Effects of mechanical stretch on fiber proliferation in human lung epithelial cells

doi:10.12307/2022.560

Abstract

Abstract: BACKGROUND: Mechanical ventilation is currently one of important and effective treatment and support methods for severe respiratory diseases, but improper mechanical ventilation can aggravate lung injury and even form fibrosis. How to minimize the lung damage associated with the ventilator and play real protective role of the ventilator is very important. The lung is a mechanical organ. Mechanical ventilation causes the alveoli to be inflated repeatedly, which stretches the neighboring lung epithelial cells, and puts it in an abnormal damage repair. Therefore, excessive mechanical stretch may play an important role in promoting the occurrence of lung fibrosis. At present, there are few reports on the effect of mechanical stretch on the proliferation of lung epithelial cells at home and abroad, and the specific mechanism is not clear.
OBJECTIVE: To investigate the effect of different mechanical stretch strengths and stretch time on expression of transforming growth factor β1, vimentin, and type I collagen in human lung epithelial BEAS-2B cells.
METHODS: Human lung epithelial BEAS-2B cells were cultured in vitro. Cells at logarithmic growth phase were divided into control static group, 10% stretch group, and 20% stretch group. The FX-5000 system was used to stretch BEAS-2B cells at 20 times/min and sine wave for 24, 48, and 72 hours. The morphological changes in cells were observed with the inverted microscope. The expression levels of transforming growth factor β1, vimentin, and type I collagen mRNA were evaluated by RT-qPCR and immunofluorescence.
RESULTS AND CONCLUSION: (1) BEAS-2B cells displayed cobblestone morphology and linked closely in the control static group. The cell morphology changed from cobblestone shape into long spindle, and intercellular space increased obviously after 20% mechanical stretch for 72 hours. (2) The expression of mRNA and protein of transforming growth factor β1, vimentin, and type I collagen in human lung epithelial BEAS-2B cells was up-regulated with increasing stretch force in a time-dependent manner compared with the control static group (P < 0.05). Compared with the control static group, the changes in transforming growth factor β1, vimentin, and type I collagen were not significant in the 10% stretch group. (3) It is concluded that mechanical stretch increases the expression of transforming growth factor β1, vimentin, and type I collagen in human lung epithelial BEAS-2B cells. Excessive mechanical stretch plays an important role in promoting lung fiber proliferation.

Key words: mechanical stretch, lung epithelial cells, pulmonary fibrosis, transforming growth factor β1, vimentin

CLC Number:

Zhang Rong, Liang Zhenting, Yang Chun, Liu Dongdong, Xi Yin, Zhang Jie, Wang Ya, Xu Yonghao, Liu Xiaoqing, Li Yimin. Effects of mechanical stretch on fiber proliferation in human lung epithelial cells[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(24): 3821-3825.

