Intestinal enteroids recapitulate the effects of short-chain fatty acids on the intestinal epithelium
Intestinal enteroids recapitulate the effects of short-chain fatty acids on the intestinal epithelium
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DOI：
10.1371/journal.pone.0230231
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发表时间：
2020-04-02
期刊：
影响因子：
3.7
通讯作者：
Breault, David T.
中科院分区：
文献类型：
--
作者：
Pearce, Sarah C.;Weber, Gregory J.;Breault, David T.
肠上皮细胞是培养的原代肠上皮细胞，其重现上皮细胞谱系发育，允许用于科学研究的更复杂和生理相关的模型。肠干细胞（ISC）的大量存在，在这些肠允许代谢物对细胞过程和产生的后代细胞的影响的研究。短链脂肪酸（SCFA）如丁酸（BUT）是胃肠道中产生的细菌代谢产物，被认为对宿主细胞有益。因此，目的是使用小鼠和人肠模型研究SCFA对肠中ISC活性、分化、屏障功能和上皮防御的生物标志物的影响。用两种浓度的乙酸盐（ACET）、丙酸盐（PROP）或BUT处理肠状体24 h。在小鼠和人模型中，相对于对照，用BUT或PROP处理的肠样组织显示通过EdU摄取的增殖减少。BUT和PROP处理后Lgr5基因表达降低，ACET处理后Lgr5基因表达增加。作为BUT和PROP治疗的结果，肠上皮细胞、潘氏细胞、杯状细胞和肠内分泌细胞的分化标志物增加。抗菌蛋白Reg3 β、Reg3 γ和Defb1的基因表达被BUT和PROP刺激，但不被ACET刺激，ACET对3D类肠组织中紧密连接基因Cldn3和Ocln的表达有更大的影响。用10 mM SCFA处理并在3D或TranswellTM模型培养物中生长的人类肠上皮细胞获得了类似的结果，尽管紧密连接受到BUT和PROP的影响，但不受单层形式的ACET的影响。此外，与ACET或对照组相比，BUT和PROP治疗24小时后增加了跨上皮电阻。总的来说，单个SCFA是细胞基因表达的有效刺激物，然而，PROP，尤其是BUT显示出驱动细胞分化和基因表达的巨大功效。
Enteroids are cultured primary intestinal epithelial cells that recapitulate epithelial lineage development allowing for a more complex and physiologically relevant model for scientific study. The large presence of intestinal stem cells (ISC) in these enteroids allows for the study of metabolite effects on cellular processes and resulting progeny cells. Short-chain fatty acids (SCFA) such as butyrate (BUT) are bacterial metabolites produced in the gastrointestinal tract that are considered to be beneficial to host cells. Therefore, the objective was to study the effects of SCFAs on biomarkers of ISC activity, differentiation, barrier function and epithelial defense in the intestine using mouse and human enteroid models. Enteroids were treated with two concentrations of acetate (ACET), propionate (PROP), or BUT for 24 h. Enteroids treated with BUT or PROP showed a decrease in proliferation via EdU uptake relative to the controls in both mouse and human models. Gene expression of Lgr5 was shown to decrease with BUT and PROP treatments, but increased with ACET. As a result of BUT and PROP treatments, there was an increase in differentiation markers for enterocyte, Paneth, goblet, and enteroendocrine cells. Gene expression of antimicrobial proteins Reg3 beta, Reg3 gamma, and Defb1 were stimulated by BUT and PROP, but not by ACET which had a greater effect on expression of tight junction genes Cldn3 and Ocln in 3D enteroids. Similar results were obtained with human enteroids treated with 10 mM SCFAs and grown in either 3D or Transwwell (TM) model cultures, although tight junctions were influenced by BUT and PROP, but not ACET in monolayer format. Furthermore, BUT and PROP treatments increased transepithelial electrical resistance after 24 h compared to ACET or control. Overall, individual SCFAs are potent stimulators of cellular gene expression, however, PROP and especially BUT show great efficacy for driving cell differentiation and gene expression.