Rev‑erb&alpha;: The circadian guardian against NLRP3‑driven liver fibrosis

Wang,Junmin; Wang,Yanping; Lin,Liubing; Pei,Wen; Li,Yong

doi:10.3892/mmr.2025.13635

October-2025 Volume 32 Issue 4

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

October-2025 Volume 32 Issue 4

Full Size Image

Article Open Access

Rev‑erbα: The circadian guardian against NLRP3‑driven liver fibrosis

Authors:
- Junmin Wang
- Yanping Wang
- Liubing Lin
- Wen Pei
- Yong Li
View Affiliations / Copyright

Affiliations: Department of Gastroenterology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200071, P.R. China, Department of Cardiology, Jing'an District Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200072, P.R. China

Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Article Number: 270
|
Published online on: July 24, 2025

https://doi.org/10.3892/mmr.2025.13635
Expand metrics +

Abstract

Liver fibrosis is a pivotal pathological process in the progression of various chronic liver diseases toward cirrhosis, primarily driven by the activation of hepatic stellate cells. Recent studies have implicated dysregulation of circadian clock genes in the pathogenesis of hepatic disorders. The present investigation focused on the role of the circadian regulator nuclear receptor subfamily 1 group D member 1 (Rev‑erbα) in liver fibrosis and its mechanistic interplay with the NLR family domain containing protein 3 (NLRP3) inflammasome. A mouse model of liver fibrosis was established via carbon tetrachloride (CCl4) administration. The expression of Rev‑erbα was modulated pharmacologically using the agonist GSK4112 and the antagonist SR8278 to assess its impact on fibrogenesis. In parallel, lentiviral vectors were employed in in vitro studies to generate LX‑2 cell lines with Rev‑erbα overexpression or knockout. Transforming growth factor‑β1 (TGF‑β1) was applied to induce cellular activation, and subsequent effects on the NLRP3 inflammasome and its downstream mediators were analyzed. The extent of fibrosis and molecular alterations were evaluated using Masson's trichrome staining, Sirius Red staining, immunohistochemistry, western blotting and reverse transcription‑quantitative PCR. Rev‑erbα expression was significantly downregulated in both CCl4‑induced murine models and TGF‑β1‑activated LX‑2 cells. Pharmacological activation of Rev‑erbα attenuated hepatic fibrosis, evidenced by reduced collagen accumulation and suppression of fibrogenic markers (α‑smooth muscle actin, collagen 1 and TGF‑β1). By contrast, inhibition of Rev‑erbα exacerbated fibrotic responses. Mechanistically, Rev‑erbα activation inhibited NLRP3 inflammasome signaling and downstream pro‑inflammatory cytokines [interleukin (IL)‑18 and IL‑1β], underscoring its anti‑fibrotic function via NLRP3 pathway modulation. Rev‑erbα functions as a key negative regulator of hepatic fibrosis by suppressing NLRP3 inflammasome activation, representing a promising therapeutic target for the management of liver fibrosis.

