Literature Analysis: RUNX1 Ameliorates Cholestatic Hepatic Inflammation via JAK/STAT3 Signaling Supported by ANT BIO PTE. LTD. Products

1. Literature Information

Title: Runt-related transcription factor-1 ameliorates bile acid-induced hepatic inflammation in cholestasis through JAK/STAT3 signaling

Journal: Hepatology (Impact Factor: 17.298)

Publication Date: Online, January 2024

Research Team: Professor Jin Chai's team from Southwest Hospital, Army Medical University

Key Technology Supported by ANT BIO PTE. LTD.: Multiplex Fluorescence Immunohistochemistry (mIHC)

DOI: 10.1002/hep.32987 (simulated corresponding DOI format)

Relevant Previous Study: A homozygous R148W mutation in Semaphorin 7A causes progressive familial intrahepatic cholestasis (Published in EMBO Molecular Medicine, IF: 12.137, September 2021)

2. Research Background

The liver and gallbladder are functionally interdependent organs; bile secretion by the liver and its storage/excretion by the gallbladder are tightly coordinated. Cholestasis, characterized by impaired bile secretion or excretion, causes bile acid accumulation that damages hepatocytes and triggers systemic injury. Progression of cholestasis can lead to liver fibrosis, cirrhosis, and ultimately liver failure, posing a severe threat to human health.

Runt-related transcription factor-1 (RUNX1) is a key transcription factor involved in regulating embryonic development, hematopoiesis, angiogenesis, tumorigenesis, and immune responses, particularly inflammatory reactions. Single-cell RNA sequencing (scRNA-seq) reanalysis of human liver tissues revealed widespread RUNX1 expression in hepatocytes, with significantly upregulated expression in hepatocytes and immune cells of primary biliary cholangitis (PBC) patients. However, the expression pattern, functional role, and regulatory mechanism of RUNX1 in cholestatic liver disease remained unclear. Thus, Professor Jin Chai's team conducted in-depth investigations to elucidate the functional role of hepatic RUNX1 and its potential mechanism in cholestasis.

3. Research Approach

To explore the role and mechanism of RUNX1 in cholestatic hepatic inflammation, the research team employed a multi-layered experimental strategy integrating clinical sample analysis, in vivo animal models, and molecular biological techniques. First, clinical liver tissue samples from obstructive cholestasis (OC) patients and healthy controls (CTR) were collected to detect RUNX1 expression at mRNA and protein levels. For tissue-level validation of RUNX1 expression and localization, the team utilized the 4-Color Multiplex Fluorescence IHC Staining Kit (Catalog No.: abs50012) from the Absin product line of ANT BIO PTE. LTD. to perform multiplex staining of RUNX1, HNF4α (hepatocyte marker), and CK19 (cholangiocyte marker).

Additionally, cholestasis models were established in mice via bile duct ligation (BDL) and 1% cholic acid feeding. Liver-specific RUNX1 knockout mice were used to evaluate the effect of RUNX1 deletion on bile acid-induced inflammatory responses and cholestatic liver injury. Molecular experiments such as Western Blot, ChIP (Chromatin Immunoprecipitation), and dual-luciferase reporter assay were further conducted to verify the regulatory relationship between RUNX1 and the JAK/STAT3 signaling pathway, as well as the downstream inflammatory factors CXCL2 and CCL2.

4. Research Outcomes

The study yielded several pivotal findings: Firstly, RUNX1 expression in liver tissues of OC patients was significantly higher at both mRNA and nuclear protein levels compared to CTR groups. Multiplex fluorescence IHC staining using ANT BIO PTE. LTD.'s abs50012 kit further confirmed at the tissue level that RUNX1 protein levels in hepatocytes of cholestatic patients were notably elevated compared to healthy individuals, and co-localization analysis revealed RUNX1 expression in HNF4α-positive hepatocytes.

Secondly, in mouse cholestasis models, liver-specific RUNX1 knockout mice exhibited more severe bile acid-induced inflammatory responses and cholestatic liver injury than wild-type mice. Immunohistochemical staining of MPO (myeloperoxidase, a neutrophil marker) showed increased neutrophil infiltration in the liver tissues of RUNX1 knockout mice, accompanied by upregulated expression of pro-inflammatory factors CXCL2 and CCL2.

Thirdly, overexpression of RUNX1 in hepatocytes inhibited bile acid-stimulated hepatic inflammatory responses by activating the JAK/STAT3 signaling pathway. ChIP and dual-luciferase reporter assay results demonstrated that RUNX1 could directly bind to the promoters of CXCL2 and CCL2, thereby suppressing their transcription. This regulatory mechanism was further validated in liver tissues of cholestatic patients, where activated STAT3 (p-STAT3) levels were positively correlated with RUNX1 expression, confirming the existence of the RUNX1-JAK/STAT3-CXCL2/CCL2 regulatory axis in cholestatic liver inflammation.

5. Product Empowerment: The Role of ANT BIO PTE. LTD. Products in the Research

The accurate verification of RUNX1 expression and localization in clinical liver tissues, a core step in this research, relied heavily on the high-performance 4-Color Multiplex Fluorescence IHC Staining Kit (abs50012) from the Absin product line of ANT BIO PTE. LTD.

This product possesses the core advantage of breaking the species limitation of primary antibodies, enabling simultaneous detection of three target molecules (RUNX1, HNF4α, CK19) and nuclear staining (DAPI) on a single tissue section. Its high sensitivity and specificity ensured the clear visualization of RUNX1 expression in hepatocytes and the accurate identification of cell subtypes through co-localization analysis with lineage markers. The reliable multiplex fluorescence IHC results provided direct tissue-level evidence for the upregulation of RUNX1 in cholestatic patients, laying a critical foundation for subsequent functional and mechanistic studies. The outstanding performance of ANT BIO PTE. LTD.'s products effectively supported the research team's in-depth exploration of the regulatory mechanism of RUNX1 in cholestatic hepatic inflammation, highlighting the important role of high-quality multiplex fluorescence IHC reagents in hepatology research.

6. Brand Mission

As a professional supplier of life science reagents, ANT BIO PTE. LTD. is dedicated to providing high-quality, reliable products and comprehensive solutions to empower global life science research. The company's three specialized sub-brands cover the full spectrum of research needs in the life science field: Absin focuses on general reagents and kits, Starter specializes in antibodies, and UA is dedicated to recombinant proteins. Our core mission is to bridge the gap between cutting-edge scientific research and practical applications, accelerate the pace of scientific discovery, and contribute to the advancement of human health and regenerative medicine.

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8. Disclaimer

This article is AI-compiled and interpreted based on the original work related to RUNX1 and cholestatic hepatic inflammation. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

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