https://doi.org/10.4081/ejh.2023.3535
LncRNA gadd7 promotes mitochondrial membrane potential decrease and apoptosis of alveolar type II epithelial cells by positively regulating MFN1 in an in vitro model of hyperoxia-induced acute lung injury
Submitted: 16 August 2022
Accepted: 17 April 2023
Published: 31 May 2023
Accepted: 17 April 2023
Abstract Views: 502
PDF: 406
HTML: 15
HTML: 15
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Authors
Hyperoxia-induced acute lung injury (HALI) is a complication of oxygen therapy. LncRNA gadd7 shows associations with HALI. We explored the effects of gadd7 on mitochondrial membrane potential (MMP) and apoptosis of alveolar type II epithelial cells (AEC II) in HALI. RLE-6TN cells were transfected with 50 nM silenced and overexpressed gadd7 and MFN1 plasmids (sh-gadd7, oe-gadd7 and/or oe-MFN1), respectively for 48 h, followed by hyperoxic culture for 24 h. gadd7 and MFN1 levels were assessed by RT-qPCR and Western blot. RLE-6TN cells were stained by JC-1 and MMP changes were observed using a fluorescence microscope. The average fluorescence intensity of red and green fluorescence was evaluated with the help of Image J software, and the ratio of red/green fluorescence intensity represented MMP. Cell apoptosis was assessed by flow cytometry using Annexin V-FITC/PI double staining method. The levels of apoptosis proteins [Bax, Bcl-2, Cleaved caspase-3, Cleaved poly ADP-ribose polymerase (PARP)] were measured by Western blot. LncRNA gadd7 and MFN1 were highly expressed in hyperoxia-induced AEC II. Hyperoxia induced MMP decrease and apoptosis promotion in RLE-6TN cells. LncRNA gadd7 positively regulated MFN1 expression. Knockdown of gadd7 inhibited MMP decrease and apoptosis of hyperoxia-induced RLE-6TN cells. MFN1 overexpression annulled the inhibitory effects of gadd7 silencing on MMP decrease and apoptosis in RLE-6TN cells. Briefly, lncRNA gadd7 promoted MMP decrease and apoptosis of AEC II by positively regulating the expression of MFN1 in the HALI model in vitro.
Jiang J, Wang J, Li C, Mo L, Huang D. Hyperoxia induces alveolar epithelial cell apoptosis by regulating mitochondrial function through small mothers against decapentaplegic 3 (SMAD3) and extracellular signal-regulated kinase 1/2 (ERK1/2). Bioengineered 2022;13:242-52.
DOI: https://doi.org/10.1080/21655979.2021.2012953
Wu D, Liang M, Dang H, Fang F, Xu F, Liu C. Hydrogen protects against hyperoxia-induced apoptosis in type II alveolar epithelial cells via activation of PI3K/Akt/Foxo3a signaling pathway. Biochem Biophys Res Commun 2018;495:1620-7.
DOI: https://doi.org/10.1016/j.bbrc.2017.11.193
Jacobs SS, Krishnan JA, Lederer DJ, Ghazipura M, Hossain T, Tan AM, et al. Home oxygen therapy for adults with chronic lung disease. An Official American Thoracic Society Clinical Practice Guideline. Am J Respir Crit Care Med 2020;202:e121-41.
DOI: https://doi.org/10.1164/rccm.202009-3608ST
Wenger HC, Cifu AS, Lee CT. Home oxygen therapy for adults with chronic obstructive pulmonary disease or interstitial lung disease. JAMA 2021;326:1738-9.
DOI: https://doi.org/10.1001/jama.2021.12073
Zhu X, Lei X, Wang J, Dong W. Protective effects of resveratrol on hyperoxia-induced lung injury in neonatal rats by alleviating apoptosis and ROS production. J Matern Fetal Neonatal Med 2020;33:4150-8.
DOI: https://doi.org/10.1080/14767058.2019.1597846
Qin S, Wang H, Liu G, Mei H, Chen M. miR215p ameliorates hyperoxic acute lung injury and decreases apoptosis of AEC II cells via PTEN/AKT signaling in rats. Mol Med Rep 2019;20:4953-62.