View Figures

View References

1	Hernandez-Gea V and Friedman SL: Pathogenesis of liver fibrosis. Annu Rev Pathol. 6:425–456. 2011. View Article : Google Scholar : PubMed/NCBI
2	Parola M and Pinzani M: Liver fibrosis: Pathophysiology, pathogenetic targets and clinical issues. Mol Aspects Med. 65:37–55. 2019. View Article : Google Scholar : PubMed/NCBI
3	Higashi T, Friedman SL and Hoshida Y: Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev. 121:27–42. 2017. View Article : Google Scholar : PubMed/NCBI
4	Tsuchida T and Friedman SL: Mechanisms of hepatic stellate cell activation. Nat Rev Gastroenterol Hepatol. 14:397–411. 2017. View Article : Google Scholar : PubMed/NCBI
5	Frazier K, Manzoor S, Carroll K, DeLeon O, Miyoshi S, Miyoshi J, St George M, Tan A, Chrisler EA, Izumo M, et al: Gut microbes and the liver circadian clock partition glucose and lipid metabolism. J Clin Invest. 133:e1625152023. View Article : Google Scholar : PubMed/NCBI
6	Mukherji A, Bailey SM, Staels B and Baumert TF: The circadian clock and liver function in health and disease. J Hepatol. 71:200–211. 2019. View Article : Google Scholar : PubMed/NCBI
7	Piggins HD: Human clock genes. Ann Med. 34:394–400. 2002. View Article : Google Scholar : PubMed/NCBI
8	Cox KH and Takahashi JS: Circadian clock genes and the transcriptional architecture of the clock mechanism. J Mol Endocrinol. 63:R93–R102. 2019. View Article : Google Scholar : PubMed/NCBI
9	Tsang AH, Sánchez-Moreno C, Bode B, Rossner MJ, Garaulet M and Oster H: Tissue-specific interaction of Per1/2 and Dec2 in the regulation of fibroblast circadian rhythms. J Biol Rhythms. 27:478–489. 2012. View Article : Google Scholar : PubMed/NCBI
10	González-Fernández B, Sánchez DI, Crespo I, San-Miguel B, de Urbina JO, González-Gallego J and Tuñón MJ: Melatonin attenuates dysregulation of the circadian clock pathway in mice with CCl4-Induced fibrosis and human hepatic stellate cells. Front Pharmacol. 9:5562018. View Article : Google Scholar : PubMed/NCBI
11	Chen P, Kakan X, Wang S, Dong W, Jia A, Cai C and Zhang J: Deletion of clock gene Per2 exacerbates cholestatic liver injury and fibrosis in mice. Exp Toxicol Pathol. 65:427–432. 2013. View Article : Google Scholar : PubMed/NCBI
12	Chen P, Han Z, Yang P, Zhu L, Hua Z and Zhang J: Loss of clock gene mPer2 promotes liver fibrosis induced by carbon tetrachloride. Hepatol Res. 40:1117–1127. 2010. View Article : Google Scholar : PubMed/NCBI
13	Laitinen S, Fontaine C, Fruchart JC and Staels B: The role of the orphan nuclear receptor Rev-Erb alpha in adipocyte differentiation and function. Biochimie. 87:21–25. 2005. View Article : Google Scholar : PubMed/NCBI
14	Cho H, Zhao X, Hatori M, Yu RT, Barish GD, Lam MT, Chong LW, DiTacchio L, Atkins AR, Glass CK, et al: Regulation of circadian behaviour and metabolism by REV-ERB-α and REV-ERB-β. Nature. 485:123–127. 2012. View Article : Google Scholar : PubMed/NCBI
15	Stujanna EN, Murakoshi N, Tajiri K, Xu D, Kimura T, Qin R, Feng D, Yonebayashi S, Ogura Y, Yamagami F, et al: Rev-erb agonist improves adverse cardiac remodeling and survival in myocardial infarction through an anti-inflammatory mechanism. PLoS One. 12:e01893302017. View Article : Google Scholar : PubMed/NCBI
