Cigarette smoke extract stimulates human pulmonary artery smooth muscle cell proliferation: Role of inflammation and oxidative stress

Document Type : Original Article

Authors

Department of Respiratory and Critical Care Medicine, Tianjin Medical University General Hospital, Tianjin 300052, China

Abstract

Objective(s): Cigarette smoke may play a direct role in proliferation of human pulmonary artery smooth muscle cells (HPASMCs). However, the mechanism involved and the effect of interventions remain unclear. We aimed to evaluate the effect of cigarette smoke extract (CSE) on HPASMCs, explore the role of inflammation and oxidative stress, and the effects of Tempol and PDTC in this process.
Materials and Methods: HPASMCs were subjected to normal control (NC), CSE, CSE+Tempol (CSE+T), and CSE+PDTC (CSE+P) groups. Proliferation of HPASMCs was measured by CCK-8 and Western blot. TNF-α, IL-6, MDA, and SOD levels were determined by ELISA and commercial kits. Nuclear translocation of NF-κB p65 was evaluated by western blot.
Results: 1%, 2.5%, and 5% CSE all promoted proliferation of HPASMCs, and effect of 1% CSE was the most significant, however, 7.5% and 10% CSE inhibited viability of cells (all P<0.05). Compared with the NC group, TNF-α, IL-6, and MDA levels increased, SOD activity decreased (all P<0.05), and NF-κB p65 expression in nuclei increased (P=0.04) in the CSE group. Tempol and PDTC inhibited the proliferation of HPASMCs induced by CSE (all P<0.05). And compared with the CSE group, TNF-α, IL-6, and MDA levels in CSE+T and CSE+P groups decreased, while SOD activity increased (all P<0.05). Tempol reduced the expression of NF-κB p65 in nuclei but did not achieve a significant difference (P=0.08). PDTC inhibited the nuclear translocation of NF-κB p65 (P=0.03).
Conclusion: CSE stimulates HPASMCs proliferation in a certain concentration range. The CSE-induced proliferation of HPASMCs involved excessive inflammatory response and oxidative stress. Tempol and PDTC attenuate these effects of CSE on HPASMCs.

