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乌司他丁对肠道上皮细胞屏障损伤的保护作用分析

  

  1. 1.甘肃省人民医院 急诊科,甘肃 兰州 730000; 2.国营长风机器厂职工医院  内科,甘肃 兰州  730000
  • 出版日期:2017-11-05 发布日期:2017-11-20
  • 通讯作者: 通信作者:李永胜,Email: zf26892@126.com

Protective effect of ulinastatin on intestinal epithelial cell barrier

  1. 1.Department of Emergency,  Gansu Provincial Hospital,  Lanzhou 730000,China; 2.Department of
     Medicine, Gansu National Changfeng Machinery Factory Hospital, Lanzhou 730070, China
  • Online:2017-11-05 Published:2017-11-20
  • Contact: Corresponding author: Li Yongsheng, Email: zf26892@126.com

摘要: 目的 探讨乌司他丁(UTI)对肠道上皮细胞屏障损伤的保护作用及其机制。方法 培养Caco2细胞,并建立肿瘤坏死因子α(TNFα)诱导的单层上皮细胞模型,随机分为空白对照组、TNFα模型组、UTI低剂量组(5万U/kg)、UTI中剂量组(10万U/kg)、UTI高剂量组(20万U/kg)。采用Millicell电阻仪和多功能读板仪分别测定各组上皮细胞电阻(TER)和异硫氰酸荧光素标记的葡聚糖(FITCdextran),酶联免疫吸附测定法(ELISA)测定白细胞介素6(IL6)、IL10水平,流式细胞仪测定细胞凋亡率,免疫蛋白质印记(Western blot)技术测定紧密连接蛋白闭合蛋白(occludin)和紧密黏连蛋白1(ZO1)表达水平。结果  与空白对照组相比,TNFα模型组TER、FITCdextran、occludin和ZO1表达水平均明显下降,IL6、IL10、细胞凋亡率明显升高(P<0.05);与TNFα模型组比较,经UTI处理后,TER、FITCdextran、occludin和ZO1表达水平升高,IL6、IL10、细胞凋亡率降低(P<0.05),其中以UTI高剂量组变化最显著(P<0.05)。结论 UTI可能通过抑制炎性介质释放,调节紧密连接相关蛋白表达,抑制TNFα诱导的肠道上皮细胞屏障功能损伤。

关键词: 肠, 细胞屏障, 肿瘤坏死因子&alpha, 乌司他丁, 紧密连接蛋白

Abstract: Objective  To investigate the protective effect and mechanism of ulinastatin (UTI) on intestinal epithelial cell barrier. Methods  Caco2 cells were cultured, epithelial cells monolayer models induced by  tumor necrosis factorα(TNFα )  were established and were randomly divided into blank control group, TNFα model group, lowdose UTI group (50  000 U/kg), middledose UTI group (100  000  U/kg) and highdose UTI group (200 000  U/kg). The transepithelial electrical resistance (TER) and fluoresce  in isothiocyanate marked dextran FITCdextran in all groups were determined by Millicell electrical resistance meter and multifunctional plate reader.  Interleukin 6 (IL6) and  IL10 were determined by enzymelinked immunosorbent assay (ElISA). The apoptosis rate was measured by flow cytometry, and the expression of tight junction protein occludin and ZO1 were measured by Western blot. Results  The expression of TER, FITCdextran, occludin and ZO1 were significantly lower in TNFα model group than those in blank control group, while IL6, IL10 and apoptosis rate were significantly higher (P<0.05). After  UTI  treatment,  TER, FITCdextran, occludin and ZO1 were significantly higher than those in  TNFα model group, while IL6, IL10 and apoptosis rate were significantly lower (P<0.05).  UTI  change  in highdose group was the most significant (P<0.05). Conclusion   UTI may protect the intestinal epithelial cell barrier injury induced by TNFα through inhibiting the release of inflammatory mediators and regulating the expression of tight junction associated proteins.

Key words: intestines, cell mucosal barrier, tumor necrosis factoralpha, ulinastatin, tight junction protein