16	Ding G, Li X, Hou X, Zhou W, Gong Y, Liu F, He Y, Song J, Wang J, Basil P, et al: REV-ERB in GABAergic neurons controls diurnal hepatic insulin sensitivity. Nature. 592:763–767. 2021. View Article : Google Scholar : PubMed/NCBI
17	da Rocha AL, Pinto AP, Bedo BLS, Morais GP, Oliveira LC, Carolino ROG, Pauli JR, Simabuco FM, de Moura LP, Ropelle ER, et al: Exercise alters the circadian rhythm of REV-ERB-α and downregulates autophagy-related genes in peripheral and central tissues. Sci Rep. 12:200062022. View Article : Google Scholar : PubMed/NCBI
18	Luo X, Song S, Qi L, Tien CL, Li H, Xu W, Mathuram TL, Burris T, Zhao Y, Sun Z and Zhang L: REV-ERB is essential in cardiac fibroblasts homeostasis. Front Pharmacol. 13:8996282022. View Article : Google Scholar : PubMed/NCBI
19	Griffin P, Dimitry JM and Musiek ES: Rev-erbs and Glia-implications for neurodegenerative diseases. J Exp Neurosci. 13:11790695198532332019. View Article : Google Scholar : PubMed/NCBI
20	Hu C, Zhao L, Tao J and Li L: Protective role of melatonin in Early-stage and end-stage liver cirrhosis. J Cell Mol Med. 23:7151–7162. 2019. View Article : Google Scholar : PubMed/NCBI
21	Chen L, Xia S, Wang F, Zhou Y, Wang S, Yang T, Li Y, Xu M, Zhou Y, Kong D, et al: m6A methylation-induced NR1D1 ablation disrupts the HSC circadian clock and promotes hepatic fibrosis hepatic fibrosis. Pharmacol Res. 189:1067042023. View Article : Google Scholar : PubMed/NCBI
22	Wang S, Lin Y, Yuan X, Li F, Guo L and Wu B: REV-ERBα integrates colon clock with experimental colitis through regulation of NF-κB/NLRP3 axis. Nat Commun. 9:42462018. View Article : Google Scholar : PubMed/NCBI
23	Tao Y, Wang N, Qiu T and Sun X: The Role of Autophagy and NLRP3 inflammasome in liver fibrosis. Biomed Res Int. 2020:72691502020. View Article : Google Scholar : PubMed/NCBI
24	Wree A, McGeough MD, Inzaugarat ME, Eguchi A, Schuster S, Johnson CD, Peña CA, Geisler LJ, Papouchado BG, Hoffman HM, et al: NLRP3 inflammasome driven liver injury and fibrosis: Roles of IL-17 and TNF in mice. Hepatology. 67:736–749. 2018. View Article : Google Scholar : PubMed/NCBI
25	Zhang K, Lin L, Zhu Y, Zhang N, Zhou M and Li Y: Saikosaponin d alleviates liver fibrosis by negatively regulating the ROS/NLRP3 inflammasome through activating the ERβ pathway. Front Pharmacol. 13:8949812022. View Article : Google Scholar : PubMed/NCBI
26	Shao R, Yang Y, Fan K, Wu X, Jiang R, Tang L, Li L, Shen Y, Liu G and Zhang L: REV-ERBα Agonist GSK4112 attenuates Fas-induced Acute Hepatic Damage in Mice. Int J Med Sci. 18:3831–3838. 2021. View Article : Google Scholar : PubMed/NCBI
27	Kim J, Park I, Jang S, Choi M, Kim D, Sun W, Choe Y, Choi JW, Moon C, Park SH, et al: Pharmacological rescue with SR8278, a circadian nuclear receptor REV-ERBα antagonist as a therapy for mood disorders in Parkinson's disease. Neurotherapeutics. 19:592–607. 2022. View Article : Google Scholar : PubMed/NCBI
28	Kojetin D, Wang Y, Kamenecka TM and Burris TP: Identification of SR8278, a synthetic antagonist of the nuclear heme receptor REV-ERB. ACS Chem Biol. 6:131–134. 2011. View Article : Google Scholar : PubMed/NCBI
