BML-111 suppresses TGF-β1-induced lung ﬁbroblast activation in vitro and decreases experimental pulmonary fibrosis in vivo

Ji,Yu‑Dong; Luo,Zhen‑Long; Chen,Chun‑Xiu; Li,Bo; Gong,Jie; Wang,Ya‑Xin; Chen,Lin; Yao,Shang‑Long; Shang,You

doi:10.3892/ijmm.2018.3914

BML-111 suppresses TGF-β1-induced lung ﬁbroblast activation in vitro and decreases experimental pulmonary fibrosis in vivo

Authors:
- Yu‑Dong Ji
- Zhen‑Long Luo
- Chun‑Xiu Chen
- Bo Li
- Jie Gong
- Ya‑Xin Wang
- Lin Chen
- Shang‑Long Yao
- You Shang
View Affiliations

Affiliations: Department of Anesthesiology, Institute of Anesthesiology and Critical Care, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Critical Care Medicine, Institute of Anesthesiology and Critical Care, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
Published online on: October 2, 2018 https://doi.org/10.3892/ijmm.2018.3914
Pages: 3083-3092
Copyright: © Ji 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

Pulmonary fibrosis is an aggressive end‑stage disease. Transforming growth factor‑β1 (TGF‑β1) mediates lung ﬁbroblast activation and is essential for the progress of pulmonary fibrosis. BML‑111, a lipoxinA4 (LXA4) receptor (ALX) agonist, has been reported to possess anti‑ﬁbrotic properties. The present study aimed to elucidate whether BML‑111 inhibits TGF‑β1‑induced mouse embryo lung ﬁbroblast (NIH3T3 cell line) activation in vitro and bleomycin (BLM)‑induced pulmonary fibrosis in vivo. In vitro experiments demonstrated that BML‑111 treatment inhibits TGF‑β1‑induced NIH3T3 cell viability and the expression of smooth muscle α actin (α‑SMA), ﬁbronectin and total collagen. Furthermore, this suppressive effect was associated with mothers against decapentaplegic homolog (Smad)2/3, extracellular signal‑regulated kinase (ERK) and Akt phosphorylation interference. In vivo experiments revealed that BML‑111 treatment markedly improved survival rate and ameliorated the destruction of lung tissue structure. It also reduced interleukin‑1β (IL‑1β), tumor necrosis factor‑α (TNF‑α) and TGF‑β1 expression in the BLM intratracheal mouse model. In addition, the expression ofα‑SMA and extracellular matrix (ECM) deposition (total collagen, hydroxyproline and ﬁbronectin) were also suppressed following BML‑111 treatment. However, BOC‑2, an antagonist of ALX, partially weakened the effects of BML‑111. In conclusion, these results indicated that BML‑111 inhibits TGF‑β1‑induced ﬁbroblasts activation and alleviates BLM‑induced pulmonary fibrosis. Therefore, BML‑111 may be used as a potential therapeutic agent for pulmonary fibrosis treatment.

