Monoclonal antibody against Toll-like receptor 4 attenuates ventilator-induced lung injury in rats by inhibiting MyD88- and NF-κB-dependent signaling

Huang,Cuiyuan; Pan,Linghui; Lin,Fei; Dai,Huijun; Fu,Ruili

doi:10.3892/ijmm.2017.2873

Monoclonal antibody against Toll-like receptor 4 attenuates ventilator-induced lung injury in rats by inhibiting MyD88- and NF-κB-dependent signaling

Authors:
- Cuiyuan Huang
- Linghui Pan
- Fei Lin
- Huijun Dai
- Ruili Fu
View Affiliations

Affiliations: Department of Anesthesiology, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: February 1, 2017 https://doi.org/10.3892/ijmm.2017.2873
Pages: 693-700

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

The mechanisms through which mechanical ventilation causes non-infectious inflammatory diseases and lung injury are poorly understood. Animals models of this type of injury suggest that it involves signaling mediated by Toll‑like receptor (TLR)4 and 9. In this study, in order to gain further insight into the involvement of TLR4 in this type of injury, we performed in vivo and in vitro experiments to determine the mechanisms through which TLR4 triggers inflammation. We also examined whether the use of TLR4 monoclonal antibody (mAb) can alleviate this type of injury. For this purpose, rats were tracheotomized and administered intratracheal injections of anti‑TLR4 mAb or saline, and then ventilated for 4 h at a high tidal volume (HTV) of 40 ml/ kg or allowed to breathe spontaneously for the same period of time (controls). Alveolar macrophages (AMs) were isolated from the bronchoalveolar lavage fluid (BALF) of the rats and stimulated for 16 h with tumor necrosis factor (TNF)‑α in the presence or absence of anti‑TLR4 mAb. Lung injury was assessed by examining lung histopathology, lung wet/dry weight ratio, BALF total protein and cytokine levels in BALF and plasma. The mRNA and protein expression levels of TLR4, TLR9, myeloid differentiation factor 88 (Myd88) and nuclear factor (NF)‑κB were measured in cultured macrophages. Compared to the controls (spontaneous breathing), the ventilated rats exhibited greater pulmonary permeability, more severe inflammatory cell infiltration/lung edema, and higher levels of interleukin (IL)‑1β, IL‑6 and TNF‑α in BALF and plasma. The AMs from the ventilated rats expressed higher mRNA and protein levels of TLR4, TLR9, Myd88 and NF‑κB compared with the macrophages from the spontaneously breathing rats. The ventilated rats pre‑treated with anti‑TLR4 mAb exhibited markedly attenuated signs of ventilation‑induced injury, such as less lung inflammation and pulmonary edema, fewer cells in BALF, and lower levels of ILs and TNF‑α in BALF and plasma. Similarly, the TNF‑α‑dependent increases in the mRNA and protein expression of TLR4, Myd88 and NF‑κB in AMs were attenuated when TNF‑α was co‑administered with anti‑TLR4 mAb than when TNF-α was administered alone. Co‑administering anti-TLR4 mAb also reduced the TNF‑α‑dependent secretion of ILs. On the whole, our data demonstrate that TLR4 contributes significantly to ventilation‑induced lung injury by activating the Myd88/NF‑κB pathway, and pre‑treating rats with anti‑TLR4 mAb partially protects them against this type of injury by inhibiting Myd88/NF-κB signaling.