29	Lin L, Zhou M, Que R, Chen Y, Liu X, Zhang K, Shi Z and Li Y: Saikosaponin-d protects against liver fibrosis by regulating the estrogen receptor-β/NLRP3 inflammasome pathway. Biochem Cell Biol. 99:666–674. 2021. View Article : Google Scholar : PubMed/NCBI
30	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
31	Yue J, He J, Wei Y, Shen K, Wu K, Yang X, Liu S, Zhang C and Yang H: Decreased expression of Rev-Erbα in the epileptic foci of temporal lobe epilepsy and activation of Rev-Erbα have anti-inflammatory and neuroprotective effects in the pilocarpine model. J Neuroinflammation. 17:432020. View Article : Google Scholar : PubMed/NCBI
32	Niu WX, Bao YY, Zhang N, Lu ZN, Ge MX, Li YM, Li Y, Chen MH and He HW: Dehydromevalonolactone ameliorates liver fibrosis and inflammation by repressing activation of NLRP3 inflammasome. Bioorg Chem. 127:1059712022. View Article : Google Scholar : PubMed/NCBI
33	Zhang X, Kuang G, Wan J, Jiang R, Ma L, Gong X and Liu X: Salidroside protects mice against CCl4-induced acute liver injury via down-regulating CYP2E1 expression and inhibiting NLRP3 inflammasome activation. Int Immunopharmacol. 85:1066622020. View Article : Google Scholar : PubMed/NCBI
34	Kisseleva T and Brenner D: Molecular and cellular mechanisms of liver fibrosis and its regression. Nat Rev Gastroenterol Hepatol. 18:151–166. 2021. View Article : Google Scholar : PubMed/NCBI
35	Pan X, Mota S and Zhang B: Circadian clock regulation on lipid metabolism and metabolic diseases. Adv Exp Med Biol. 1276:53–66. 2020. View Article : Google Scholar : PubMed/NCBI
36	Bailey SM: Emerging role of circadian clock disruption in Alcohol-induced liver disease. Am J Physiol Gastrointest Liver Physiol. 315:G364–G373. 2018. View Article : Google Scholar : PubMed/NCBI
37	Richards J and Gumz ML: Advances in understanding the peripheral circadian clocks. FASEB J. 26:3602–3613. 2012. View Article : Google Scholar : PubMed/NCBI
38	Horii R, Honda M, Shirasaki T, Shimakami T, Shimizu R, Yamanaka S, Murai K, Kawaguchi K, Arai K, Yamashita T, et al: MicroRNA-10a impairs liver metabolism in Hepatitis C Virus-related cirrhosis through deregulation of the circadian clock gene brain and Muscle Aryl hydrocarbon receptor nuclear Translocator-Like 1. Hepatol Commun. 3:1687–1703. 2019. View Article : Google Scholar : PubMed/NCBI
39	Zhang D, Tong X, Nelson BB, Jin E, Sit J, Charney N, Yang M, Omary MB and Yin L: The hepatic BMAL1/AKT/lipogenesis axis protects against alcoholic liver disease in mice via promoting PPARα pathway. Hepatology. 68:883–896. 2018. View Article : Google Scholar : PubMed/NCBI
40	Pan X, Queiroz J and Hussain MM: Nonalcoholic fatty liver disease in CLOCK mutant mice. J Clin Invest. 130:4282–4300. 2020.PubMed/NCBI
41	Zhou B, Zhang Y, Zhang F, Xia Y, Liu J, Huang R, Wang Y, Hu Y, Wu J, Dai C, et al: CLOCK/BMAL1 regulates circadian change of mouse hepatic insulin sensitivity by SIRT1. Hepatology. 59:2196–2206. 2014. View Article : Google Scholar : PubMed/NCBI
42	Gnocchi D, Custodero C, Sabbà C and Mazzocca A: Circadian rhythms: A possible new player in Non-alcoholic fatty liver disease pathophysiology. J Mol Med (Berl). 97:741–759. 2019. View Article : Google Scholar : PubMed/NCBI
