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1 μg/mL LPS，给药组分别加入10%含药血清、10 μmol/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-1β mRNA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-1β mRNA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, 1 μg/mL LPS was added to the other groups, 10% drug-containing serum and 10 μmol/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-1β were 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-1β were 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-1β in 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.

R285.5

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）