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. 2013 Dec 10;9(1):133–140. doi: 10.2478/s11536-013-0245-7

Simvastatin attenuates the lipopolysaccharideinduced inflammatory response of rat pulmonary microvascular endothelial cells by downregulating toll-like receptor 4 expression

Yi Li 1, Jian-yong Ding 2, Biao-xue Ge 3, Chang-hong Miao 4,✉
PMCID: PMC7101868  PMID: 32288932

Abstract

Objective

The therapeutic potential of simvastatin as an anti-inflammatory agent was explored by investigating its effect on the lipopolysaccharide (LPS)-induced inflammatory response in rat pulmonary microvascular endothelial cells (RPMVECs).

Methods

RPMVECs were isolated and the mRNA and protein levels of different toll-like receptors (TLR) were assessed by qRT-PCR and western blotting. The LPS-induced expressions of TLR4, TNF-α and iNOS were analyzed in RPMVECs treated with different concentrations of simvastatin for different times. NF-κB activation was examined by immuofluroscence, luciferase reporter assay and western blotting.

Results

TLR4 is abundantly expressed in RPMVECs, and its expression is induced by LPS stimulation. Simvastatin inhibited LPS-induced TLR4 expression at the mRNA and protein levels in a time-dependent manner (p<0.01), and alleviated inflammation in RPMVECs by inhibiting the release of inflammatory factors such as TNF-α and iNOS. Further study indicated that simvastatin significantly attenuated NF-κB activity by inhibiting the degradation of IκB-α. Pretreatment with pyrrolidine dithiocarbamate (PDTC) and knock-down of TLR4 expression by RNA interference down-regulated the LPS-induced inflammatory response in RPMVECs.

Conclusion

Simvastatin inhibits the LPS-induced inflammatory response in RPMVECs by down-regulating TLR4 expression, suggesting its role as a potential inhibitor of LPS-induced inflammation

Keywords: Simvastatin, Pulmonary microvascular endothelial cells, TLR4, LPS, Mechanism

References

  • [1].Doyle RL, Szaflarski N, Modin GW, Wiener-Kronish JP, Matthay MA. Identification of patients with acute lung injury. Predictors of mortality. Am J Respir Crit Care Med. 1995;152(6Pt1):1818–1824. doi: 10.1164/ajrccm.152.6.8520742. [DOI] [PubMed] [Google Scholar]
  • [2].Wu JS, Sheng L, Wang SH, et al. The impact of clinical risk factors in the conversion from acute lung injury to acute respiratory distress syndrome in severe multiple trauma patients. J Int Med Res. 2008;36(3):579–586. doi: 10.1177/147323000803600325. [DOI] [PubMed] [Google Scholar]
  • [3].Akira S, Takeda K, Kaisho T. Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol. 2001;2(8):675–680. doi: 10.1038/90609. [DOI] [PubMed] [Google Scholar]
  • [4].Branger J, Knapp S, Weijer S, et al. Role of Tolllike receptor 4 in gram-positive and gram-negative pneumonia in mice. Infect Immun. 2004;72(2):788–794. doi: 10.1128/IAI.72.2.788-794.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [5].Basu S, Fenton MJ. Toll-like receptors: function and roles in lung disease. Am J Physiol Lung Cell Mol Physiol. 2004;286(5):L887–892. doi: 10.1152/ajplung.00323.2003. [DOI] [PubMed] [Google Scholar]
  • [6].Pan ZK. Toll-like receptors and TLR-mediated signaling: more questions than answers. Am J Physiol Lung Cell Mol Physiol. 2004;286(5):L918–920. doi: 10.1152/ajplung.00381.2003. [DOI] [PubMed] [Google Scholar]
  • [7].Tobert JA. Lovastatin and beyond: the history of the HMG-CoA reductase inhibitors. Nat Rev Drug Discov. 2003;2(7):517–526. doi: 10.1038/nrd1112. [DOI] [PubMed] [Google Scholar]
  • [8].Stancu C, Sima A. Statins: mechanism of action and effects. J Cell Mol Med. 2001;5(4):378–387. doi: 10.1111/j.1582-4934.2001.tb00172.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [9].Emanuele E, Geroldi D. A novel mechanism of action of atorvastatin against cardiovascular risk: a commentary on “decreased plasma soluble RAGE in patients with hypercholesterolemia: effects of statins”. Free Radic Biol Med. 2007;43(9):1231–1232. doi: 10.1016/j.freeradbiomed.2007.07.008. [DOI] [PubMed] [Google Scholar]
  • [10].Sundararaj KP, Samuvel DJ, Li Y, et al. Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation. J Leukoc Biol. 2008;84(4):1120–1129. doi: 10.1189/jlb.0108064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [11].Rudd JH, Myers KS, Bansilal S, et al. (18) Fluorodeoxyglucose positron emission tomography imaging of atherosclerotic plaque inflammation is highly reproducible: implications for atherosclerosis therapy trials. J Am Coll Cardiol. 2007;50(9):892–896. doi: 10.1016/j.jacc.2007.05.024. [DOI] [PubMed] [Google Scholar]
  • [12].Paoletti R, Gotto AM, Jr., Hajjar DP. Inflammation in atherosclerosis and implications for therapy. Circulation. 2004;109(23Suppl1):III20–26. doi: 10.1161/01.CIR.0000131514.71167.2e. [DOI] [PubMed] [Google Scholar]
  • [13].Ryan US, White LA, Lopez M, Ryan JW. Use of microcarriers to isolate and culture pulmonary microvascular endothelium. Tissue Cell. 1982;14(3):597–606. doi: 10.1016/0040-8166(82)90050-7. [DOI] [PubMed] [Google Scholar]
  • [14].Ke AW, Shi GM, Zhou J, et al. CD151 amplifies signaling by integrin alpha6beta1 to PI3K and induces the epithelial-mesenchymal transition in HCC cells. Gastroenterology. 2011;140(5):1629–1641 e15. doi: 10.1053/j.gastro.2011.02.008. [DOI] [PubMed] [Google Scholar]
  • [15].Mogensen TH. Pathogen recognition and inflammatory signaling in innate immune defenses. Clin Microbiol Rev. 2009;22(2):240–273. doi: 10.1128/CMR.00046-08. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [16].Marshall HE, Potts EN, Kelleher ZT, Stamler JS, Foster WM, Auten RL. Protection from lipopolysaccharide-induced lung injury by augmentation of airway S-nitrosothiols. Am J Respir Crit Care Med. 2009;180(1):11–18. doi: 10.1164/rccm.200807-1186OC. [DOI] [PMC free article] [PubMed] [Google Scholar] [Research Misconduct Found]
  • [17].Andreakos E, Sacre SM, Smith C, et al. Distinct pathways of LPS-induced NF-kappa B activation and cytokine production in human myeloid and nonmyeloid cells defined by selective utilization of MyD88 and Mal/TIRAP. Blood. 2004;103(6):2229–2237. doi: 10.1182/blood-2003-04-1356. [DOI] [PubMed] [Google Scholar]
  • [18].Huber M, Kalis C, Keck S, et al. R-form LPS, the master key to the activation ofTLR4/MD-2-positive cells. Eur J Immunol. 2006;36(3):701–711. doi: 10.1002/eji.200535593. [DOI] [PubMed] [Google Scholar]
  • [19].Sohn EJ, Paape MJ, Connor EE, Bannerman DD, Fetterer RH, Peters RR. Bacterial lipopolysaccharide stimulates bovine neutrophil production of TNF-alpha, IL-1beta, IL-12 and IFN-gamma. Vet Res. 2007;38(6):809–818. doi: 10.1051/vetres:2007033. [DOI] [PubMed] [Google Scholar]
  • [20].Jang CH, Choi JH, Byun MS, Jue DM. Chloroquine inhibits production of TNF-alpha, IL-1beta and IL-6 from lipopolysaccharide-stimulated human monocytes/macrophages by different modes. Rheumatology (Oxford) 2006;45(6):703–710. doi: 10.1093/rheumatology/kei282. [DOI] [PubMed] [Google Scholar]
  • [21].Barnes PJ. Mediators of chronic obstructive pulmonary disease. Pharmacol Rev. 2004;56(4):515–548. doi: 10.1124/pr.56.4.2. [DOI] [PubMed] [Google Scholar]
  • [22].Laza-Stanca V, Stanciu LA, Message SD, Edwards MR, Gern JE, Johnston SL. Rhinovirus replication in human macrophages induces NF-kappaBdependent tumor necrosis factor alpha production. J Virol. 2006;80(16):8248–8258. doi: 10.1128/JVI.00162-06. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [23].Li Y, Dinwiddie DL, Harrod KS, Jiang Y, Kim KC. Anti-inflammatory effect of MUC1 during respiratory syncytial virus infection of lung epithelial cells in vitro. Am J Physiol Lung Cell Mol Physiol. 2010;298(4):L558–563. doi: 10.1152/ajplung.00225.2009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [24].Bereswill S, Munoz M, Fischer A, et al. Antiinflammatory effects of resveratrol, curcumin and simvastatin in acute small intestinal inflammation. PLoS One. 2010;5(12):e15099. doi: 10.1371/journal.pone.0015099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [25].Jahovic N, Gedik N, Ercan F, et al. Effects of statins on experimental colitis in normocholesterolemic rats. Scand J Gastroenterol. 2006;41(8):954–962. doi: 10.1080/00365520600554444. [DOI] [PubMed] [Google Scholar]
  • [26].Hsu HH, Ko WJ, Hsu JY, et al. Simvastatin ameliorates established pulmonary hypertension through a heme oxygenase-1 dependent pathway in rats. Respir Res. 2009;10:32. doi: 10.1186/1465-9921-10-32. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [27].Hothersall E, McSharry C, Thomson NC. Potential therapeutic role for statins in respiratory disease. Thorax. 2006;61(8):729–734. doi: 10.1136/thx.2005.057976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [28].Xiao J, Wang K, Feng YL, Chen XR, Xu D, Zhang MK. Role of extracellular signal-regulated kinase 1/2 in cigarette smoke-induced mucus hypersecretion in a rat model. Chin Med J (Engl) 2011;124(20):3327–3333. [PubMed] [Google Scholar]

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