Keywords

References

1. Karnati S, Seimetz M, Kleefeldt F, Sonawane A, Madhusudhan T, Bachhuka A, et al. Chronic obstructive pulmonary disease and the cardiovascular system: Vascular repair and regeneration as a therapeutic target. Front Cardiovasc Med 2021; 8: 649512.
2. Gao L, Liu J, Hao Y, Zhao Z, Tan H, Zhang J, et al. Chronic intermittent hypobaric hypoxia attenuates monocrotaline-induced pulmonary arterial hypertension via modulating inflammation and suppressing NF-κB/p38 pathway. Iran J Basic Med Sci 2018; 21: 244-252.
3. Scharf SM, Iqbal M, Keller C, Criner G, Lee S, Fessler HE. Hemodynamic characterization of patients with severe emphysema. Am J Respir Crit Care Med 2002; 166: 314-322.
4. Thabut G, Dauriat G, Stern JB, Logeart D, Lévy A, Marrash-Chahla R, et al. Pulmonary hemodynamics in advanced COPD candidates for lung volume reduction surgery or lung transplantation. Chest 2005; 127: 1531-1536.
5. Nathan SD, Barbera JA, Gaine SP, Harari S, Martinez FJ, Olschewski H, et al. Pulmonary hypertension in chronic lung disease and hypoxia. Eur Respir J 2019; 53: 1801914.
6. Santos S, Peinado VI, Ramírez J, Melgosa T, Roca J, Rodriguez-Roisin R, et al. Characterization of pulmonary vascular remodelling in smokers and patients with mild COPD. Eur Respir J 2002; 19: 632-638.
7. Peinado VI, Pizarro S, Barberà JA. Pulmonary vascular involvement in COPD. Chest 2008; 134: 808-814.
8. Wright JL, Levy RD, Churg A. Pulmonary hypertension in chronic obstructive pulmonary disease: Current theories of pathogenesis and their implications for treatment. Thorax 2005; 60: 605-609.
9. Li Y, Pu G, Chen C, Yang L. Inhibition of FHL1 inhibits cigarette smoke extract-induced proliferation in pulmonary arterial smooth muscle cells. Mol Med Rep 2015; 12: 3801-3808.
10. Wang X, Wang W, Liu C, Wu XJ. Involvement of TRPC1 and cyclin D1 in human pulmonary artery smooth muscle cells proliferation induced by cigarette smoke extract. Curr Med Sci 2020; 40: 1085-1091.
11. Groth A, Vrugt B, Brock M, Speich R, Ulrich S, Huber LC. Inflammatory cytokines in pulmonary hypertension. Respir Res 2014; 15: 47-55.
12. Shao J, Wang P, Liu A, Du X, Bai J, Chen M. Punicalagin prevents hypoxic pulmonary hypertension via anti-oxidant effects in rats. Am J Chin Med 2016; 44: 785-801.
13. Oltmanns U, Chung KF, Walters M, John M, Mitchell JA. Cigarette smoke induces IL-8, but inhibits eotaxin and RANTES release from airway smooth muscle. Respir Res 2005; 6: 74.
14. Collaborators GT. Smoking prevalence and attributable disease burden in 195 countries and territories, 1990-2015: a systematic analysis from the Global Burden of Disease Study 2015. Lancet 2017; 389: 1885-1906.
15. Wang W, Zhao T, Geng K, Yuan G, Chen Y, Xu Y. Smoking and the pathophysiology of peripheral artery disease. Front Cardiovasc Med 2021; 8: 704106.
16. Fowles J, Dybing E. Application of toxicological risk assessment principles to the chemical constituents of cigarette smoke. Tob Control 2003; 12: 424-430.
17. Blanco I, Piccari L, Barberà JA. Pulmonary vasculature in COPD: The silent component. Respirology 2016; 21: 984-994.
18. Hadzic S, Wu CY, Gredic M, Kojonazarov B, Pak O, Kraut S, et al. The effect of long-term doxycycline treatment in a mouse model of cigarette smoke-induced emphysema and pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 2021; 320: L903-l915.
19. Pietra GG, Capron F, Stewart S, Leone O, Humbert M, Robbins IM, et al. Pathologic assessment of vasculopathies in pulmonary hypertension. J Am Coll Cardiol 2004; 43: 25s-32s.
20. Rabinovitch M. The mouse through the looking glass: A new door into the pathophysiology of pulmonary hypertension. Circ Res 2004; 94: 1001-1004.
21. Hu J, Xu YJ, Zhang ZX, Tian F. Effect of cigarette smoke extract on proliferation of rat pulmonary artery smooth muscle cells and the relevant roles of protein kinase C. Chin Med J (Engl) 2007; 120: 1523-1528.
