Aspirin modulates the inflammatory response in a thrombus‑stimulated LMVEC model

Wang,Lingcong; Ying,Rongbiao; Jiang,Huifang; Jin,Qun; Kuang,Jing; Zhang,Zhirong; Shi,Ying; Cai,Danli; Yang,Ruhui

doi:10.3892/ijmm.2018.3561

Aspirin modulates the inflammatory response in a thrombus‑stimulated LMVEC model

Authors:
- Lingcong Wang
- Rongbiao Ying
- Huifang Jiang
- Qun Jin
- Jing Kuang
- Zhirong Zhang
- Ying Shi
- Danli Cai
- Ruhui Yang
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Affiliations: Department of ICU, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310006, P.R. China, Department of Surgical Oncology, Tumor Hospital of Taizhou, Wenling, Zhejiang 317502, P.R. China, Department of Hematology, Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang 310012, P.R. China, Department of Pneumology, Zhejiang University International Hospital, Shulan (Hangzhou) Hospital, Hangzhou, Zhejiang 310000, P.R. China, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China, Lishui University, Lishui, Zhejiang 323000, P.R. China
Published online on: March 13, 2018 https://doi.org/10.3892/ijmm.2018.3561
Pages: 3253-3266
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The purpose of the present study was to examine whether aspirin interferes with the inflammatory response in a thrombus‑stimulated lung microvascular endothelial cell (LMVEC) model. The LMVECs were randomly divided into eight groups: Normal group (group N), model group (group M), model + ASP group (group M+A), model+CX3CL1‑short hairpin (sh)RNA group (group M+SH), model + CX3CL1‑overexpression vector group (group M+CX3), model + ASP + shRNA group (group M+A+SH), model + ASP + CX3CL1‑overexpression vector group (group M+A+CX3), and normal + virus control group (group N+V). The endothelial cells were cultured, and a thrombus was added to the cells. Briefly, 12 h following the precipitation of the thrombus, data from ELISA, reverse transcription‑quantitative polymerase chain reaction analysis and confocal microscopy revealed that the levels of tumor necrosis factor (TNF)‑α, interleukin (IL)‑6, CX3C chemokine ligand 1 (CX3CL1), CX3C chemokine receptor 1 (CX3CR1) and nuclear factor‑κB (NF‑κB) in group M were increased, compared with those in group N (P<0.01). These levels, with the exception of TNF‑α, were significantly lower in group M+SH, compared with those in group M (P<0.01). Furthermore, the levels of IL‑6 in groups M+A, M+CX3 and M+A+CX3 were decreased, compared with those in group M (P<0.01); the level of TNF‑α in group M+A+SH was decreased, compared with that in group M (P<0.01); the level of CX3CR1 waslower in groups M+A and M+A+SH, compared with that in group M (P<0.01), and the level of NF‑κB in group M+SH was decreased, compared with the level in group M and group M+A (P<0.05). In conclusion, the thrombus‑stimulated LMVEC model exhibited induced production of TNF‑α, IL‑6, CX3CL, CX3CR1, NF‑κB and intercellular adhesion molecule‑1. Furthermore, it was confirmed that the signaling pathways involving CX3CL1‑NF‑κB, IL‑6 and TNF‑α were partly inhibited by aspirin.