View Figures

View References

1	King TJ Jr, Tooze JA, Schwarz MI, Brown KR and Cherniack RM: Predicting survival in idiopathic pulmonary fibrosis: Scoring system and survival model. Am J Respir Crit Care Med. 164:1171–1181. 2001. View Article : Google Scholar : PubMed/NCBI
2	Dempsey OJ: Clinical review: Idiopathic pulmonary fibrosis-past, present and future. Respir Med. 100:1871–1885. 2006. View Article : Google Scholar : PubMed/NCBI
3	Noble PW, Barkauskas CE and Jiang D: Pulmonary fibrosis: Patterns and perpetrators. J Clin Invest. 122:2756–2762. 2012. View Article : Google Scholar : PubMed/NCBI
4	du Bois RM: Strategies for treating idiopathic pulmonary fibrosis. Nat Rev Drug Discov. 9:129–140. 2010. View Article : Google Scholar : PubMed/NCBI
5	Wynn TA: Cellular and molecular mechanisms of fibrosis. J Pathol. 214:199–210. 2008. View Article : Google Scholar
6	Wynn TA: Integrating mechanisms of pulmonary fibrosis. J Exp Med. 208:1339–1350. 2011. View Article : Google Scholar : PubMed/NCBI
7	Todd NW, Luzina IG and Atamas SP: Molecular and cellular mechanisms of pulmonary fibrosis. Fibrogenesis Tissue Repair. 5:112012. View Article : Google Scholar : PubMed/NCBI
8	Phan SH: Biology of fibroblasts and myofibroblasts. Proc Am Thorac Soc. 5:334–337. 2008. View Article : Google Scholar : PubMed/NCBI
9	Cutroneo KR, White SL, Phan SH and Ehrlich HP: Therapies for bleomycin induced lung fibrosis through regulation of TGF-beta1 induced collagen gene expression. J Cell Physiol. 211:585–589. 2007. View Article : Google Scholar : PubMed/NCBI
10	Sivakumar P, Ntolios P, Jenkins G and Laurent G: Into the matrix: Targeting fibroblasts in pulmonary fibrosis. Curr Opin Pulm Med. 18:462–469. 2012. View Article : Google Scholar : PubMed/NCBI
11	Lee CG, Cho S, Homer RJ and Elias JA: Genetic control of transforming growth factor-beta1-induced emphysema and fibrosis in the murine lung. Proc Am Thorac Soc. 3:476–477. 2006. View Article : Google Scholar : PubMed/NCBI
12	Caraci F, Gili E, Calafiore M, Failla M, La Rosa C, Crimi N, Sortino MA, Nicoletti F, Copani A and Vancheri C: TGF-beta1 targets the GSK-3beta/beta-catenin pathway via ERK activation in the transition of human lung fibroblasts into myofibroblasts. Pharmacol Res. 57:274–282. 2008. View Article : Google Scholar : PubMed/NCBI
13	Wynn TA and Ramalingam TR: Mechanisms of fibrosis: Therapeutic translation for fibrotic disease. Nat Med. 18:1028–1040. 2012. View Article : Google Scholar : PubMed/NCBI
14	Pourgholamhossein F, Rasooli R, Pournamdari M, Pourgholi L, Samareh-Fekri M, Ghazi-Khansari M, Iranpour M, Poursalehi HR, Heidari MR and Mandegary A: Pirfenidone protects against paraquat-induced lung injury and fibrosis in mice by modulation of inflammation, oxidative stress, and gene expression. Food Chem Toxicol. 112:39–46. 2018. View Article : Google Scholar
15	Romano M, Cianci E, Simiele F and Recchiuti A: Lipoxins and aspirin-triggered lipoxins in resolution of inflammation. Eur J Pharmacol. 760:49–63. 2015. View Article : Google Scholar : PubMed/NCBI
16	Xia J, Zhou XL, Zhao Y, Zhu YQ, Jiang S and Ni SZ: Roles of lipoxin A4 in preventing paracetamol-induced acute hepatic injury in a rabbit model. Inflammation. 36:1431–1439. 2013. View Article : Google Scholar : PubMed/NCBI
17	Jiang X, Li Z, Jiang S, Tong X, Zou X, Wang W, Zhang Z, Wu L and Tian D: Lipoxin A4 exerts protective effects against experimental acute liver failure by inhibiting the NF-κB pathway. Int J Mol Med. 37:773–780. 2016. View Article : Google Scholar : PubMed/NCBI
