KLF4 通过调节 NLRP3 信号通路与 TXNIP 相互作用，调节溃疡性结肠炎中的细胞焦亡。

KLF4 interacts with TXNIP to modulate the pyroptosis in ulcerative colitis via regulating NLRP3 signaling.

机构信息

Department of Pediatrics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People's Republic of China.

Shandong Provincial Clinical Research Center for Children's Health and Disease Office, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People's Republic of China.

出版信息

Immun Inflamm Dis. 2024 Feb;12(2):e1199. doi: 10.1002/iid3.1199.

DOI:10.1002/iid3.1199

PMID:38411328

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10898204/

Abstract

INTRODUCTION

Ulcerative colitis (UC) is one of the most common diseases in the gastrointestinal tract related to abnormal inflammation. Pyroptosis, which is characterized by the formation of inflammasome, activation of caspase-1, and separation of N- and C-terminus of gasdermin D (GSDMD), and may be involved in the pathogenesis of IBD. Krüppel-like factor 4 (KLF4) is a zinc finger transcription factor expressed in differentiated epithelial cells. KLF4 mediates proinflammatory signaling in macrophages. Here, we tested whether KLF4 is functional in pyroptosis of UC.

METHODS

In human UC tissues and/or lipopolysaccharide (LPS)/adenosine 5-triphosphate (ATP) stimulation human colon epithelial cells, KLF4, TXNIP, Cleave-Caspase-1, and GSDMD expression were detected through quantitative reverse transcription polymerase chain reaction (PCR), immunohistochemical and western blot assay. Interleukin (IL)-1β and IL-18 levels were quantified by enzyme-linked immunosorbent assay. We successfully constructed a KLF4-silenced colon epithelial cell line using an adenovirus vector. We apply the UCSC and JASPAR to predict the KLF4 binding sites in the promoter region of TXNIP.

RESULTS

In human UC tissues and/or LPS/ATP stimulation human colon epithelial cells, KLF4, TXNIP, Caspase-1, and GSDMD expression level were significantly elevated via quantitative reverse transcription PCR, immunohistochemical and western blot assay. Moreover, We identified that there is an interaction between KLF4 and TXNIP through Yeast double hybrid assay and CO-IP assay. We successfully constructed a KLF4-silenced human intestinal epithelial cell line. In LPS/ATP stimulation KLF4-silenced human intestinal epithelial cells, KLF4, TXNIP, Cleave Caspase-1, ASC, and GSDMD expression level were significantly decreased via quantitative reverse transcription PCR.

CONCLUSION

Our results confirm that KLF4 can positively regulate the expression of TXNIP and regulate the pyroptosis process of UC through the TXNIP/NLRP3 pathway.

摘要

简介

溃疡性结肠炎（UC）是胃肠道最常见的与异常炎症相关的疾病之一。细胞焦亡，其特征在于炎性小体的形成、半胱天冬酶-1 的激活以及 Gasdermin D（GSDMD）的 N-和 C-末端的分离，可能参与了 IBD 的发病机制。Krüppel 样因子 4（KLF4）是一种在分化上皮细胞中表达的锌指转录因子。KLF4 介导巨噬细胞中的促炎信号。在这里，我们测试了 KLF4 是否在 UC 的细胞焦亡中起作用。

方法

在人类 UC 组织和/或脂多糖（LPS）/三磷酸腺苷（ATP）刺激的人结肠上皮细胞中，通过定量逆转录聚合酶链反应（PCR）、免疫组织化学和 Western blot 检测 KLF4、TXNIP、Cleaved-Caspase-1 和 GSDMD 的表达。通过酶联免疫吸附试验定量测定白细胞介素（IL）-1β和 IL-18 的水平。我们成功地使用腺病毒载体构建了 KLF4 沉默的结肠上皮细胞系。我们应用 UCSC 和 JASPAR 预测 TXNIP 启动子区域中 KLF4 的结合位点。

结果

在人类 UC 组织和/或 LPS/ATP 刺激的人结肠上皮细胞中，通过定量逆转录 PCR、免疫组织化学和 Western blot 检测，KLF4、TXNIP、Caspase-1 和 GSDMD 的表达水平显著升高。此外，我们通过酵母双杂交实验和 CO-IP 实验证实了 KLF4 和 TXNIP 之间存在相互作用。我们成功构建了 KLF4 沉默的人肠道上皮细胞系。在 LPS/ATP 刺激的 KLF4 沉默的人肠道上皮细胞中，通过定量逆转录 PCR，KLF4、TXNIP、Cleaved Caspase-1、ASC 和 GSDMD 的表达水平显著降低。