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1	Saghazadeh A, Hafizi S and Rezaei N: Inflammation in venous thromboembolism: Cause or consequence? Int Immunopharmacol. 28:655–665. 2015. View Article : Google Scholar : PubMed/NCBI
2	Deng C, Zhai Z, Wu D, Lin Q, Yang Y, Yang M, Ding H, Cao X, Zhang Q and Wang C: Inflammatory response and pneumocyte apoptosis during lung ischemia-reperfusion injury in an experimental pulmonary thromboembolism model. J Thromb Thrombolysis. 40:42–53. 2015. View Article : Google Scholar : PubMed/NCBI
3	Wang LC, Wu JN, Xia GL, Mao W, Ying RB, Huang LQ and Jiang GL: Effect of aspirin on fractalkine in rats with pulmonary embolism. Trop J Pharm Res. 13:753–760. 2014. View Article : Google Scholar
4	Wang LC, Jiang RL, Zhang W, Wei LL and Yang RH: Effects of aspirin on the expression of nuclear factor-κB in a rat model of acute pulmonary embolism. World J Emerg Med. 5:229–233. 2014. View Article : Google Scholar
5	Jiang R, Wei L, Zhu M, Wu J and Wang L: Aspirin inhibits LPS-induced expression of PI3K/Akt, ERK, NF-κB, CX3CL1, and MMPs in human bronchial epithelial cells. Inflammation. 39:643–650. 2016. View Article : Google Scholar
6	Felding-Habermann B, Habermann R, Saldívar E and Ruggeri ZM: Role of beta3 integrins in melanoma cell adhesion to activated platelets under flow. J Biol Chem. 271:5892–5900. 1996. View Article : Google Scholar : PubMed/NCBI
7	Terrisse AD, Puech N, Allart S, Gourdy P, Xuereb JM, Payrastre B and Sié P: Internalization of microparticles by endothelial cells promotes platelet/endothelial cell interaction under flow. J Thromb Haemost. 8:2810–2819. 2010. View Article : Google Scholar : PubMed/NCBI
8	Liu X, Liu J, Zhao S, Zhang H, Cai W, Cai M, Ji X, Leak RK, Gao Y, Chen J and Hu X: Interleukin-4 is essential for Microglia/Macrophage M2 polarization and long-term recovery after cerebral ischemia. Stroke. 47:498–504. 2016. View Article : Google Scholar : PubMed/NCBI
9	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
10	Malenovska H: Virus quantitation by transmission electron microscopy, TCID₅₀, and the role of timing virus harvesting: A case study of three animal viruses. J Virol Methods. 191:136–140. 2013. View Article : Google Scholar : PubMed/NCBI
11	Bazan JF, Bacon KB, Hardiman G, Wang W, Soo K, Rossi D, Greaves DR, Zlotnik A and Schall TJ: A new class of membrane-bound chemokine with a CX3C motif. Nature. 385:640–644. 1997. View Article : Google Scholar : PubMed/NCBI
12	Todorova D, Sabatier F, Doria E, Lyonnet L, Vacher Coponat H, Robert S, Despoix N, Legris T, Moal V, Loundou A, et al: Fractalkine expression induces endothelial progenitor cell lysis by natural killer cells. PLoS One. 6:e266632011. View Article : Google Scholar : PubMed/NCBI
13	Matsumiya T, Ota K, Imaizumi T, Yoshida H, Kimura H and Satoh K: Characterization of synergistic induction of CX3CL1/fractalkine by TNF-alpha and IFN-gamma in vascular endothelial cells: An essential role for TNF-alpha in post-transcriptional regulation of CX3CL1. J Immunol. 184:4205–4214. 2010. View Article : Google Scholar : PubMed/NCBI
14	Szukiewicz D, Kochanowski J, Mittal TK, Pyzlak M, Szewczyk G and Cendrowski K: CX3CL1 (fractalkine) and TNFα production by perfused human placental lobules under normoxic and hypoxic conditions in vitro: The importance of CX3CR1 signaling. Inflamm Res. 63:179–189. 2014. View Article : Google Scholar
15	Cardona AE, Pioro EP, Sasse ME, Kostenko V, Cardona SM, Dijkstra IM, Huang D, Kidd G, Dombrowski S, Dutta R, et al: Control of microglial neurotoxicity by the fractalkine receptor. Nat Neurosci. 9:917–924. 2006. View Article : Google Scholar : PubMed/NCBI
16	Lyons A, Lynch AM, Downer EJ, Hanley R, O'Sullivan JB, Smith A and Lynch MA: Fractalkine-induced activation of the phosphatidylinositol-3 kinase pathway attentuates microglial activation in vivo and in vitro. J Neurochem. 110:1547–1556. 2009. View Article : Google Scholar : PubMed/NCBI
17	Sukkar MB, Issa R, Xie S, Oltmanns U, Newton R and Chung KF: Fractalkine/CX3CL1 production by human airway smooth muscle cells: Induction by IFN-gamma and TNF-alpha and regulation by TGF-beta and corticosteroids. Am J Physiol Lung Cell Mol Physiol. 287:L1230–L1240. 2004. View Article : Google Scholar : PubMed/NCBI
