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尿毒症血清对人结肠细胞屏障功能和炎症的体外作用。

In vitro effect of uremic serum on barrier function and inflammation in human colonocytes.

作者信息

Andrade Laila Santos de, Dalboni Maria Aparecida, Carvalho José Tarcisio Giffoni de, Grabulosa Caren Cristina, Pereira Natalia Barros Ferreira, Aoike Danilo Takashi, Cuppari Lilian

机构信息

Universidade Federal de São Paulo, Programa de Pós-Graduação em Nutrição, São Paulo, SP, Brasil.

Universidade Federal de São Paulo, Departamento de medicina, Divisão de Nefrologia, São Paulo, SP, Brasil.

出版信息

J Bras Nefrol. 2018 Jul-Sep;40(3):217-224. doi: 10.1590/2175-8239-jbn-3949. Epub 2018 Jun 18.

Abstract

INTRODUCTION

In chronic kidney disease (CKD), it has been suggested that alterations within the gut are associated with an inflammatory state and uremic toxicity. Studies suggest that uremia may impair the function of the intestinal barrier via the promotion of increased intestinal permeability. To understand the mechanisms that are involved in intestinal barrier damage in the setting of uremia, we evaluated the in vitro effect of uremic serum on transepithelial electrical resistance (TER), inflammation, and apoptosis in intestinal epithelial cells (T84).

METHODS

Pools of serum from healthy individuals, patients not on dialysis, and patients on hemodialysis (Pre-HD and Post-HD) were prepared. T84 cells were incubated for 24 h in medium, of which 10% consisted of the pooled serum from each group. After incubation, the TER was measured and the following parameters were determined by flow cytometry: expression of toll-like receptors (TLRs), production of reactive oxygen species (ROS), and apoptosis. The level of IL-6 in the culture supernatant was determined by ELISA.

RESULTS

No difference was observed among the groups with respect to TER, apoptosis, and ROS or the expression of TLR-2, TLR-4, and TLR-9. IL-6 secretion was higher (p < 0.001) in cells that were incubated with pre- and post-HD serum.

CONCLUSION

The results that were obtained from this model suggest that uremic serum per se does not seem to impair the integrity of intestinal epithelial cells. The increased IL-6 secretion by cells that were incubated with HD serum suggests a potential effect of uremia in the intestinal inflammatory response.

摘要

引言

在慢性肾脏病(CKD)中,已有研究表明肠道内的改变与炎症状态和尿毒症毒性相关。研究提示,尿毒症可能通过促进肠通透性增加而损害肠屏障功能。为了解尿毒症状态下肠屏障损伤的相关机制,我们评估了尿毒症血清对肠上皮细胞(T84)跨上皮电阻(TER)、炎症及凋亡的体外作用。

方法

制备健康个体、未透析患者及血液透析患者(透析前和透析后)的混合血清。将T84细胞在含10%各组混合血清的培养基中孵育24小时。孵育后,测量TER,并通过流式细胞术测定以下参数:Toll样受体(TLR)的表达、活性氧(ROS)的产生及凋亡。采用酶联免疫吸附测定法(ELISA)测定培养上清液中白细胞介素-6(IL-6)的水平。

结果

各组在TER、凋亡、ROS或TLR-2、TLR-4及TLR-9的表达方面未观察到差异。与透析前和透析后血清孵育的细胞中IL-6分泌更高(p<0.001)。

结论

该模型获得的结果提示,尿毒症血清本身似乎并未损害肠上皮细胞的完整性。与透析血清孵育的细胞中IL-6分泌增加提示尿毒症在肠道炎症反应中可能发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d236/6533953/3e0e33e83eaa/2175-8239-jbn-3949-gf01.jpg

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