Ac2‑26 alleviates hepatic ischemia‑reperfusion injury based on inhibiting the positive feedback loop of HMGB1/TLR4/NF‑κB/neutrophils

Bai,Chen; Jiang,Zhengchen; Jiang,Hongxin; Yu,Shuna; Li,Ming; Chu,Fangfang; Sheng,Yaxin; Li,Jianguo; Jiang,Jiying; Li,Wanzhen

doi:10.3892/etm.2022.11609

Ac2‑26 alleviates hepatic ischemia‑reperfusion injury based on inhibiting the positive feedback loop of HMGB1/TLR4/NF‑κB/neutrophils

Authors:
- Chen Bai
- Zhengchen Jiang
- Hongxin Jiang
- Shuna Yu
- Ming Li
- Fangfang Chu
- Yaxin Sheng
- Jianguo Li
- Jiying Jiang
- Wanzhen Li
View Affiliations

Affiliations: Department of Anatomy, Basic Medical College of Weifang Medical University, Weifang, Shandong 261053, P.R. China, General Surgery Department, Shandong Provincial Qianfoshan Hospital, Jinan, Shandong 250013, P.R. China, Morphology Laboratory, Basic Medical College of Weifang Medical University, Weifang, Shandong 261053, P.R. China
Published online on: September 14, 2022 https://doi.org/10.3892/etm.2022.11609
Article Number: 673
Copyright: © Bai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Inflammation is one of the most crucial mechanism underlying hepatic ischemia‑reperfusion injury (HIRI). Several studies have shown that Ac2‑26, the active N‑terminal peptide of Annexin A1, could modulate anti‑inflammatory processes and protect the organs from ischemia‑reperfusion injury (IRI). However the effects of Ac2‑26 on an HIRI model have not been reported to date. The purpose of the present study was to determine whether Ac2‑26 pretreatment could protect hepatocytes against acute HIRI by inhibiting neutrophil infiltration through regulation of the high mobility group box protein 1 (HMGB1)/Toll‑like receptor 4 (TLR4)/NF‑κB signaling pathway. To this end, a total of 72 adult C57BL/6 mice were randomly divided into sham operation (sham), ischemia‑reperfusion (I/R), I/R + Ac2‑26 and Ac2‑26 groups. The HIRI model was established by occluding the branch of the hepatic pedicle to the left and median liver lobes with an atraumatic vascular clamp for 45 min, followed by reperfusion for 24 h. The expression of HMGB1, TLR4, NF‑κB, IκBα and lymphocyte antigen 6 complex locus G6D (Ly6G) was detected using reverse transcription‑quantitative PCR, western blotting and immunohistochemical staining; serum levels of HMGB1 were evaluated using an enzyme‑linked immunosorbent assay. Flow cytometry was used to detect the proportion of neutrophil. The results indicated that Ac2‑26 preconditioning rescued hepatocyte dysfunctions induced by HIRI. In addition, HIRI was associated with a significant increase in HMGB1 expression and release, accompanied by increased expression of TLR4, which was significantly inhibited by Ac2‑26. Furthermore, the expression of phosphorylated (p)‑NF‑κB and the ratio of p‑NF‑κB to NF‑κB were markedly increased, while the expression of IκBα was decreased in the I/R group compared with those in the sham group; however, these effects were reversed by Ac2‑26 administration. Additionally, Ac2‑26 administration significantly inhibited neutrophil infiltration and resulted in low levels of neutrophils and Ly6G as well as reduced myeloperoxidase activity. Taken together, these results indicated that Ac2‑26 pretreatment serves a protective role against HIRI by regulating the HMGB1/TLR4/NF‑κB signaling pathway and inhibiting neutrophil infiltration.

View Figures

View References

1	Saidi RF and Kenari SK: Liver ischemia/reperfusion injury: An overview. J Invest Surg. 6:366–379. 2014.PubMed/NCBI View Article : Google Scholar
2	van Golen RF, Reiniers MJ, Olthof PB, van Gulik TM and Heger M: Sterile inflammation in hepatic ischemia/reperfusion injury: Present concepts and potential therapeutics. J Gastroenterol Hepatol. 3:394–400. 2013.PubMed/NCBI View Article : Google Scholar
3	Quesnelle KM, Bystrom PV and Toledo-Pereyra LH: Molecular responses to ischemia and reperfusion in the liver. Arch Toxicol. 5:651–657. 2015.PubMed/NCBI View Article : Google Scholar
4	Wang Y, Liu ZS, Zhang SL, Diao QX and Ge YJ: Effect and mechanism of portal blood stasis removal on intestinal endotoxemia and hepatic ischemia reperfusion injury. Transplant Proc. 9:2752–2756. 2015.PubMed/NCBI View Article : Google Scholar
5	Huang H, Tohme S, Al-Khafaji AB, Tai S, Loughran P, Chen L, Wang S, Kim J, Billiar T, Wang Y and Tsung A: Damage-associated molecular pattern-activated neutrophil extracellular trap exacerbates sterile inflammatory liver injury. Hepatology. 2:600–614. 2015.PubMed/NCBI View Article : Google Scholar
6	Xu L, Ge F, Hu Y, Yu Y, Guo K and Miao C: Sevoflurane postconditioning attenuates hepatic ischemia-reperfusion injury by limiting HMGB1/TLR4/NF-kappaB pathway via modulating microRNA-142 in vivo and in vitro. Front Pharmacol. 12(646307)2021.PubMed/NCBI View Article : Google Scholar
7	Tsung A, Sahai R, Tanaka H, Nakao A, Fink MP, Lotze MT, Yang H, Li J, Tracey KJ, Geller DA and Billiar TR: The nuclear factor HMGB1 mediates hepatic injury after murine liver ischemia-reperfusion. J Exp Med. 7:1135–1143. 2005.PubMed/NCBI View Article : Google Scholar
8	Perretti M and D'Acquisto F: Annexin A1 and glucocorticoids as effectors of the resolution of inflammation. Nat Rev Immunol. 1:62–70. 2009.PubMed/NCBI View Article : Google Scholar
9	Yang YH, Morand E and Leech M: Annexin A1: Potential for glucocorticoid sparing in RA. Nat Rev Rheumatol. 10:595–603. 2013.PubMed/NCBI View Article : Google Scholar
10	La M, D'Amico M, Bandiera S, Di Filippo C, Oliani SM, Gavins FN, Flower RJ and Perretti M: Annexin 1 peptides protect against experimental myocardial ischemia-reperfusion: Analysis of their mechanism of action. FASEB J. 12:2247–2256. 2001.PubMed/NCBI View Article : Google Scholar
11	Guido BC, Zanatelli M, Tavares-De-Lima W, Oliani SM and Damazo AS: Annexin-A1 peptide down-regulates the leukocyte recruitment and up-regulates interleukin-10 release into lung after intestinal ischemia-reperfusion in mice. J Inflamm (Lond). 1(10)2013.PubMed/NCBI View Article : Google Scholar
12	Perretti M and Flower RJ: Annexin 1 and the biology of the neutrophil. J Leukoc Biol. 1:25–29. 2004.PubMed/NCBI View Article : Google Scholar
13	Facio FJ, Sena AA, Araujo LP, Mendes GE, Castro I, Luz MA, Yu L, Oliani SM and Burdmann EA: Annexin 1 mimetic peptide protects against renal ischemia/reperfusion injury in rats. J Mol Med (Berl). 1:51–63. 2011.PubMed/NCBI View Article : Google Scholar
14	Care NRCU, Animals AUOL: Guide for the care and use of laboratory animals. Washington (DC): National Academies Press (US), 2011.
15	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. 4:402–408. 2001.PubMed/NCBI View Article : Google Scholar