18	Li F, Zhang H, Xu KY, Wei Q and Zhou GX: Role of the chemokine fractalkine in a rat model of acute necrotizing pancreatitis and the interventional effect of ulinastatin. Arch Iran Med. 16:83–87. 2013.PubMed/NCBI
19	Briones TL, Woods J and Wadowska M: Chronic neuroinflammation and cognitive impairment following transient global cerebral ischemia: Role of fractalkine/CX3CR1 signaling. J Neuroinflammation. 11:132014. View Article : Google Scholar : PubMed/NCBI
20	Tsang JY, Ni YB, Chan SK, Shao MM, Kwok YK, Chan KW, Tan PH and Tse GM: CX3CL1 expression is associated with poor outcome in breast cancer patients. Breast Cancer Res Treat. 140:495–504. 2013. View Article : Google Scholar : PubMed/NCBI
21	Volin MV, Huynh N, Klosowska K, Reyes RD and Woods JM: Fractalkine-induced endothelial cell migration requires MAP kinase signaling. Pathobiology. 77:7–16. 2010. View Article : Google Scholar : PubMed/NCBI
22	Ars C, Thurion P, Delos M, Sibille Y and Pilette C: Small airway obstruction in severe pulmonary arterial hypertension correlates with increased airway CD8⁺ T-cells and fractalkine expression. Eur Respir J. 34:1494–1496. 2009. View Article : Google Scholar : PubMed/NCBI
23	Yang RC, Chang CC, Sheen JM, Wu HT, Pang JH and Huang ST: Davallia bilabiata inhibits TNF-α-induced adhesion molecules and chemokines by suppressing IKK/NF-kappa B pathway in vascular endothelial cells. Am J Chin Med. 42:1411–1429. 2014. View Article : Google Scholar
24	Cimato TR and Palka BA: Fractalkine (CX3CL1), GM-CSF and VEGF-a levels are reduced by statins in adult patients. Clin Transl Med. 3:142014. View Article : Google Scholar : PubMed/NCBI
25	Cao N, Chen T, Guo ZP, Qin S and Li MM: Monoammonium glycyrrhizate suppresses tumor necrosis factor-α induced chemokine production in HMEC-1 cells, possibly by blocking the translocation of nuclear factor-kB into the nucleus. Can J Physiol Pharmacol. 92:859–865. 2014. View Article : Google Scholar : PubMed/NCBI
26	Guo X, Pan Y, Xiao C, Wu Y, Cai D and Gu J: Fractalkine stimulates cell growth and increases its expression via NF-κB pathway in RA-FLS. Int J Rheum Dis. 15:322–329. 2012. View Article : Google Scholar : PubMed/NCBI
27	Anbarasan C, Bavanilatha M, Latchumanadhas K and Ajit Mullasari S: ICAM-1 molecular mechanism and genome wide SNP's association studies. Indian Heart J. 67:282–287. 2015. View Article : Google Scholar : PubMed/NCBI
28	Lee SJ, Drabik K, Van Wagoner NJ, Lee S, Choi C, Dong Y and Benveniste EN: ICAM-1-induced expression of proinflammatory cytokines in astrocytes: Involvement of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways. J Immunol. 165:4658–4666. 2000. View Article : Google Scholar : PubMed/NCBI
29	Liu JF, Tsao YT and Hou CH: Fractalkine/CX3CL1 induced intercellular adhesion molecule-1-dependent tumor metastasis through the CX3CR1/pI3K/Akt/NF-κB pathway in human osteosarcoma. Oncotarget. 8:54136–54148. 2016.
30	Tarantino E, Amadio P, Squellerio I, Porro B, Sandrini L, Turnu L, Cavalca V, Tremoli E and Barbieri SS: Role of thromboxane-dependent platelet activation in venous thrombosis: Aspirin effects in mouse model. Pharmacol Res. 107:415–425. 2016. View Article : Google Scholar : PubMed/NCBI
31	Zhang F, Lu M, Wang H and Ren T: Aspirin attenuates angiotensin II-induced inflammation in bone marrow mesenchymal stem cells via the inhibition of ERK1/2 and NF-κB activation. Biomed Rep. 1:930–934. 2013. View Article : Google Scholar
32	Stark LA, Din FV, Zwacka RM and Dunlop MG: Aspirin-induced activation of the NF-κB signaling pathway: A novel mechanism for aspirin-mediated apoptosis in colon cancer cells. Faseb J. 15:1273–1275. 2001. View Article : Google Scholar : PubMed/NCBI
33	Chang MC, Hung HP, Lin LD, Shyu YC, Wang TM, Lin HJ, Chan CP, Huang CC and Jeng JH: Effect of interleukin-1β on ICAM-1 expression of dental pulp cells: Role of PI3K/Akt, MEK/ERK, and cyclooxygenase. Clin Oral Investig. 19:117–126. 2015. View Article : Google Scholar
34	Szukiewicz D, Wojciechowska M, Bilska A, Stangret A, Szewczyk G, Mittal TK, Watroba M and Kochanowski J: Aspirin action in endothelial cells: Different patterns of response between chemokine CX3CL1/CX3CR1 and TNF-α/TNFR1 signaling pathways. Cardiovasc Drugs Ther. 29:219–229. 2015. View Article : Google Scholar : PubMed/NCBI