亮菌多糖对LPS诱导的肠上皮细胞炎性损伤的保护作用及机制探讨  

Protective effect and mechanism of Armillariella polysaccharide on LPS induced inflammatory injury of intestinal epithelial cells

作　　者：张梅[1,2] 吴悦[1,2] 苏丽[1,2] 朱耀东 李平 王婷[1,2] Zhang Mei;Wu Yue;Su Li(Dept of Integrative Medicine Oncology of First Affiliated Hospital of Anhui Medical University,Hefei 230022;The Medicine(TCM)-Integrated Cancer Center of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学中西医结合肿瘤中心,合肥230022 [2]安徽医科大学第一附属医院中西医结合肿瘤科,合肥230022

出　　处：《安徽医科大学学报》2021年第4期533-538,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81774051)。

摘　　要：目的观察在脂多糖(LPS)诱导的IEC-6肠上皮细胞损伤的情况下,亮菌多糖(ATPS)对细胞活性、闭合素(Occludin)、细胞紧密连接蛋白1(ZO-1)的影响,并探讨ATPS对肠黏膜屏障损伤的保护机制。方法利用LPS构建的IEC-6肠上皮细胞体外损伤模型,将细胞随机分为对照组、LPS组、LPS+ATPS组、LPS+Matrine组(阳性对照),使用普通显微镜和MTT法观察细胞的活性;免疫荧光和流式细胞术检测细胞的凋亡情况;Western blot检测膜蛋白ZO-1和跨膜蛋白Occludin的表达水平。同时Western blot检测与介导细胞凋亡密切相关的内质网应激(ERS)通路上磷酸化蛋白激酶R样内质网调节激酶(p-PERK)、磷酸化真核细胞翻译起始因子(p-eIF2α)、葡萄糖调节蛋白78(GRP78)、内质网应激相关蛋白(CHOP)以及调节肠黏膜损伤重要分子β-抑制蛋白1(Arrb1)的表达水平。结果与对照组、LPS组比较,随着ATPS的浓度增加,LPS+ATPS组中细胞的活性越多且免疫荧光和流式检测也表明细胞的凋亡率减少;Occludin、ZO-1的表达水平增多,明显高于LPS组;与LPS组相比,LPS+ATPS组中p-PERK、p-eIF2α、GRP78、CHOP及Arrb1蛋白的水平降低。结论ATPS可通过抑制Occludin、ZO-1的破坏来保护LPS介导的肠上皮细胞炎性损伤,其机制可能与介导细胞凋亡的内质网应激蛋白以及调节肠黏膜损伤的重要分子Arrb1有关。Objective To investigate the effects of Armillariella tabescens polysaccharides(ATPS)on cell activity,occludin and ZO-1 in IEC-6 intestinal epithelial cells induced by lipopolysaccharide(LPS),and to reveal the protective mechanism of Armillariella tabescens polysaccharides on intestinal mucosal barrier injury.Methods IEC-6 intestinal epithelial cell injury model induced by LPS was established.The injured cells were randomly divided into control group,LPS group,LPS+ATPS group and LPS+matrine group.The cell viability was observed by microscope and MTT;the cell apoptosis was detected by immunofluorescence and flow cytometry;the expression of ZO-1 and occludin was detected by Western blot.Meanwhile,Western blot was used to detect the expression of p-PERK,p-eIF2α,GRP78,CHOP and Arrb1 proteins.Results With the increase of ATPS concentration,the cell activity in the LPS+ATPS group increased,and the apoptosis rate decreased by immunofluorescence and flow cytometry compared with control and LPS group.The expression levels of occludin and ZO-1 protein were significantly higher in LPS+ATPS group than those in LPS group.The levels of p-PERK,p-eIF2α,GRP78,chop and arrb1 protein in LPS+ATPS group decreased compared with LPS group.Conclusion Armillariella polysaccharide can protect intestinal epithelial cells from LPS mediated inflammatory injury by inhibiting the damage of occludin and ZO-1 proteins.The mechanism may be related to endoplasmic reticulum stress protein which mediates apoptosis and Arrb1(an important molecule)regulating intestinal mucosal injury.

关 键 词：IEC-6肠上皮细胞损伤 脂多糖 亮菌多糖 紧密连接 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]