扶肾颗粒改善肠上皮细胞屏障损伤的作用机制研究  

Mechanism of Fushen Granule on improving intestinal epithelial barrier injury

作　　者：姜晨[1] 杨洪涛[1] 耿芳 刘睿昕 忽星歌 JIANG Chen;YANG Hong-tao;GENG Fang;LIU Rui-xin;HU Xing-ge(Department of Nephrology,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;Tianjin university of Traditional Chinese Medicine,Tianjin 301608,China)

机构地区：[1]天津中医药大学第一附属医院肾病科,天津300381 [2]天津中医药大学,天津301608

出　　处：《中草药》2020年第21期5496-5500,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81673909);国家自然科学基金青年项目(81302920)。

摘　　要：目的探讨扶肾颗粒对脂多糖(LPS)诱导的肠上皮(IEC-6)细胞屏障损伤的保护机制。方法IEC-6细胞分为对照组、模型组、扶肾颗粒组、p38MAPK抑制剂组,除对照组外,其余各组加入1μg/mL LPS,给药组分别加入10%含药血清、10μmol/L p38MAPK抑制剂(SB203580)。流式细胞仪检测细胞凋亡情况;Transwell法检测细胞屏障通透性、跨膜电阻;Real-time PCR法检测细胞ICAM-1、IL-1βmRNA水平;免疫印迹检测胞质内细胞紧密连接蛋白Occludin、ZO-1和磷酸化p38MAPK、DUSP1蛋白表达的变化。结果与对照组比较,模型组IEC-6细胞凋亡、炎症因子ICAM-1和IL-1β水平显著增加(P<0.01),紧密连接蛋白Occludin、ZO-1表达显著降低(P<0.01),肠黏膜通透性显著增加(P<0.05)。与模型组比较,扶肾颗粒能明显抑制IEC-6细胞内炎症因子ICAM-1、IL-1βmRNA水平,降低肠黏膜屏障通透性(P<0.05),上调DUSP1、Occludin、ZO-1水平(P<0.05、0.01),抑制p38MAPK表达(P<0.01),与p38MAPK抑制剂作用一致。结论扶肾颗粒可能通过调控DUSP1,抑制p38MAPK信号通路的激活,上调Occludin和ZO-1,改善IEC-6细胞紧密连接,从而改善LPS引起的细胞屏障功能破坏。Objective To investigate the protection mechanism of Fushen Granule on LPS-induced intestinal epithelial cell(IEC-6)barrier injury.Methods IEC-6 were divided into control group,model group,Fushen granule group,and p38MAPK inhibitor group.In addition to the control group,1μg/mL LPS was added to the other groups,10%drug-containing serum and 10μmol/L SB203580 were respectively added to the administration group.The apoptosis of IEC-6 were detected by flow cytometry;The permeability and transmembrane resistance of IEC-6 barrier were detected by Transwell assay;The mRNA levels of ICAM-1 and IL-1βwere assessed by Real-time PCR;The expressions of occludin,ZO-1,p38MAPK,and DUSP1 were detected by western blotting.Results Compared with the control group,the apoptosis of IEC-6 were significantly increased in model group(P<0.01),the levels of ICAM-1 and IL-1βwere significantly increased(P<0.01),the occludin and ZO-1 were significantly decreased(P<0.01),the intestinal permeability was significantly increased(P<0.05),the mRNA levels of ICAM-1 and IL-1βin IEC-6 were significantly inhibited,the permeability of intestinal mucosal barrier was reduced(P<0.05),the expressions of DUSP1,occludin and ZO-1were increased(P<0.05,0.01),the expression of p38MAPK was inhibited(P<0.01),which showed a similar effect with p38MAPK inhibitor.Conclusion Fushen Granule could regulate DUSP1 which inhibits the activation of p38MAPK signaling pathway,up-regulate Occludin and ZO-1,improve the tight junction of IEC-6,thereby improving the cell barrier function damage caused by LPS.

关 键 词：扶肾颗粒 大鼠肠上皮细胞 肠黏膜屏障 紧密连接 P38MAPK 

分 类 号：R285.5[医药卫生—中药学]