Fluorofenidone attenuates pulmonary inflammation and fibrosis via inhibiting the activation of NALP3 inflammasome and IL-1β/IL-1R1/MyD88/NF-κB pathway.
Interleukin (IL)-1β plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. The production of IL-1β is dependent upon caspase-1-containing multiprotein complexes called inflammasomes and IL-1R1/MyD88/NF-κB pathway. In this study, we explored whether a potential anti-fibrotic agent fluorofenidone (FD) exerts its anti-inflammatory and anti-fibrotic effects through suppressing activation of NACHT, LRR and PYD domains-containing protein 3 (NALP3) inflammasome and the IL-1β/IL-1R1/MyD88/NF-κB pathway in vivo and in vitro. Male C57BL/6J mice were intratracheally injected with Bleomycin (BLM) or saline. Fluorofenidone was administered throughout the course of the experiment. Lung tissue sections were stained with haemotoxylin and eosin and Masson's trichrome. Cytokines were measured by ELISA, and α-smooth muscle actin (α-SMA), fibronectin, collagen I, caspase-1, IL-1R1, MyD88 were measured by Western blot and/or RT-PCR. The human actue monocytic leukaemia cell line (THP-1) were incubated with monosodium urate (MSU), with or without FD pre-treatment. The expression of caspase-1, IL-1β, NALP3, apoptosis-associated speck-like protein containing (ASC) and pro-caspase-1 were measured by Western blot, the reactive oxygen species (ROS) generation was detected using the Flow Cytometry, and the interaction of NALP3 inflammasome-associated molecules were measured by Co-immunoprecipitation. RLE-6TN (rat lung epithelial-T-antigen negative) cells were incubated with IL-1β, with or without FD pre-treatment. The expression of nuclear protein p65 was measured by Western blot. Results showed that FD markedly reduced the expressions of IL-1β, IL-6, monocyte chemotactic protein-1 (MCP-1), myeloperoxidase (MPO), α-SMA, fibronectin, collagen I, caspase-1, IL-1R1 and MyD88 in mice lung tissues. And FD inhibited MSU-induced the accumulation of ROS, blocked the interaction of NALP3 inflammasome-associated molecules, decreased the level of caspase-1 and IL-1β in THP-1 cells. Besides, FD inhibited IL-1β-induced the expression of nuclear protein p65. This study demonstrated that FD, attenuates BLM-induced pulmonary inflammation and fibrosis in mice via inhibiting the activation of NALP3 inflammasome and the IL-1β/IL-1R1/MyD88/ NF-κB pathway.
Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha, China.|Department of Respiratory Medicine, Central Hospital of Wuhan, Tongji Medical College Huazhong University of Science & Technology, Wuhan, China.
Cheng
C
Song
Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha, China.
Lujuan
L
He
Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha, China.
Jin
J
Zhang
Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha, China.
Hong
H
Ma
Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha, China.
Xiangning
X
Yuan
Pharmaceutical School, Central South University, Changsha, China.
Gaoyun
G
Hu
Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha, China.
Lijian
L
Tao
Department of Microbial Infection & Immunity, The Ohio State University, Columbus, OH, USA.
Jian
J
Zhang
Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha, China. mengjie@csu.edu.cn.
Jie
J
Meng
D000199:Actins / Q000378:metabolism; D000818:Animals; D001761:Bleomycin; D020170:Caspase 1 / Q000378:metabolism; D018932:Chemokine CCL2 / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B / Q000378:metabolism; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism*; D009195:Peroxidase / Q000378:metabolism; D011014:Pneumonia / Q000139:chemically induced / Q000150:complications* / Q000188:drug therapy* / Q000378:metabolism; D011658:Pulmonary Fibrosis / Q000150:complications* / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D011728:Pyridones / Q000494:pharmacology / Q000627:therapeutic use*; D017382:Reactive Oxygen Species / Q000378:metabolism; D018123:Receptors, Interleukin / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014527:Uric Acid / Q000494:pharmacology
C511655:5-methyl-1-(3-fluorophenyl)-2-(1H)-pyridone; C558396:Acta2 protein, mouse; D000199:Actins; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D024042:Collagen Type I; D005353:Fibronectins; D058847:Inflammasomes; D053583:Interleukin-1beta; D015850:Interleukin-6; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D011728:Pyridones; D017382:Reactive Oxygen Species; D018123:Receptors, Interleukin; D001761:Bleomycin; D014527:Uric Acid; D009195:Peroxidase; D020170:Caspase 1
IL-1β/IL-1R1/MyD88/NF-κB pathway; NALP3 inflammasome; fluorofenidone; pulmonary fibrosis