Dataset Information

Hepatocyte Rho-Associated Kinase Signaling is Required to Survive Liver Injury

ABSTRACT: Rho-associated kinases 1 and 2 (ROCK1 and ROCK2) are downstream effectors of RhoA and regulate various critical cell functions such as actomyosin contractility, apoptosis, proliferation, and cell migration. Some studies utilizing inhibitors suggested that ROCK inhibition could serve as a treatment for liver fibrosis. However, more investigation is needed to understand the role of ROCK signaling in hepatocytes with injury in vivo. Rock1fl/fl, Rock2fl/fl, and Rock1fl/fl;Rock2fl/fl mice were injected with adeno-associated virus serotype 8 (AAV8)-thyroid hormone-binding globulin (TBG)-Cre for targeted gene deletion in hepatocytes, or injected with AAV8-TBG-Null to generate littermate controls (WT). Mice were then given a 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet to induce cholestatic liver injury. Serum liver function tests, liver histology, and RNAseq analyses were performed. Mice deficient in both ROCK1 and ROCK2 (DKO) showed early mortality, decreased hepatic synthetic function, and increased hepatocellular injury compared to WT. Mice deficient in ROCK1 only (R1KO) or ROCK2 only (R2KO) demonstrated minimal differences compared to WT, indicating that the two ROCK isoforms had redundant roles in this disease model. Compared to WT, histologic analysis showed that DKO mice had significantly greater loss of liver parenchyma. RNAseq analysis indicated upregulation of apoptotic and inflammatory genes in DKO compared to WT liver. Immunostaining showed an increase in p21 and cleaved caspase 3 in DKO mice compared to WT, indicating that deletion of ROCK potentially leads to p21 overexpression and cell death. These results demonstrate that hepatocyte ROCK signaling is essential in promoting cell survival in the setting of liver cholestatic injury.

ORGANISM(S): Mus musculus

PROVIDER: GSE235163 | GEO | 2025/02/05

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Extrahepatic cholestasis leads to complex injury and repair processes that result in bile infarct formation, neutrophil infiltration, cholangiocyte and hepatocyte proliferation, extracellular matrix remodeling, and fibrosis. To identify early molecular mechanisms of injury and repair after bile duct obstruction, microarray analysis was performed on liver tissue 24 hours after bile duct ligation (BDL) or sham surgery. The most upregulated gene identified encodes plasminogen activator inhibitor 1 (PAI-1, Serpine 1), a protease inhibitor that blocks urokinase plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) activity. Because PAI-1, uPA, and tPA influence growth factor and cytokine processing as well as extracellular matrix remodeling, we evaluated the role of PAI-1 in cholestatic liver injury by comparing the injury and repair processes in wild-type (WT) and PAI-1-deficient (PAI-1-/-) mice after BDL. PAI-1-/- mice had fewer and smaller bile infarcts, less neutrophil infiltration, and higher levels of cholangiocyte and hepatocyte proliferation than WT animals after BDL. Furthermore, PAI-1-/- mice had higher levels of tPA activation and mature hepatocyte growth factor (HGF) after BDL than WT mice, suggesting that PAI-1 effects on HGF activation critically influence cholestatic liver injury. This was further supported by elevated levels of c-Met and Akt phosphorylation in PAI-1-/- mice after BDL. In conclusion, PAI-1 deficiency reduces liver injury after BDL in mice. These data suggest that inhibiting PAI-1 might attenuate liver injury in cholestatic liver diseases. Total RNA isolated using TRI Reagent (Sigma, St. Louis, MO) was purified with an RNeasy mini kit (Qiagen, Valencia, CA). Twenty micrograms cRNA was hybridized to a mouse GeneChip (U74Av2, Affymetrix, Santa Clara, CA) at the Siteman Cancer Center GeneChip facility as described by the manufacturer. Analyses used one mouse per chip. Gene expression changes were analyzed using Affymetrix MicroArray Suite 4.0 and GeneChip 3.1 Expression Analysis and Statistical Algorithms (Affymetrix). The complete methodology and full data sets for all 6 analyzed chips are available at http://bioinformatics.wustl.edu.beckerproxy.wustl.edu This study compares the injury and repair processed in wild-type mice after BDL.

Dataset Information

Hepatocyte Rho-Associated Kinase Signaling is Required to Survive Liver Injury

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets