Pirfenidone Is an Agonistic Ligand for PPARα and Improves NASH by Activation of SIRT1/LKB1/pAMPK

Abstract

Nonalcoholic steatohepatitis (NASH) is recognized by hepatic lipid accumulation, inflammation, and fibrosis. No studies have evaluated the prolonged‐release pirfenidone (PR‐PFD) properties on NASH features. The aim of this study is to evaluate how PR‐PFD performs on metabolic functions, and provide insight on a mouse model of human NASH. Male C57BL/6J mice were fed with either normo diet or high‐fat/carbohydrate diet for 16 weeks and a subgroup also fed with PR‐PFD (300 mg/kg/day). An insulin tolerance test was performed at the end of treatment. Histological analysis, determination of serum hormones, adipocytokines measurement, and evaluation of proteins by western blot was performed. Molecular docking, in silico site‐directed mutagenesis, and in vitro experiments using HepG2 cultured cells were performed to validate PR‐PFD binding to peroxisome proliferator–activated receptor alpha (PPAR‐α), activation of PPAR‐α promoter, and sirtuin 1 (SIRT1) protein expression. Compared with the high‐fat group, the PR‐PFD‐treated mice displayed less weight gain, cholesterol, very low density lipoprotein and triglycerides, and showed a significant reduction of hepatic macrosteatosis, inflammation, hepatocyte ballooning, fibrosis, epididymal fat, and total adiposity. PR‐PFD restored levels of insulin, glucagon, adiponectin, and resistin along with improved insulin resistance. Noteworthy, SIRT1–liver kinase B1–phospho‐5′ adenosine monophosphate–activated protein kinase signaling and the PPAR‐α/carnitine O‐palmitoyltransferase 1/acyl‐CoA oxidase 1 pathway were clearly induced in high fat + PR‐PFD mice. In HepG2 cells incubated with palmitate, PR‐PFD induced activation and nuclear translocation of both PPARα and SIRT1, which correlated with increased SIRT1 phosphorylated in serine 47, suggesting a positive feedback loop between the two proteins. These results were confirmed with both synthetic PPAR‐α and SIRT1 activators and inhibitors. Finally, we found that PR‐PFD is a true agonist/ligand for PPAR‐α. Conclusions: PR‐PFD provided an anti‐steatogenic effect and protection for inflammation and fibrosis.

PPARα Agonist WY-14643 Relieves Neuropathic Pain through SIRT1-Mediated Deacetylation of NF-κB

“…It suggested that PPAR α /SIRT1 signaling might be relative to the inflammation progression of NP. The results were consistent with the clues that PPAR α had a hand in inflammation by regulating SIRT1 [ 28 ]. Moreover, since SIRT1 was capable of bating inflammation by inhibition of NF- κ B [ 15 , 17 ], we also determined the levels of SIRT1 and acetylated NF- κ B p65 in CCI rats.…”

“…Ogawa et al found that the repression of microglial activation was associated with SIRT1-dependent PPAR α signaling [ 27 ]. Sandoval-Rodriguez et al showed that PPAR α improved nonalcoholic steatohepatitis via acting on SIRT1 [ 28 ]. However, whether the PPAR α /SIRT1 pathway was involved in NP development was unclear.…”

Liver Cancer: Therapeutic Challenges and the Importance of Experimental Models

“…Additionally, PFD has shown to be a powerful antifibrotic drug at dose of 300 mg/kg in an experimental HCC model induced by carbon tetrachloride in mice [66], but the pharmacodynamic mechanisms involved in responses generated by PFD have yet to be clarified. It has recently been shown that pirfenidone can bind to ligand binding domain of PPARα, which is a PPARc homolog, activating the SIRT1/LκB1/pAMPK indicating their ability to modify the epigenetic marks of the H3K9 [67]. Table 2 summarizes some important pharmacological characteristics of each one of the previously described therapies.…”

Roles of Nrf2 in Liver Diseases: Molecular, Pharmacological, and Epigenetic Aspects

“…MAFLD includes NASH, fibrosis, cirrhosis, and hepatocellular carcinoma [ 26 , 27 ]. Clinical and experimental data suggest that fatty liver can not only induce serious tissue damage and even cancer, but can also cause cardiovascular diseases [ 28 , 29 ].…”