Fluorofenidone protects against acute liver failure in mice by regulating MKK4/JNK pathway

Lei Gu; Xin He; Yanqiu Zhang; Shenglan Li; Jie Tang; Ruixue Ma; Xinyi Yang; Hao Huang; Yu Peng; Yanyun Xie; Zhangzhe Peng; Jie Meng; Gaoyun Hu; Lijian Tao; Xiaowei Liu; Huixiang Yang

doi:10.1016/j.biopha.2023.114844

Fluorofenidone protects against acute liver failure in mice by regulating MKK4/JNK pathway

Biomed Pharmacother. 2023 Aug:164:114844. doi: 10.1016/j.biopha.2023.114844. Epub 2023 May 22.

Authors

Lei Gu¹, Xin He², Yanqiu Zhang³, Shenglan Li³, Jie Tang⁴, Ruixue Ma⁴, Xinyi Yang⁴, Hao Huang⁵, Yu Peng⁴, Yanyun Xie⁶, Zhangzhe Peng⁷, Jie Meng⁸, Gaoyun Hu⁹, Lijian Tao¹⁰, Xiaowei Liu¹¹, Huixiang Yang¹²

Affiliations

¹ Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China.
² Department of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China.
³ Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China.
⁴ Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China.
⁵ Department of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; Department of Cell Biology, School of Life Sciences, Central South University, Changsha 410013, China.
⁶ Department of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China.
⁷ Department of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China. Electronic address: pengzhangzhe@csu.edu.cn.
⁸ Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China; Department of Respirology, Third Xiangya Hospital, Central South University, Changsha 410013, China.
⁹ Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; Faculty of Pharmaceutical Sciences, Central South University, Changsha 410013, China.
¹⁰ Department of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China.
¹¹ Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China.
¹² Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China; Hunan Key Lab of Organ Fibrosis, Changsha 410008, China; National International Collaborative Research Center for Medical Metabolomics, Xiangya Hospital, Central South University, Changsha 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China. Electronic address: yang_hx430@csu.edu.cn.

PMID: 37224750
DOI: 10.1016/j.biopha.2023.114844

Abstract

Aims: Acute liver failure (ALF) is a life-threatening disease characterized by abrupt and extensive hepatic necrosis and apoptosis, resulting in high mortality. The approved drug, N-acetylcysteine (NAC), is only effective for acetaminophen (APAP)-associated ALF at the early stage. Thus, we investigate whether fluorofenidone (AKF-PD), a novel antifibrosis pyridone agent, protects against ALF in mice and explore its underlying mechanisms.

Methods: ALF mouse models were established using APAP or lipopolysaccharide/D-galactosamine (LPS/D-Gal). Anisomycin and SP600125 were used as JNK activator and inhibitor, respectively, and NAC served as a positive control. Mouse hepatic cell line AML12 and primary mouse hepatocytes were used for in vitro studies.

Results: AKF-PD pretreatment alleviated APAP-induced ALF with decreased necrosis, apoptosis, reactive oxygen species (ROS) markers, and mitochondrial permeability transition in liver. Additionally, AKF-PD alleviated mitochondrial ROS stimulated by APAP in AML12 cells. RNA-sequencing in the liver and subsequent gene set enrichment analysis showed that AKF-PD significantly impacted MAPK and IL-17 pathway. In vitro and in vivo studies demonstrated that AKF-PD inhibited APAP-induced phosphorylation of MKK4/JNK, while SP600125 only inhibited JNK phosphorylation. The protective effect of AKF-PD was abolished by anisomycin. Similarly, AKF-PD pretreatment abolished hepatotoxicity caused by LPS/D-Gal, decreased ROS levels, and diminished inflammation. Furthermore, unlike NAC, AKF-PD, inhibited the phosphorylation of MKK4 and JNK upon pretreatment, and improved survival in cases of LPS/D-Gal-induced mortality with delayed dosing.

Conclusions: In summary, AKF-PD can protect against ALF caused by APAP or LPS/D-Gal, in part, via regulating MKK4/JNK pathway. AKF-PD might be a novel candidate drug for ALF.

Keywords: Acute liver failure; Fluorofenidone; JNK; MKK4; N-acetylcysteine.

MeSH terms

Acetaminophen / metabolism
Animals
Anisomycin / metabolism
Anisomycin / pharmacology
Hepatocytes
Lipopolysaccharides / pharmacology
Liver
Liver Failure, Acute* / chemically induced
Liver Failure, Acute* / drug therapy
Liver Failure, Acute* / prevention & control
MAP Kinase Signaling System*
Mice
Mice, Inbred C57BL
Necrosis / metabolism
Pyridones / pharmacology
Reactive Oxygen Species / metabolism

Substances

pyrazolanthrone
5-methyl-1-(3-fluorophenyl)-2-(1H)-pyridone
Reactive Oxygen Species
Acetaminophen
Lipopolysaccharides
Anisomycin
Pyridones