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Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation.

Lan T, Kisseleva T, Brenner DA - PLoS ONE (2015)

Bottom Line: Furthermore, the mRNAs of proliferative and pro-fibrotic genes were downregulated in NOX1 and NOX4 knock-out activated HSCs (cultured on plastic for 5 days).Finally, NOX1 and NOX4 protein levels were increased in human livers with cirrhosis compared with normal controls.Thus, NOX1 and NOX4 signaling mediates the pathogenesis of liver fibrosis, including the direct activation of HSC.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of California San Diego, La Jolla, California, United States of America; Vascular Biology Research Institute, Guangdong Pharmaceutical University, Guangzhou, China.

ABSTRACT
Reactive oxygen species (ROS) produced by nicotinamide adenine dinucleotide phosphate oxidase (NOX) play a key role in liver injury and fibrosis. Previous studies demonstrated that GKT137831, a dual NOX1/4 inhibitor, attenuated liver fibrosis in mice as well as pro-fibrotic genes in hepatic stellate cells (HSCs) as well as hepatocyte apoptosis. The effect of NOX1 and NOX4 deficiency in liver fibrosis is unclear, and has never been directly compared. HSCs are the primary myofibroblasts in the pathogenesis of liver fibrosis. Therefore, we aimed to determine the role of NOX1 and NOX4 in liver fibrosis, and investigated whether NOX1 and NOX4 signaling mediates liver fibrosis by regulating HSC activation. Mice were treated with carbon tetrachloride (CCl4) to induce liver fibrosis. Deficiency of either NOX1 or NOX4 attenuates liver injury, inflammation, and fibrosis after CCl4 compared to wild-type mice. NOX1 or NOX4 deficiency reduced lipid peroxidation and ROS production in mice with liver fibrosis. NOX1 and NOX4 deficiency are approximately equally effective in preventing liver injury in the mice. The NOX1/4 dual inhibitor GKT137831 suppressed ROS production as well as inflammatory and proliferative genes induced by lipopolysaccharide (LPS), platelet-derived growth factor (PDGF), or sonic hedgehog (Shh) in primary mouse HSCs. Furthermore, the mRNAs of proliferative and pro-fibrotic genes were downregulated in NOX1 and NOX4 knock-out activated HSCs (cultured on plastic for 5 days). Finally, NOX1 and NOX4 protein levels were increased in human livers with cirrhosis compared with normal controls. Thus, NOX1 and NOX4 signaling mediates the pathogenesis of liver fibrosis, including the direct activation of HSC.

No MeSH data available.


Related in: MedlinePlus

Both NOX1 and NOX4 mediate proliferative and fibrogenic responses in HSCs.HSCs from WT, NOX1KO and NOX4KO mice were cultured for 1 day (quiescent HSCs) and 5 days (activated HSCs) with 10% FBS in DMEM. mRNA expression of proliferative genes (A) and fibrogenic genes (B) were measured by quantitative real-time PCR. HPRT was used as an internal control. *P <0.05, **P < 0.01, ***P < 0.001 vs activated WT HSCs control.
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pone.0129743.g007: Both NOX1 and NOX4 mediate proliferative and fibrogenic responses in HSCs.HSCs from WT, NOX1KO and NOX4KO mice were cultured for 1 day (quiescent HSCs) and 5 days (activated HSCs) with 10% FBS in DMEM. mRNA expression of proliferative genes (A) and fibrogenic genes (B) were measured by quantitative real-time PCR. HPRT was used as an internal control. *P <0.05, **P < 0.01, ***P < 0.001 vs activated WT HSCs control.

Mentions: We next investigated whether similar findings are observed in cultured HSCs treated with PDGF. As expected, the number of Ki67+HSCs isolated from WT was increased by PDGF treatment. However, PDGF produced significantly fewer Ki67+HSCs from NOX1KO and NOX4KO mice (Fig 6B and 6D). Furthermore, upregulation of proliferative genes (Cyclin D1, Bcl-2, PCNA and PDGFRB) was observed in activated WT HSCs (5 day plastic culture with 10% FBS medium). Proliferation of HSCs was attenuated by NOX1 and NOX4 deficiency. However, there was no change of proliferative genes in quiescent HSCs (1 day plastic culture) (Fig 7A). These results suggest that NOX1 and NOX4 mediate proliferation of HSCs in response to proliferative stimulation.


Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation.

Lan T, Kisseleva T, Brenner DA - PLoS ONE (2015)

Both NOX1 and NOX4 mediate proliferative and fibrogenic responses in HSCs.HSCs from WT, NOX1KO and NOX4KO mice were cultured for 1 day (quiescent HSCs) and 5 days (activated HSCs) with 10% FBS in DMEM. mRNA expression of proliferative genes (A) and fibrogenic genes (B) were measured by quantitative real-time PCR. HPRT was used as an internal control. *P <0.05, **P < 0.01, ***P < 0.001 vs activated WT HSCs control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519306&req=5

pone.0129743.g007: Both NOX1 and NOX4 mediate proliferative and fibrogenic responses in HSCs.HSCs from WT, NOX1KO and NOX4KO mice were cultured for 1 day (quiescent HSCs) and 5 days (activated HSCs) with 10% FBS in DMEM. mRNA expression of proliferative genes (A) and fibrogenic genes (B) were measured by quantitative real-time PCR. HPRT was used as an internal control. *P <0.05, **P < 0.01, ***P < 0.001 vs activated WT HSCs control.
Mentions: We next investigated whether similar findings are observed in cultured HSCs treated with PDGF. As expected, the number of Ki67+HSCs isolated from WT was increased by PDGF treatment. However, PDGF produced significantly fewer Ki67+HSCs from NOX1KO and NOX4KO mice (Fig 6B and 6D). Furthermore, upregulation of proliferative genes (Cyclin D1, Bcl-2, PCNA and PDGFRB) was observed in activated WT HSCs (5 day plastic culture with 10% FBS medium). Proliferation of HSCs was attenuated by NOX1 and NOX4 deficiency. However, there was no change of proliferative genes in quiescent HSCs (1 day plastic culture) (Fig 7A). These results suggest that NOX1 and NOX4 mediate proliferation of HSCs in response to proliferative stimulation.

Bottom Line: Furthermore, the mRNAs of proliferative and pro-fibrotic genes were downregulated in NOX1 and NOX4 knock-out activated HSCs (cultured on plastic for 5 days).Finally, NOX1 and NOX4 protein levels were increased in human livers with cirrhosis compared with normal controls.Thus, NOX1 and NOX4 signaling mediates the pathogenesis of liver fibrosis, including the direct activation of HSC.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of California San Diego, La Jolla, California, United States of America; Vascular Biology Research Institute, Guangdong Pharmaceutical University, Guangzhou, China.

ABSTRACT
Reactive oxygen species (ROS) produced by nicotinamide adenine dinucleotide phosphate oxidase (NOX) play a key role in liver injury and fibrosis. Previous studies demonstrated that GKT137831, a dual NOX1/4 inhibitor, attenuated liver fibrosis in mice as well as pro-fibrotic genes in hepatic stellate cells (HSCs) as well as hepatocyte apoptosis. The effect of NOX1 and NOX4 deficiency in liver fibrosis is unclear, and has never been directly compared. HSCs are the primary myofibroblasts in the pathogenesis of liver fibrosis. Therefore, we aimed to determine the role of NOX1 and NOX4 in liver fibrosis, and investigated whether NOX1 and NOX4 signaling mediates liver fibrosis by regulating HSC activation. Mice were treated with carbon tetrachloride (CCl4) to induce liver fibrosis. Deficiency of either NOX1 or NOX4 attenuates liver injury, inflammation, and fibrosis after CCl4 compared to wild-type mice. NOX1 or NOX4 deficiency reduced lipid peroxidation and ROS production in mice with liver fibrosis. NOX1 and NOX4 deficiency are approximately equally effective in preventing liver injury in the mice. The NOX1/4 dual inhibitor GKT137831 suppressed ROS production as well as inflammatory and proliferative genes induced by lipopolysaccharide (LPS), platelet-derived growth factor (PDGF), or sonic hedgehog (Shh) in primary mouse HSCs. Furthermore, the mRNAs of proliferative and pro-fibrotic genes were downregulated in NOX1 and NOX4 knock-out activated HSCs (cultured on plastic for 5 days). Finally, NOX1 and NOX4 protein levels were increased in human livers with cirrhosis compared with normal controls. Thus, NOX1 and NOX4 signaling mediates the pathogenesis of liver fibrosis, including the direct activation of HSC.

No MeSH data available.


Related in: MedlinePlus