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应激因素对肠上皮损伤和再生的影响。

Influence of stress factors on intestinal epithelial injury and regeneration.

作者信息

Lee Carol, Minich Adam, Li Bo, Miyake Hiromu, Seo Shogo, Pierro Agostino

机构信息

Division of General and Thoracic Surgery, Program of Translational Medicine, The Hospital for Sick Children, 1526-555 University Ave, Toronto, ON M5G 1X8, Canada.

出版信息

Pediatr Surg Int. 2018 Feb;34(2):155-160. doi: 10.1007/s00383-017-4183-3. Epub 2017 Oct 10.

Abstract

PURPOSE

Lgr5+ intestinal epithelial stem cells (ISCs) crucial for intestinal epithelial regeneration are impaired during necrotizing enterocolitis. This study aims to investigate the influence of different stressors on intestinal epithelial injury and regeneration in vitro.

METHODS

Intestinal epithelial cells (IEC-18) were exposed to stressors such as lipopolysaccharide, hydrogen peroxide, and serum. Cell viability was assessed using MTT assay at 18 and 24 h. IL-6 and Lgr5 gene expressions were measured using qPCR.

RESULTS

IEC-18 cell viability decreased 18 h following administration of lipopolysaccharide, hydrogen peroxide, and low serum concentration. However, after 24 h, the decrease in cell viability was observed only in higher, but not in lower concentrations of lipopolysaccharide and hydrogen peroxide. IL-6 expression increased in all groups compared to control. Lgr5 expression was up-regulated in cells exposed to a single stressor, but down-regulated when multiple stressors were administered.

CONCLUSION

Lipopolysaccharide, hydrogen peroxide, or low serum induced IEC-18 injury. The upregulation of Lgr5 expression after exposure to a single stressor suggests that minor injury to IEC-18 induces Lgr5+ ISCs to stimulate repair. Conversely, when IEC-18 cells were exposed to multiple stressors, Lgr5 expression was reduced. We speculate that this finding is similar to what happens in NEC when multiple stressors cause impairment of intestinal epithelium regeneration.

摘要

目的

对肠道上皮再生至关重要的Lgr5⁺肠道上皮干细胞(ISCs)在坏死性小肠结肠炎期间会受到损害。本研究旨在探讨不同应激源对体外肠道上皮损伤和再生的影响。

方法

将肠道上皮细胞(IEC-18)暴露于脂多糖、过氧化氢和血清等应激源中。在18小时和24小时时使用MTT法评估细胞活力。使用qPCR测量IL-6和Lgr5基因表达。

结果

给予脂多糖、过氧化氢和低血清浓度后18小时,IEC-18细胞活力下降。然而,24小时后,仅在较高浓度而非较低浓度的脂多糖和过氧化氢中观察到细胞活力下降。与对照组相比,所有组的IL-6表达均增加。暴露于单一应激源的细胞中Lgr5表达上调,但给予多种应激源时则下调。

结论

脂多糖、过氧化氢或低血清诱导IEC-18损伤。暴露于单一应激源后Lgr5表达上调表明IEC-18的轻微损伤会诱导Lgr5⁺ ISCs刺激修复。相反,当IEC-18细胞暴露于多种应激源时,Lgr5表达降低。我们推测这一发现与坏死性小肠结肠炎中多种应激源导致肠道上皮再生受损的情况相似。

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