View Figures

View References

1	The Acute Respiratory Distress Syndrome Network: Ventilation with lower tidal volumes as compared with traditional tidal volumes for acute lung injury and the acute respiratory distress syndrome. N Engl J Med. 342:1301–1308. 2000. View Article : Google Scholar
2	Parker JC, Hernandez LA and Peevy KJ: Mechanisms of ventilator-induced lung injury. Crit Care Med. 21:131–143. 1993. View Article : Google Scholar : PubMed/NCBI
3	Frank JA, Wray CM, McAuley DF, Schwendener R and Matthay MA: Alveolar macrophages contribute to alveolar barrier dysfunction in ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 291:L1191–L1198. 2006. View Article : Google Scholar : PubMed/NCBI
4	Kawauchi Y, Takagi H, Hanafusa K, Kono M, Yamatani M and Kojima N: SIGNR1-mediated phagocytosis, but not SIGNR1-mediated endocytosis or cell adhesion, suppresses LPS-induced secretion of IL-6 from murine macrophages. Cytokine. 71:45–53. 2015. View Article : Google Scholar
5	Folco EJ, Sukhova GK, Quillard T and Libby P: Moderate hypoxia potentiates interleukin-1β production in activated human macrophages. Circ Res. 115:875–883. 2014. View Article : Google Scholar : PubMed/NCBI
6	Gordon S: Pattern recognition receptors: doubling up for the innate immune response. Cell. 111:927–930. 2002. View Article : Google Scholar
7	Slutsky AS and Ranieri VM: Ventilator-induced lung injury. N Engl J Med. 369:2126–2136. 2013. View Article : Google Scholar : PubMed/NCBI
8	Zhu S, Pan L, Lin F, Zhao Q and Wei Y: Expression of Toll-like receptor 3 and 4 in the lung tissue of rats with ventilator-induced lung injury. J Clin Anesthesiol. 29:594–597. 2013.
9	Dai H, Pan L, Lin F, Ge W, Li W and He S: Role and mechanism of signal pathway mediated by Toll-like receptor 9-myeloid differentiation factor 88 in alveolar macrophages in ventilator-induced lung injury in rats. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 26:289–293. 2014.In Chinese. PubMed/NCBI
10	Liu Y, Yin H, Zhao M and Lu Q: TLR2 and TLR4 in autoimmune diseases: a comprehensive review. Clin Rev Allergy Immunol. 47:136–147. 2013. View Article : Google Scholar : PubMed/NCBI
11	Starkhammar M, Larsson O, Kumlien Georén S, Leino M, Dahlén SE, Adner M and Cardell LO: Toll-like receptor ligands LPS and poly (I:C) exacerbate airway hyperresponsiveness in a model of airway allergy in mice, independently of inflammation. PLoS One. 9:e1041142014. View Article : Google Scholar : PubMed/NCBI
12	Pfeffer K, Matsuyama T, Kündig TM, Wakeham A, Kishihara K, Shahinian A, Wiegmann K, Ohashi PS, Krönke M and Mak TW: Mice deficient for the 55 kd tumor necrosis factor receptor are resistant to endotoxic shock, yet succumb to L. monocytogenes infection. Cell. 73:457–467. 1993. View Article : Google Scholar : PubMed/NCBI
13	Bertok S, Wilson MR, Morley PJ, de Wildt R, Bayliffe A and Takata M: Selective inhibition of intra-alveolar p55 TNF receptor attenuates ventilator-induced lung injury. Thorax. 67:244–251. 2012. View Article : Google Scholar :
14	Li C, Li B, Dong Z, Gao L, He X, Liao L, Hu C, Wang Q and Jin Y: Lipopolysaccharide differentially affects the osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells through Toll-like receptor 4 mediated nuclear factor κB pathway. Stem Cell Res Ther. 5:672014. View Article : Google Scholar
15	Geissmann F, Manz MG, Jung S, Sieweke MH, Merad M and Ley K: Development of monocytes, macrophages, and dendritic cells. Science. 327:656–661. 2010. View Article : Google Scholar : PubMed/NCBI
16	Müller HC, Hellwig K, Rosseau S, Tschernig T, Schmiedl A, Gutbier B, Schmeck B, Hippenstiel S, Peters H, Morawietz L, et al: Simvastatin attenuates ventilator-induced lung injury in mice. Crit Care. 14:R1432010. View Article : Google Scholar : PubMed/NCBI
17	Ko YA, Yang MC, Huang HT, Hsu CM and Chen LW: NF-κB activation in myeloid cells mediates ventilator-induced lung injury. Respir Res. 14:692013. View Article : Google Scholar
18	Mortaz E, Adcock IM, Ito K, Kraneveld AD, Nijkamp FP and Folkerts G: Cigarette smoke induces CXCL8 production by human neutrophils via activation of TLR9 receptor. Eur Respir J. 36:1143–1154. 2010. View Article : Google Scholar
19	Haitsma JJ, Uhlig S, Göggel R, Verbrugge SJ, Lachmann U and Lachmann B: Ventilator-induced lung injury leads to loss of alveolar and systemic compartmentalization of tumor necrosis factor-alpha. Intensive Care Med. 26:1515–1522. 2000. View Article : Google Scholar : PubMed/NCBI
20	Mukherjee S and Biswas T: Activation of TOLLIP by porin prevents TLR2-associated IFN-γ and TNF-α-induced apoptosis of intestinal epithelial cells. Cell Signal. 26:2674–2682. 2014. View Article : Google Scholar : PubMed/NCBI
21	Anthwal A, Thakur BK, Rawat MS, Rawat DS, Tyagi AK and Aggarwal BB: Synthesis, characterization and in vitro anticancer activity of C-5 curcumin analogues with potential to inhibit TNF-α-induced NF-κB activation. Biomed Res Int. 2014:5241612014. View Article : Google Scholar
22	Fioretto JR and Carvalho WB: Temporal evolution of acute respiratory distress syndrome definitions. J Pediatr (Rio J). 89:523–530. 2013. View Article : Google Scholar
23	Monahan LJ: Acute respiratory distress syndrome. Curr Probl Pediatr Adolesc Health Care. 43:278–284. 2013. View Article : Google Scholar : PubMed/NCBI
24	Pan WZ, Shi CX, Tian M and Yu JG: Anti-CD11c antibody, Efalizumab attenuate ventilator-induced lung injury. Eur Rev Med Pharmacol Sci. 18:2182–2190. 2014.PubMed/NCBI
25	Akella A, Sharma P, Pandey R and Deshpande SB: Characterization of oleic acid-induced acute respiratory distress syndrome model in rat. Indian J Exp Biol. 52:712–719. 2014.PubMed/NCBI
26	Belperio JA, Keane MP, Lynch JP III and Strieter RM: The role of cytokines during the pathogenesis of ventilator-associated and ventilator-induced lung injury. Semin Respir Crit Care Med. 27:350–364. 2006. View Article : Google Scholar : PubMed/NCBI
27	Meylan E, Tschopp J and Karin M: Intracellular pattern recognition receptors in the host response. Nature. 442:39–44. 2006. View Article : Google Scholar : PubMed/NCBI
28	O'Neill LA and Bowie AG: The family of five: TIR-domain-containing adaptors in Toll-like receptor signalling. Nat Rev Immunol. 7:353–364. 2007. View Article : Google Scholar : PubMed/NCBI
29	He Z, Gao Y, Deng Y, Li W, Chen Y, Xing S, Zhao X, Ding J and Wang X: Lipopolysaccharide induces lung fibroblast proliferation through Toll-like receptor 4 signaling and the phosphoinositide3-kinase-Akt pathway. PLoS One. 7:e359262012. View Article : Google Scholar : PubMed/NCBI
30	Prakash A, Mesa KR, Wilhelmsen K, Xu F, Dodd-o JM and Hellman J: Alveolar macrophages and Toll-like receptor 4 mediate ventilated lung ischemia reperfusion injury in mice. Anesthesiology. 117:822–835. 2012. View Article : Google Scholar : PubMed/NCBI
31	Perkins DJ and Vogel SN: Inflammation: Species-specific TLR signalling - insight into human disease. Nat Rev Rheumatol. 12:198–200. 2016. View Article : Google Scholar : PubMed/NCBI
32	Hayes M, Curley GF, Masterson C, Contreras M, Ansari B, Devaney J, O'Toole D and Laffey JG: Pulmonary overexpression of inhibitor κBα decreases the severity of ventilator-induced lung injury in a rat model. Br J Anaesth. 113:1046–1054. 2014. View Article : Google Scholar : PubMed/NCBI
33	Bryant CE, Symmons M and Gay NJ: Toll-like receptor signalling through macromolecular protein complexes. Mol Immunol. 63:162–165. 2014. View Article : Google Scholar : PubMed/NCBI
34	Hazen SL: Oxidized phospholipids as endogenous pattern recognition ligands in innate immunity. J Biol Chem. 283:15527–15531. 2008. View Article : Google Scholar : PubMed/NCBI
35	Jiang D, Liang J, Fan J, Yu S, Chen S, Luo Y, Prestwich GD, Mascarenhas MM, Garg HG, Quinn DA, et al: Regulation of lung injury and repair by Toll-like receptors and hyaluronan. Nat Med. 11:1173–1179. 2005. View Article : Google Scholar : PubMed/NCBI
36	Oya K, Sakamoto N and Sato M: Hypoxia suppresses stretch-induced elongation and orientation of macrophages. Biomed Mater Eng. 23:463–471. 2013.PubMed/NCBI
37	Wu J, Yan Z, Schwartz DE, Yu J, Malik AB and Hu G: Activation of NLRP3 inflammasome in alveolar macrophages contributes to mechanical stretch-induced lung inflammation and injury. J Immunol. 190:3590–3599. 2013. View Article : Google Scholar : PubMed/NCBI
38	Dai H, Pan L, Lin F, Ge W, Li W and He S: Mechanical ventilation modulates Toll-like receptors 2, 4, and 9 on alveolar macrophages in a ventilator-induced lung injury model. J Thorac Dis. 7:616–624. 2015.PubMed/NCBI
39	Wu GJ, Chen TL, Ueng YF and Chen RM: Ketamine inhibits tumor necrosis factor-alpha and interleukin-6 gene expressions in lipopolysaccharide-stimulated macrophages through suppression of toll-like receptor 4-mediated c-Jun N-terminal kinase phosphorylation and activator protein-1 activation. Toxicol Appl Pharmacol. 228:105–113. 2008. View Article : Google Scholar : PubMed/NCBI
40	Yang P, Yang N, Zhang X and Xu X: The significance and mechanism of propofol on treatment of ischemia reperfusion induced lung injury in rats. Cell Biochem Biophys. 70:1527–1532. 2014. View Article : Google Scholar : PubMed/NCBI
41	Spieth PM, Carvalho AR, Güldner A, Pelosi P, Kirichuk O, Koch T and de Abreu MG: Effects of different levels of pressure support variability in experimental lung injury. Anesthesiology. 110:342–350. 2009.PubMed/NCBI
42	Biehl M, Kashiouris MG and Gajic O: Ventilator-induced lung injury: Minimizing its impact in patients with or at risk for ARDS. Respir Care. 58:927–937. 2013. View Article : Google Scholar : PubMed/NCBI