43	Forman BM, Chen J, Blumberg B, Kliewer SA, Henshaw R, Ong ES and Evans RM: Cross-talk among ROR alpha 1 and the Rev-erb family of orphan nuclear receptors. Mol Endocrinol. 8:1253–1261. 1994. View Article : Google Scholar : PubMed/NCBI
44	Le Martelot G, Claudel T, Gatfield D, Schaad O, Kornmann B, Lo Sasso G, Moschetta A and Schibler U: REV-ERBalpha participates in circadian SREBP signaling and bile acid homeostasis. PLoS Biol. 7:e10001812009. View Article : Google Scholar : PubMed/NCBI
45	Zhou D, Wang Y, Chen L, Jia L, Yuan J, Sun M, Zhang W, Wang P, Zuo J, Xu Z and Luan J: Evolving roles of circadian rhythms in liver homeostasis and pathology. Oncotarget. 7:8625–8639. 2016. View Article : Google Scholar : PubMed/NCBI
46	Ni Y, Zhao Y, Ma L, Wang Z, Ni L, Hu L and Fu Z: Pharmacological activation of REV-ERBα improves nonalcoholic steatohepatitis by regulating intestinal permeability. Metabolism. 114:1544092021. View Article : Google Scholar : PubMed/NCBI
47	Lin Y, Wang S, Gao L, Zhou Z, Yang Z, Lin J, Ren S, Xing H and Wu B: Oscillating lncRNA Platr4 regulates NLRP3 inflammasome to ameliorate nonalcoholic steatohepatitis in mice. Theranostics. 11:426–444. 2021. View Article : Google Scholar : PubMed/NCBI
48	Yu F, Wang Z, Zhang T, Chen X, Xu H, Wang F, Guo L, Chen M, Liu K and Wu B: Deficiency of intestinal Bmal1 prevents obesity induced by high-fat feeding. Nat Commun. 12:53232021. View Article : Google Scholar : PubMed/NCBI
49	Lin Y, Lin L, Gao L, Wang S and Wu B: Rev-erbα regulates hepatic ischemia-reperfusion injury in mice. Biochem Biophys Res Commun. 529:916–921. 2020. View Article : Google Scholar : PubMed/NCBI
50	Liu Q, Xu L, Wu M, Zhou Y, Yang J, Huang C, Xu T, Li J and Zhang L: Rev-erbα exacerbates hepatic steatosis in alcoholic liver diseases through regulating autophagy. Cell Biosci. 11:1292021. View Article : Google Scholar : PubMed/NCBI
51	Yang Z, Smalling RV, Huang Y, Jiang Y, Kusumanchi P, Bogaert W, Wang L, Delker DA, Skill NJ, Han S, et al: The role of SHP/REV-ERBα/CYP4A axis in the pathogenesis of Alcohol-associated liver disease. JCI Insight. 6:e1406872021. View Article : Google Scholar : PubMed/NCBI
52	Wang J, Yin L and Lazar MA: The orphan nuclear receptor Rev-erb alpha regulates circadian expression of plasminogen activator inhibitor type 1. J Biol Chem. 281:33842–33848. 2006. View Article : Google Scholar : PubMed/NCBI
53	Crouchet E, Dachraoui M, Jühling F, Roehlen N, Oudot MA, Durand SC, Ponsolles C, Gadenne C, Meiss-Heydmann L, Moehlin J, et al: Targeting the liver clock improves fibrosis by restoring TGF-β signaling. J Hepatol. 82:120–133. 2025. View Article : Google Scholar : PubMed/NCBI
54	Grant D, Yin L, Collins JL, Parks DJ, Orband-Miller LA, Wisely GB, Joshi S, Lazar MA, Willson TM and Zuercher WJ: GSK4112, a small molecule chemical probe for the cell biology of the nuclear heme receptor Rev-erbα. ACS Chem Biol. 5:925–932. 2010. View Article : Google Scholar : PubMed/NCBI