18	Börgeson E, Docherty NG, Murphy M, Rodgers K, Ryan A, O’Sullivan TP, Guiry PJ, Goldschmeding R, Higgins DF and Godson C: Lipoxin A₄ and benzolipoxin A₄ attenuate experimental renal fibrosis. FASEB J. 25:2967–2979. 2011. View Article : Google Scholar
19	Krönke G, Reich N, Scholtysek C, Akhmetshina A, Uderhardt S, Zerr P, Palumbo K, Lang V, Dees C, Distler O, et al: The 12/15-lipoxygenase pathway counteracts fibroblast activation and experimental fibrosis. Ann Rheum Dis. 71:1081–1087. 2012. View Article : Google Scholar : PubMed/NCBI
20	Martins V, Valença SS, Farias-Filho FA, Molinaro R, Simões RL, Ferreira TPE, Silva PM, Hogaboam CM, Kunkel SL, Fierro IM, et al: ATLa, an aspirin-triggered lipoxin A4 synthetic analog, prevents the inflammatory and fibrotic effects of bleomycin-induced pulmonary fibrosis. J Immunol. 182:5374–5381. 2009. View Article : Google Scholar : PubMed/NCBI
21	Gong J, Guo S, Li HB, Yuan SY, Shang Y and Yao SL: BML-111, a lipoxin receptor agonist, protects haemorrhagic shock-induced acute lung injury in rats. Resuscitation. 83:907–912. 2012. View Article : Google Scholar : PubMed/NCBI
22	Fiore S, Maddox JF, Perez HD and Serhan CN: Identification of a human cDNA encoding a functional high affinity lipoxin A4 receptor. J Exp Med. 180:253–260. 1994. View Article : Google Scholar : PubMed/NCBI
23	Takano T, Fiore S, Maddox JF, Brady HR, Petasis NA and Serhan CN: Aspirin-triggered 15-epilipoxin A4 (LXA4) and LXA4 stable analogues are potent inhibitors of acute inflammation: Evidence for anti-inflammatory receptors. J Exp Med. 185:1693–1704. 1997. View Article : Google Scholar : PubMed/NCBI
24	Gao JL, Chen H, Filie JD, Kozak CA and Murphy PM: Differential expansion of the N-formylpeptide receptor gene cluster in human and mouse. Genomics. 51:270–276. 1998. View Article : Google Scholar : PubMed/NCBI
25	Wang YP, Wu Y, Li LY, Zheng J, Liu RG, Zhou JP, Yuan SY, Shang Y and Yao SL: Aspirin-triggered lipoxin A4 attenuates LPS-induced pro-inflammatory responses by inhibiting activation of NF-κB and MAPKs in BV-2 microglial cells. J Neuroinflammation. 8:952011. View Article : Google Scholar
26	Lee TH, Lympany P, Crea AE and Spur BW: Inhibition of leukotriene B4-induced neutrophil migration by lipoxin A4: structure-function relationships. Biochem Biophys Res Commun. 180:1416–1421. 1991. View Article : Google Scholar : PubMed/NCBI
27	Li HB, Wang GZ, Gong J, Wu ZY, Guo S, Li B, Liu M, Ji YD, Tang M, Yuan SY, et al: BML-111 attenuates hemorrhagic shock-induced acute lung injury through inhibiting activation of mitogen-activated protein kinase pathway in rats. J Surg Res. 183:710–719. 2013. View Article : Google Scholar : PubMed/NCBI
28	Gong J, Li HB, Guo S, Shang Y and Yao SL: Lipoxin receptor agonist, may be a potential treatment for hemorrhagic shock-induced acute lung injury. Med Hypotheses. 79:92–94. 2012. View Article : Google Scholar : PubMed/NCBI
29	Li H, Wu Z, Feng D, Gong J, Yao C, Wang Y, Yuan S, Yao S and Shang Y: BML-111, a lipoxin receptor agonist, attenuates ventilator-induced lung injury in rats. Shock. 41:311–316. 2014. View Article : Google Scholar
30	Zhou XY, Yu ZJ, Yan D, Wang HM, Huang YH, Sha J, Xu FY, Cai ZY and Min WP: BML-11, a lipoxin receptor agonist, protected carbon tetrachloride-induced hepatic fibrosis in rats. Inflammation. 36:1101–1106. 2013. View Article : Google Scholar : PubMed/NCBI
31	Izumo T, Kondo M and Nagai A: Effects of a leukotriene B4 receptor antagonist on bleomycin-induced pulmonary fibrosis. Eur Respir J. 34:1444–1451. 2009. View Article : Google Scholar : PubMed/NCBI