Figures/Tables 4

References

[1] MARINI JJ, ROCCO PRM, GATTINONI L. Static and Dynamic Contributors to Ventilator-induced Lung Injury in Clinical Practice. Pressure, Energy, and Power. Am J Respir Crit Care Med. 2020;201(7):767-774.
[2] GAVER DP 3RD, NIEMAN GF, GATTO LA, et al. The POOR Get POORer: A Hypothesis for the Pathogenesis of Ventilator-induced Lung Injury. Am J Respir Crit Care Med. 2020;202(8):1081-1087.
[3] BURNHAM EL, JANSSEN WJ, RICHES DW, et al. The fibroproliferative response in acute respiratory distress syndrome: mechanisms and clinical significance. Eur Respir J. 2014;43(1):276-285.
[4] NEWTON CA, HERZOG EL. Molecular Markers and the Promise of Precision Medicine for Interstitial Lung Disease. Clin Chest Med. 2021; 42(2):357-364.
[5] JAIN M. Fibroproliferative ARDS in the Era of low-tidal-volume ventilation. Chest. 2014;146(5):1140-1142.
[6] KIM KK, SHEPPARD D, CHAPMAN HA. TGF-β1 Signaling and Tissue Fibrosis. Cold Spring Harb Perspect Biol. 2018;10(4):a022293.
[7] FROESE AR, SHIMBORI C, BELLAYE PS, et al. Stretch-induced Activation of Transforming Growth Factor-beta1 in Pulmonary Fibrosis. Am J Respir Crit Care Med. 2016;194(1):84-96.
[8] LODYGA M, HINZ B. TGF-β1 - A truly transforming growth factor in fibrosis and immunity. Semin Cell Dev Biol. 2020;101:123-139.
[9] BAI L, LI A, GONG C, et al. Protective effect of rutin against bleomycin induced lung fibrosis: Involvement of TGF-β1/α-SMA/Col I and III pathway. Biofactors. 2020;46(4):637-644.
[10] ROCCO PRM, MARINI JJ. What have we learned from animal models of ventilator-induced lung injury? Intensive Care Med. 2020;46(12):2377-2380.
[11] SLUTSKY AS. History of Mechanical Ventilation. From Vesalius to Ventilator-induced Lung Injury. Am J Respir Crit Care Med. 2015; 191(10):1106-1115.
[12] JAMAATI H, NAZARI M, DAROOEI R, et al. Role of shear stress in ventilator-induced lung injury. Lancet Respir Med. 2016;4(8):e41-e42.
[13] PAPAZIAN L, DODDOLI C, CHETAILLE B, et al. A contributive result of open-lung biopsy improves survival in acute respiratory distress syndrome patients. Crit Care Med. 2007;35(3):755-762.
[14] MARTIN C, PAPAZIAN L, PAYAN MJ, et al. Pulmonary fibrosis correlates with outcome in adult respiratory distress syndrome. A study in mechanically ventilated patients. Chest. 1995;107(1):196-200.
[15] TSCHUMPERLIN DJ, OSWARI J, MARGULIES AS. Deformation-induced injury of alveolar epithelial cells. Effect of frequency, duration, and amplitude. Am J Respir Crit Care Med. 2000;162(2 Pt 1):357-362.
[16] ZHANG R, PAN Y, FANELLI V, et al. Mechanical Stress and the Induction of Lung Fibrosis via the Midkine Signaling Pathway. Am J Respir Crit Care Med. 2015;192(3):315-323.
[17] DING Z, YUAN J, LIANG Y, et al. Ryanodine Receptor Type 2 Plays a Role in the Development of Cardiac Fibrosis under Mechanical Stretch Through TGFβ-1. Int Heart J. 2017;58(6):957-961.
[18] HAMZEH MT, SRIDHARA R, ALEXANDER LD. Cyclic stretch-induced TGF-β1 and fibronectin expression is mediated by β1-integrin through c-Src- and STAT3-dependent pathways in renal epithelial cells. Am J Physiol Renal Physiol. 2015;308(5):F425-436.
[19] CHAPMAN HA, WEI Y, MONTAS G, et al. Reversal of TGFβ1-Driven Profibrotic State in Patients with Pulmonary Fibrosis. N Engl J Med. 2020;382(11):1068-1070.
[20] ZHANG Y, DAI Y, RAMAN A, et al. Overexpression of TGF-β1 induces renal fibrosis and accelerates the decline in kidney function in polycystic kidney disease. Am J Physiol Renal Physiol. 2020;319(6):F1135-F1148.
[21] REN H, ZUO S, HOU Y, et al. Inhibition of α1-adrenoceptor reduces TGF-β1-induced epithelial-to-mesenchymal transition and attenuates UUO-induced renal fibrosis in mice. FASEB J. 2020;34(11):14892-14904.
[22] ANDUGULAPATI SB, GOURISHETTI K, TIRUNAVALLI SK, et al. Biochanin-A ameliorates pulmonary fibrosis by suppressing the TGF-β mediated EMT, myofibroblasts differentiation and collagen deposition in in vitro and in vivo systems. Phytomedicine. 2020;78:153298.
[23] GONZÁLEZ-LÓPEZ A, ALBAICETA GM. Repair after acute lung injury: molecular mechanisms and therapeutic opportunities. Crit Care. 2012; 16(2):209.
[24] GONZÁLEZ-LÓPEZ A, ASTUDILLO A, GARCÍA-PRIETO E, et al. Inflammation and matrix remodeling during repair of ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2011;301(4):L500-509.
[25] CHAUHAN C, ROTH KA. Understanding Lung Development, Injury, and Repair. Am J Pathol. 2016;186(10):2518.
[26] WALLACH-DAYAN SB, ROJAS M. Senescence, the Janus of Lung Injury and Repair. Am J Respir Cell Mol Biol. 2020;62(5):548-549.
[27] ZHAO Y, RIDGE K, ZHAO J. Acute Lung Injury, Repair, and Remodeling: Pulmonary Endothelial and Epithelial Biology. Mediators Inflamm. 2017;2017:9081521.
[28] CHANDA D, OTOUPALOVA E, SMITH SR, et al. Developmental pathways in the pathogenesis of lung fibrosis. Mol Aspects Med. 2019;65:56-69.
[29] PELOSI P, ROCCO PR. Effects of mechanical ventilation on the extracellular matrix. Intensive Care Med. 2008;34(4):631-639.
[30] BREW N, HOOPER SB, ALLISON BJ, et al. Injury and repair in the very immature lung following brief mechanical ventilation. Am J Physiol Lung Cell Mol Physiol. 2011;301(6):L917-926.
[31] BITTERMAN PB. Pathogenesis of fibrosis in acute lung injury. Am J Med. 1992;92(6A):39S-43S.
[32] STRIETER RM, MEHRAD B. New mechanisms of pulmonary fibrosis. Chest. 2009;136(5):1364-1370.
[33] WILLIS BC, DUBOIS RM, BOROK Z. Epithelial origin of myofibroblasts during fibrosis in the lung. Proc Am Thorac Soc. 2006;3(4):377-382.
[34] PARAMBIL JG, MYERS JL, AUBRY MC, et al. Causes and prognosis of diffuse alveolar damage diagnosed on surgical lung biopsy. Chest. 2007; 132(1):50-57.