16	Wang J, Yu S, Li J, Li H, Jiang H, Xiao P, Pan Y, Zheng J, Yu L and Jiang J: Protective role of N-acetyl-l-tryptophan against hepatic ischemia-reperfusion injury via the RIP2/caspase-1/IL-1β signaling pathway. Pharm Biol. 1:385–391. 2019.PubMed/NCBI View Article : Google Scholar
17	Datta G, Fuller BJ and Davidson BR: Molecular mechanisms of liver ischemia reperfusion injury: Insights from transgenic knockout models. World J Gastroenterol. 11:1683–1698. 2013.PubMed/NCBI View Article : Google Scholar
18	Cirino G, Cicala C, Sorrentino L, Ciliberto G, Arpaia G, Perretti M and Flower RJ: Anti-inflammatory actions of an N-terminal peptide from human lipocortin 1. Br J Pharmacol. 3:573–574. 1993.PubMed/NCBI View Article : Google Scholar
19	Perretti M and Flower RJ: Modulation of IL-1-induced neutrophil migration by dexamethasone and lipocortin 1. J Immunol. 3:992–999. 1993.PubMed/NCBI
20	Perretti M, Ahluwalia A, Harris JG, Goulding NJ and Flower RJ: Lipocortin-1 fragments inhibit neutrophil accumulation and neutrophil-dependent edema in the mouse. A qualitative comparison with an anti-CD11b monoclonal antibody. J Immunol. 8:4306–4314. 1993.PubMed/NCBI
21	Kamaly N, Fredman G, Subramanian M, Gadde S, Pesic A, Cheung L, Fayad ZA, Langer R, Tabas I and Farokhzad OC: Development and in vivo efficacy of targeted polymeric inflammation-resolving nanoparticles. Proc Natl Acad Sci USA. 16:6506–6511. 2013.PubMed/NCBI View Article : Google Scholar
22	Gavins FN, Dalli J, Flower RJ, Granger DN and Perretti M: Activation of the annexin 1 counter-regulatory circuit affords protection in the mouse brain microcirculation. FASEB J. 8:1751–1758. 2007.PubMed/NCBI View Article : Google Scholar
23	Wang J, He GZ, Wang YK, Zhu QK, Chen W and Guo T: TLR4-HMGB1-, MyD88- and TRIF-dependent signaling in mouse intestinal ischemia/reperfusion injury. World J Gastroenterol. 27:8314–8325. 2015.PubMed/NCBI View Article : Google Scholar
24	Xue J, Ge H, Lin Z, Wang H, Lin W, Liu Y, Wu G, Xia J and Zhao Q: The role of dendritic cells regulated by HMGB1/TLR4 signalling pathway in myocardial ischaemia reperfusion injury. J Cell Mol Med. 4:2849–2862. 2019.PubMed/NCBI View Article : Google Scholar
25	Tsaroucha AK, Valsami G, Kostomitsopoulos N, Lambropoulou M, Anagnostopoulos C, Christodoulou E, Falidas E, Betsou A, Pitiakoudis M and Simopoulos CE: Silibinin effect on Fas/FasL, HMGB1, and CD45 expressions in a rat model subjected to liver ischemia-reperfusion injury. J Invest Surg. 6:491–502. 2018.PubMed/NCBI View Article : Google Scholar
26	Sugihara M, Sadamori H, Nishibori M, Sato Y, Tazawa H, Shinoura S, Umeda Y, Yoshida R, Nobuoka D, Utsumi M, et al: Anti-high mobility group box 1 monoclonal antibody improves ischemia/reperfusion injury and mode of liver regeneration after partial hepatectomy. Am J Surg. 1:179–188. 2016.PubMed/NCBI View Article : Google Scholar
27	Mcdonald KA, Huang H, Tohme S, Loughran P, Ferrero K, Billiar T and Tsung A: Toll-like receptor 4 (TLR4) antagonist eritoran tetrasodium attenuates liver ischemia and reperfusion injury through inhibition of high-mobility group box protein B1 (HMGB1) signaling. Mol Med. 20:639–648. 2015.PubMed/NCBI View Article : Google Scholar
28	Nace GW, Huang H, Klune JR, Eid RE, Rosborough BR, Korff S, Li S, Shapiro RA, Stolz DB, Sodhi CP, et al: Cellular-specific role of toll-like receptor 4 in hepatic ischemia-reperfusion injury in mice. Hepatology. 1:374–387. 2013.PubMed/NCBI View Article : Google Scholar