22. Xing AP, Du YC, Hu XY, Xu JY, Zhang HP, Li Y, et al. Cigarette smoke extract stimulates rat pulmonary artery smooth muscle cell proliferation via PKC-PDGFB signaling. J Biomed Biotechnol 2012; 2012: 534384.
23. Dolenc J, Šebeštjen M, Vrtovec B, Koželj M, Haddad F. Pulmonary hypertension in patients with advanced heart failure is associated with increased levels of interleukin-6. Biomarkers 2014; 19: 385-390.
24. Hashimoto-Kataoka T, Hosen N, Sonobe T, Arita Y, Yasui T, Masaki T, et al. Interleukin-6/interleukin-21 signaling axis is critical in the pathogenesis of pulmonary arterial hypertension. Proc Natl Acad Sci U S A 2015; 112: E2677-2686.
25. Matura LA, Ventetuolo CE, Palevsky HI, Lederer DJ, Horn EM, Mathai SC, et al. Interleukin-6 and tumor necrosis factor-α are associated with quality of life-related symptoms in pulmonary arterial hypertension. Ann Am Thorac Soc 2015; 12: 370-375.
26. Pugliese SC, Poth JM, Fini MA, Olschewski A, El Kasmi KC, Stenmark KR. The role of inflammation in hypoxic pulmonary hypertension: from cellular mechanisms to clinical phenotypes. Am J Physiol Lung Cell Mol Physiol 2015; 308: L229-252.
27. Soon E, Holmes AM, Treacy CM, Doughty NJ, Southgate L, Machado RD, et al. Elevated levels of inflammatory cytokines predict survival in idiopathic and familial pulmonary arterial hypertension. Circulation 2010; 122: 920-927.
28. Humbert M, Monti G, Brenot F, Sitbon O, Portier A, Grangeot-Keros L, et al. Increased interleukin-1 and interleukin-6 serum concentrations in severe primary pulmonary hypertension. Am J Respir Crit Care Med 1995; 151: 1628-1631.
29. Selimovic N, Bergh CH, Andersson B, Sakiniene E, Carlsten H, Rundqvist B. Growth factors and interleukin-6 across the lung circulation in pulmonary hypertension. Eur Respir J 2009; 34: 662-668.
30. Chaouat A, Savale L, Chouaid C, Tu L, Sztrymf B, Canuet M, et al. Role for interleukin-6 in COPD-related pulmonary hypertension. Chest 2009; 136: 678-687.
31. Bhargava A, Kumar A, Yuan N, Gewitz MH, Mathew R. Monocrotaline induces interleukin-6 mRNA expression in rat lungs. Heart Dis 1999; 1: 126-132.
32. Steiner MK, Syrkina OL, Kolliputi N, Mark EJ, Hales CA, Waxman AB. Interleukin-6 overexpression induces pulmonary hypertension. Circ Res 2009; 104: 236-244, 228p following 244.
33. Joppa P, Petrasova D, Stancak B, Tkacova R. Systemic inflammation in patients with COPD and pulmonary hypertension. Chest 2006; 130: 326-333.
34. Stevens T, Janssen PL, Tucker A. Acute and long-term TNF-alpha administration increases pulmonary vascular reactivity in isolated rat lungs. J Appl Physiol 1992; 73: 708-712.
35. Fujita M, Shannon JM, Irvin CG, Fagan KA, Cool C, Augustin A, et al. Overexpression of tumor necrosis factor-alpha produces an increase in lung volumes and pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 2001; 280: L39-49.
36. Sutendra G, Dromparis P, Bonnet S, Haromy A, McMurtry MS, Bleackley RC, et al. Pyruvate dehydrogenase inhibition by the inflammatory cytokine TNFα contributes to the pathogenesis of pulmonary arterial hypertension. J Mol Med (Berl) 2011; 89: 771-783.
37. Wang Q, Zuo XR, Wang YY, Xie WP, Wang H, Zhang M. Monocrotaline-induced pulmonary arterial hypertension is attenuated by TNF-α antagonists via the suppression of TNF-α expression and NF-κB pathway in rats. Vascul Pharmacol 2013; 58: 71-77.
38. Mutschler D, Wikström G, Lind L, Larsson A, Lagrange A, Eriksson M. Etanercept reduces late endotoxin-induced pulmonary hypertension in the pig. J Interferon Cytokine Res 2006; 26: 661-667.
39. Arnson Y, Shoenfeld Y, Amital H. Effects of tobacco smoke on immunity, inflammation and autoimmunity. J Autoimmun 2010; 34:J258-265.
40. Oeckinghaus A, Hayden MS, Ghosh S. Crosstalk in NF-κB signaling pathways. Nat Immunol 2011; 12: 695-708.
41. Li L, Wei C, Kim IK, Janssen-Heininger Y, Gupta S. Inhibition of nuclear factor-κB in the lungs prevents monocrotaline-induced pulmonary hypertension in mice. Hypertension 2014; 63: 1260-1269.