55	Dong D, Sun H, Wu Z, Wu B, Xue Y and Li Z: A validated ultra-performance liquid chromatography-tandem mass spectrometry method to identify the pharmacokinetics of SR8278 in normal and streptozotocin-induced diabetic rats. J Chromatogr B Analyt Technol Biomed Life Sci. 1020:142–147. 2016. View Article : Google Scholar : PubMed/NCBI
56	Ishizuka T and Lazar MA: The N-CoR/histone deacetylase 3 complex is required for repression by thyroid hormone receptor. Mol Cell Biol. 23:5122–5131. 2003. View Article : Google Scholar : PubMed/NCBI
57	Yin L and Lazar MA: The orphan nuclear receptor Rev-erbalpha recruits the N-CoR/histone deacetylase 3 corepressor to regulate the circadian Bmal1 gene. Mol Endocrinol. 19:1452–1459. 2005. View Article : Google Scholar : PubMed/NCBI
58	Yin L, Joshi S, Wu N, Tong X and Lazar MA: E3 ligases Arf-bp1 and Pam mediate lithium-stimulated degradation of the circadian heme receptor Rev-erb alpha. Proc Natl Acad Sci USA. 107:11614–11619. 2010. View Article : Google Scholar : PubMed/NCBI
59	Yin L, Wang J, Klein PS and Lazar MA: Nuclear receptor Rev-erbalpha is a critical lithium-sensitive component of the circadian clock. Science. 311:1002–1005. 2006. View Article : Google Scholar : PubMed/NCBI
60	Kisseleva T and Brenner DA: Hepatic stellate cells and the reversal of fibrosis. J Gastroenterol Hepatol. 21 (Suppl 3):S84–S87. 2006. View Article : Google Scholar : PubMed/NCBI
61	Liu XY, Liu RX, Hou F, Cui LJ, Li CY, Chi C, Yi E, Wen Y and Yin CH: Fibronectin expression is critical for liver fibrogenesis in vivo and in vitro. Mol Med Rep. 14:3669–3675. 2016. View Article : Google Scholar : PubMed/NCBI
62	Friedman SL: Hepatic stellate cells: Protean, multifunctional, and enigmatic cells of the liver. Physiol Rev. 88:125–172. 2008. View Article : Google Scholar : PubMed/NCBI
63	Xiang D, Zou J, Zhu X, Chen X, Luo J, Kong L and Zhang H: Physalin D attenuates hepatic stellate cell activation and liver fibrosis by blocking TGF-β/Smad and YAP signaling. Phytomedicine. 78:1532942020. View Article : Google Scholar : PubMed/NCBI
64	Zhang J, Jiang N, Ping J and Xu L: TGF-β1-induced autophagy activates hepatic stellate cells via the ERK and JNK signaling pathways. Int J Mol Med. 47:256–266. 2021. View Article : Google Scholar : PubMed/NCBI
65	Wu Z, Liao F, Luo G, Qian Y, He X, Xu W, Ding S and Pu J: NR1D1 Deletion induces Rupture-prone vulnerable plaques by regulating macrophage pyroptosis via the NF-κB/NLRP3 inflammasome pathway. Oxid Med Cell Longev. 2021:52175722021. View Article : Google Scholar : PubMed/NCBI
66	Ka NL, Park MK, Kim SS, Jeon Y, Hwang S, Kim SM, Lim GY, Lee H and Lee MO: NR1D1 Stimulates antitumor immune responses in breast cancer by activating cGAS-STING signaling. Cancer Res. 83:3045–3058. 2023. View Article : Google Scholar : PubMed/NCBI
67	Kou L, Chi X, Sun Y, Han C, Wan F, Hu J, Yin S, Wu J, Li Y, Zhou Q, et al: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson's disease via the microglial NLRP3 inflammasome. J Neuroinflammation. 19:1332022. View Article : Google Scholar : PubMed/NCBI
68	Huang ZN, Wang J, Wang ZY, Min LY, Ni HL, Han YL, Tian YY, Cui YZ, Han JX and Cheng XF: SR9009 attenuates inflammation-related NPMSC pyroptosis and IVDD through NR1D1/NLRP3/IL-1β pathway. iScience. 27:1097332024. View Article : Google Scholar : PubMed/NCBI