29	Zhu JR, Lu HD, Guo C, Fang WR, Zhao HD, Zhou JS, Wang F, Zhao YL, Li YM, Zhang YD, et al: Berberine attenuates ischemia-reperfusion injury through inhibiting HMGB1 release and NF-kappaB nuclear translocation. Acta Pharmacol Sin. 11:1706–1715. 2018.PubMed/NCBI View Article : Google Scholar
30	Zhang S, Feng Z, Gao W, Duan Y, Fan G, Geng X, Wu B, Li K, Liu K and Peng C: Aucubin attenuates liver ischemia-reperfusion injury by inhibiting the HMGB1/TLR-4/NF-kappaB signaling pathway, oxidative stress, and apoptosis. Front Pharmacol. 11(544124)2020.PubMed/NCBI View Article : Google Scholar
31	Leoni G, Neumann PA, Kamaly N, Quiros M, Nishio H, Jones HR, Sumagin R, Hilgarth RS, Alam A, Fredman G, et al: Annexin A1-containing extracellular vesicles and polymeric nanoparticles promote epithelial wound repair. J Clin Invest. 3:1215–1227. 2015.PubMed/NCBI View Article : Google Scholar
32	Reischl S, Lee JH, Miltschitzky J, Vieregge V, Walter RL, Twardy V, Kasajima A, Friess H, Kamaly N and Neumann PA: Ac2-26-nanoparticles induce resolution of intestinal inflammation and anastomotic healing via inhibition of NF-κB signaling in a model of perioperative colitis. Inflamm Bowel Dis. 9:1379–1393. 2021.PubMed/NCBI View Article : Google Scholar
33	Tsung A, Klune JR, Zhang X, Jeyabalan G, Cao Z, Peng X, Stolz DB, Geller DA, Rosengart MR and Billiar TR: HMGB1 release induced by liver ischemia involves Toll-like receptor 4 dependent reactive oxygen species production and calcium-mediated signaling. J Exp Med. 12:2913–2923. 2007.PubMed/NCBI View Article : Google Scholar
34	Baxter GF: The neutrophil as a mediator of myocardial ischemia-reperfusion injury: Time to move on. Basic Res Cardiol. 4:268–275. 2002.PubMed/NCBI View Article : Google Scholar
35	Vinten-Johansen J: Involvement of neutrophils in the pathogenesis of lethal myocardial reperfusion injury. Cardiovasc Res. 3:481–497. 2004.PubMed/NCBI View Article : Google Scholar
36	Yago T, Petrich BG, Zhang N, Liu Z, Shao B, Ginsberg MH and Mcever RP: Blocking neutrophil integrin activation prevents ischemia-reperfusion injury. J Exp Med. 8:1267–1281. 2015.PubMed/NCBI View Article : Google Scholar
37	Jaeschke H, Farhood A and Smith CW: Neutrophil-induced liver cell injury in endotoxin shock is a CD11b/CD18-dependent mechanism. Am J Physiol. 261 (6 Pt 1):G1051–G1056. 1991.PubMed/NCBI View Article : Google Scholar
38	Hoffman JJ, Gilbert TB, Poston RS and Silldorff EP: Myocardial reperfusion injury: Etiology, mechanisms, and therapies. J Extra Corpor Technol. 4:391–411. 2004.PubMed/NCBI
39	Saiman Y and Friedman SL: The role of chemokines in acute liver injury. Front Physiol. 3(213)2012.PubMed/NCBI View Article : Google Scholar
40	Monson KM, Dowlatshahi S and Crockett ET: CXC-chemokine regulation and neutrophil trafficking in hepatic ischemia-reperfusion injury in P-selectin/ICAM-1 deficient mice. J Inflamm (Lond). 4(11)2007.PubMed/NCBI View Article : Google Scholar
41	Perry BC, Soltys D, Toledo AH and Toledo-Pereyra LH: Tumor necrosis factor-α in liver ischemia/reperfusion injury. J Invest Surg. 4:178–188. 2011.PubMed/NCBI View Article : Google Scholar
42	Uchida Y, Freitas MC, Zhao D, Busuttil RW and Kupiec-Weglinski JW: The inhibition of neutrophil elastase ameliorates mouse liver damage due to ischemia and reperfusion. Liver Transpl. 8:939–947. 2009.PubMed/NCBI View Article : Google Scholar