家自然科学基金(816704800)
广东省自然科学基金资助项目(2018A0303101680)。
关键词
肠屏障功能
肠紧密连接
高盐饮食
细胞自噬
ATG5
Intestinal Barrier Function
Intestinal Tight Junction
High-salt Diet
Autophagy
ATG5
