THU-058-Gene profiling of toll-like recepter signaling pathways in patients with acute-on-chronic liver failure

Abstract BackgroundAcute-on-chronic liver failure (ACLF) is a severe complication of cirrhosis, which seriously endanger human life and health. Yiqijianpi decoction (YQJPF) is a Traditional Chinese Medicine (TCM) formula that has been widely used in the treatment of liver failure with significant effect. The purpose of this study was to investigate the active components and mechanism to ameliorate acute on chronic liver failure(ACLF).MethodsLPS combined with D-Gal was used to establish the rat model of ACLF, pathological examination was detected by H&E staining, liver function test was assayed by ELISA. LPS was used to induce hepatocytes injury in vitro. Cell proliferation assay, TUNEL assay were used in human hepatic L02 cells. The active components and putative targets of YQJPF were predicted by network pharmacology approach and GEO analysis. Functional and pathway enrichment analysis were presented by using String, Cytoscape and Metascape. Further, experimental validation was done to verify the effect of YQJPF on PI3K/AKT-HIF-1ɑ and apoptosis-related signaling pathways by using Immunohistochemistry and Western blotting.ResultsAfter being treated with YQJPF, the rat liver injury and fibrosis were alleviated, and Cell Counting Kit-8 assay, TUNEL assay indicated YQJPF also inhibited the apoptosis in hepatic L02 cells. Through network pharmacologic analysis, 135 active components in YQJPF decoction, 573 known therapeutic targets and 2940 liver failure-related human genes were identified. 163 gene symbols maybe the key for liver failure treatment by YQJPF decoction. VEGF-A was hub gene in PPI network. The KEGG pathway and GO enrichment analyses indicated that the PI3K/AKT, HIF-1 signaling pathways were the prominently enriched signaling pathways. In vivo, YQJPF upregulated the expression of PI3K/AKT-HIF1-ɑ and VEGF-A. Moreover apoptosis pathway were verified by up-regulating Bcl2 expression and down-regulating Bax expression in vivo and in vitro.ConclusionYQJPF is beneficial for alleviating liver failure, may regulate hypoxic liver injury through PI3K/AKT-HIF1α dependent apoptosis pathway.

Download Full-text

Gene Profiling of Toll-like Receptor Signaling Pathways in Neutrophils of Patients With Acute-on-chronic Liver Failure

10.21203/rs.3.rs-541965/v1 ◽

2021 ◽

Author(s):

Yi Zhang ◽

Wei Wu ◽

Yijie Wang ◽

Lingjia Tong ◽

Meng Hong ◽

...

Keyword(s):

Liver Failure ◽

Signaling Pathways ◽

Signaling Pathway ◽

Tissue Injury ◽

Fold Change ◽

Chronic Liver ◽

Chronic Liver Failure ◽

Tlr Signaling ◽

Compensated Cirrhosis ◽

Tlr Signaling Pathway

Abstract Objectives: Toll-like receptors (TLRs) of neutrophils play a crucial role in detecting pathogens and organ/tissue injury in acute-on-chronic liver failure (ACLF). However, little is known about the exact mechanisms and the potential signaling pathways. The aim of this study was to investigate alterations of TLR signaling pathways in neutrophils of ACLF patients.Methods: Twenty-seven patients with compensated cirrhosis (n=9), de-compensated cirrhosis (n=9) and ACLF (n=9) were enrolled in the study. Neutrophils were isolated, and alterations in TLR signaling pathways were evaluated using an RT² Profiler™ PCR Array. The fold change for each gene (2(-∆∆CT)) was compared among the groups. Genes with a fold change ratio of ≥2 or ≤0.5 along with a p value of < 0.05 were considered to be differentially expressed.Results: A total of 17 genes were up-regulated in neutrophils from patients with compensated cirrhosis, which were mainly distributed in adaptors, TLR-interacting proteins and downstream pathways. Six genes were detected in patients with de-compensated cirrhosis. A trend of down-regulation of genes in the TLR signaling pathway was observed in neutrophils of patients with cirrhosis and ACLF. TLR3, IFNG, IL1B, TBK1, CCL2 and LTA were downregulated in neutrophils. Moreover, CD14 and IL10 were up regulated in neutrophils of ACLF patients.Conclusions: TLR signaling pathway genes were differentially regulated in neutrophils between cirrhosis and ACLF. In ACLF patients, there was defective expression of TLR3 and IFN, along with enhanced CD14 and IL10 expression, characterized by transcriptional alterations of neutrophils.

Download Full-text