42. Ogawa A, Firth AL, Yao W, Rubin LJ, Yuan JX. Prednisolone inhibits PDGF-induced nuclear translocation of NF-kappaB in human pulmonary artery smooth muscle cells. Am J Physiol Lung Cell Mol Physiol 2008; 295: L648-657.
43. Fan J, Fan X, Li Y, Ding L, Zheng Q, Guo J, et al. Chronic Normobaric Hypoxia Induces Pulmonary Hypertension in Rats: Role of NF-κB. High Alt Med Biol 2016; 17: 43-49.
44. Obata H, Biro S, Arima N, Kaieda H, Kihara T, Eto H, et al. NF-kappa B is induced in the nuclei of cultured rat aortic smooth muscle cells by stimulation of various growth factors. Biochem Biophys Res Commun 1996; 224: 27-32.
45. Makarenko VV, Usatyuk PV, Yuan G, Lee MM, Nanduri J, Natarajan V, et al. Intermittent hypoxia-induced endothelial barrier dysfunction requires ROS-dependent MAP kinase activation. Am J Physiol Cell Physiol 2014; 306: C745-752.
46. Tuleta I, França CN, Wenzel D, Fleischmann B, Nickenig G, Werner N, et al. Hypoxia-induced endothelial dysfunction in apolipoprotein E-deficient mice; effects of infliximab and L-glutathione. Atherosclerosis 2014; 236: 400-410.
47. Aggarwal S, Gross CM, Sharma S, Fineman JR, Black SM. Reactive oxygen species in pulmonary vascular remodeling. Compr Physiol 2013; 3: 1011-1034.
48. Fulton DJR, Li X, Bordan Z, Haigh S, Bentley A, Chen F, et al. Reactive oxygen and nitrogen species in the development of pulmonary hypertension. Anti-oxidants (Basel) 2017; 6: 1-22.
49. Zuo L, Rose BA, Roberts WJ, He F, Banes-Berceli AK. Molecular characterization of reactive oxygen species in systemic and pulmonary hypertension. Am J Hypertens 2014; 27: 643-650.
50. Zhu J, Kovacs L, Han W, Liu G, Huo Y, Lucas R, et al. Reactive oxygen species-dependent calpain activation contributes to airway and pulmonary vascular remodeling in chronic obstructive pulmonary disease. Anti-oxid Redox Signal 2019; 31: 804-818.
51. Liu JQ, Zelko IN, Erbynn EM, Sham JS, Folz RJ. Hypoxic pulmonary hypertension: role of superoxide and NADPH oxidase (gp91phox). Am J Physiol Lung Cell Mol Physiol 2006; 290: L2-10.
52. Jin H, Liu M, Zhang X, Pan J, Han J, Wang Y, et al. Grape seed procyanidin extract attenuates hypoxic pulmonary hypertension by inhibiting oxidative stress and pulmonary arterial smooth muscle cells proliferation. J Nutr Biochem 2016; 36: 81-88.
53. Joppa P, Petrásová D, Stancák B, Dorková Z, Tkácová R. Oxidative stress in patients with COPD and pulmonary hypertension. Wien Klin Wochenschr 2007; 119: 428-434.
54. Li C, Yan L, Xu J. Correlations between lipid ratio/oxidative stress status in COPD patients and pulmonary hypertension as well as prognosis. Zhong Nan Da Xue Xue Bao Yi Xue Ban 2016; 41: 1168-1174.
55. Kluchová Z, Tkáčová R. The role of oxidative stress in lung injury induced by cigarette smoke. Biologia 2006; 61: 643-650.
56. Yamaguchi Y, Nasu F, Harada A, Kunitomo M. Oxidants in the gas phase of cigarette smoke pass through the lung alveolar wall and raise systemic oxidative stress. J Pharmacol Sci 2007; 103: 275-282.
57. Xiao F, Li X, Wang J, Cao J. Mechanisms of vascular endothelial cell injury in response to intermittent and/or continuous hypoxia exposure and protective effects of anti-inflammatory and anti-oxidant agents. Sleep Breath 2019; 23: 515-522.
58. Simonsen U, Christensen FH, Buus NH. The effect of tempol on endothelium-dependent vasodilatation and blood pressure. Pharmacol Ther 2009; 122: 109-124.
59. Bourgoin F, Bachelard H, Badeau M, Larivière R, Nadeau A, Pitre M. Effects of tempol on endothelial and vascular dysfunctions and insulin resistance induced by a high-fat high-sucrose diet in the rat. Can J Physiol Pharmacol 2013; 91: 547-561.
60. Hayakawa M, Miyashita H, Sakamoto I, Kitagawa M, Tanaka H, Yasuda H, et al. Evidence that reactive oxygen species do not mediate NF-kappaB activation. Embo j 2003; 22: 3356-3366.
Iranian Journal of Basic Medical Sciences
Volume 25, Issue 6
June 2022
Pages 755-761

Files

Share

How to cite

Statistics