32	Kelly MN, Zheng M, Ruan S, Kolls J, D’Souza A and Shellito JE: Memory CD4⁺ T cells are required for optimal NK cell effector functions against the opportunistic fungal pathogen Pneumocystis murina. J Immunol. 190:285–295. 2013. View Article : Google Scholar
33	Yang J, Nan C, Ripps H and Shen W: Destructive changes in the neuronal structure of the FVB/N mouse retina. PLoS One. 10:e1297192015.
34	AVMA Guodelines for the Euthanasia of Animals. 2013.48
35	Ashcroft T, Simpson JM and Timbrell V: Simple method of estimating severity of pulmonary fibrosis on a numerical scale. J Clin Pathol. 41:467–470. 1988. View Article : Google Scholar : PubMed/NCBI
36	Liu J, Wei Y, Luo Q, Xu F, Zhao Z, Zhang H, Lu L, Sun J, Liu F, Du X, et al: Baicalin attenuates inflammation in mice with OVA-induced asthma by inhibiting NF-κB and suppressing CCR7/CCL19/CCL21. Int J Mol Med. 38:1541–1548. 2016. View Article : Google Scholar : PubMed/NCBI
37	Tang M, Chen L, Li B, Wang Y, Li S, Wen A, Yao S and Shang Y: BML-111 attenuates acute lung injury in endotoxemic mice. J Surg Res. 200:619–630. 2016. View Article : Google Scholar
38	Roach KM, Feghali-Bostwick CA, Amrani Y and Bradding P: Lipoxin A4 Attenuates constitutive and TGF-β1-dependent profibrotic activity in human lung myofibroblasts. J Immunol. 195:2852–2860. 2015. View Article : Google Scholar : PubMed/NCBI
39	Wu SH, Wu XH, Lu C, Dong L and Chen ZQ: Lipoxin A4 inhibits proliferation of human lung fibroblasts induced by connective tissue growth factor. Am J Respir Cell Mol Biol. 34:65–72. 2006. View Article : Google Scholar
40	Mendoza JA, Jacob Y, Cassonnet P and Favre M: Human papillomavirus type 5 E6 oncoprotein represses the transforming growth factor beta signaling pathway by binding to SMAD3. J Virol. 80:12420–12424. 2006. View Article : Google Scholar : PubMed/NCBI
41	Hasegawa M, Matsushita Y, Horikawa M, Higashi K, Tomigahara Y, Kaneko H, Shirasaki F, Fujimoto M, Takehara K and Sato S: A novel inhibitor of Smad-dependent transcriptional activation suppresses tissue fibrosis in mouse models of systemic sclerosis. Arthritis Rheum. 60:3465–3475. 2009. View Article : Google Scholar : PubMed/NCBI
42	Runyan CE, Schnaper HW and Poncelet AC: The phosphatidylinositol 3-kinase/Akt pathway enhances Smad3-stimulated mesangial cell collagen I expression in response to transforming growth factor-beta1. J Biol Chem. 279:2632–2639. 2004. View Article : Google Scholar
43	Xiao L, Du Y, Shen Y, He Y, Zhao H and Li Z: TGF-beta 1 induced fibroblast proliferation is mediated by the FGF-2/ERK pathway. Front Biosci (Landmark Ed). 17:2667–2674. 2012. View Article : Google Scholar
44	Hinz B: Myofibroblasts. Exp Eye Res. 142:56–70. 2016. View Article : Google Scholar
45	Mouratis MA and Aidinis V: Modeling pulmonary fibrosis with bleomycin. Curr Opin Pulm Med. 17:355–361. 2011. View Article : Google Scholar : PubMed/NCBI
46	Kolb M, Margetts PJ, Anthony DC, Pitossi F and Gauldie J: Transient expression of IL-1beta induces acute lung injury and chronic repair leading to pulmonary fibrosis. J Clin Invest. 107:1529–1536. 2001. View Article : Google Scholar : PubMed/NCBI
47	Sime PJ, Marr RA, Gauldie D, Xing Z, Hewlett BR, Graham FL and Gauldie J: Transfer of tumor necrosis factor-alpha to rat lung induces severe pulmonary inflammation and patchy interstitial fibrogenesis with induction of transforming growth factor-beta1 and myofibroblasts. Am J Pathol. 153:825–832. 1998. View Article : Google Scholar : PubMed/NCBI
48	Sheppard D: Transforming growth factor beta: A central modulator of pulmonary and airway inflammation and fibrosis. Proc Am Thorac Soc. 3:413–417. 2006. View Article : Google Scholar : PubMed/NCBI