DOI: https://doi.org/10.3892/mmr.2019.10779
Ao X, Fang F, Xu F. Vasoactive intestinal peptide protects alveolar epithelial cells against hyperoxia via promoting the activation of STAT3. Regul Pept 2011;168:1-4.
DOI: https://doi.org/10.1016/j.regpep.2011.02.006
Lee HS, Kim C K. Effect of recombinant IL-10 on cultured fetal rat alveolar type II cells exposed to 65%-hyperoxia. Respir Res 2011;12:68.
DOI: https://doi.org/10.1186/1465-9921-12-68
Li XG, Song X, Wang JY, Sun CH, Li ZQ, Meng LL, et al. Fibroblast growth factor 18 alleviates hyperoxia-induced lung injury in mice by adjusting oxidative stress and inflammation. Eur Rev Med Pharmacol Sci 2021;25:1485-94.
Lu X, Wang C, Wu D, Zhang C, Xiao C, Xu F. Quantitative proteomics reveals the mechanisms of hydrogen-conferred protection against hyperoxia-induced injury in type II alveolar epithelial cells. Exp Lung Res 2018;44:464-75.
DOI: https://doi.org/10.1080/01902148.2019.1601296
Ratner V, Starkov A, Matsiukevich D, Polin RA, Ten V S. Mitochondrial dysfunction contributes to alveolar developmental arrest in hyperoxia-exposed mice. Am J Respir Cell Mol Biol 2009;40:511-8.
DOI: https://doi.org/10.1165/rcmb.2008-0341RC
Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC. Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J Cell Biol 2003;160:189-200.
DOI: https://doi.org/10.1083/jcb.200211046
Sidarala V, Zhu J, Levi-D'Ancona E, Pearson GL, Reck EC, Walker EM, et al. Mitofusin 1 and 2 regulation of mitochondrial DNA content is a critical determinant of glucose homeostasis. Nat Commun 2022;13:2340.
DOI: https://doi.org/10.1038/s41467-022-29945-7
Rovira-Llopis S, Banuls C, Diaz-Morales N, Hernandez-Mijares A, Rocha M, Victor VM. Mitochondrial dynamics in type 2 diabetes: Pathophysiological implications. Redox Biol 2017;11:637-45.
DOI: https://doi.org/10.1016/j.redox.2017.01.013
Chung KP, Hsu CL, Fan LC, Huang Z, Bhatia D, Chen YJ, et al. Mitofusins regulate lipid metabolism to mediate the development of lung fibrosis. Nat Commun 2019;10:3390.
DOI: https://doi.org/10.1038/s41467-019-11327-1
Liu G, Mei H, Chen M, Qin S, Li K, Zhang W, et al. Protective effect of agmatine against hyperoxia-induced acute lung injury via regulating lncRNA gadd7. Biochem Biophys Res Commun 2019;516:68-74.
DOI: https://doi.org/10.1016/j.bbrc.2019.04.164
Mu G, Deng Y, Lu Z, Li X, Chen Y. miR-20b suppresses mitochondrial dysfunction-mediated apoptosis to alleviate hyperoxia-induced acute lung injury by directly targeting MFN1 and MFN2. Acta Bioch Bioph Sin 2021;53:220-8.
DOI: https://doi.org/10.1093/abbs/gmaa161
Zhang L, Wang P, Shen Y, Huang T, Hu X, Yu W. Mechanism of lncRNA H19 in regulating pulmonary injury in hyperoxia-induced bronchopulmonary dysplasia newborn mice. Am J Perinatol 2022;39:1089-96.
DOI: https://doi.org/10.1055/s-0040-1721498
Mei M, Nie J, Sun H, Wang H, Rong L. LncRNA-NEF regulated the hyperoxia-induced injury of lung epithelial cells by FOXA2. Am J Transl Res 2020;12:5563-74.
Ji L, Liu Z, Dong C, Wu D, Yang S, Wu L. LncRNA CASC2 targets CAV1 by competitively binding with microRNA-194-5p to inhibit neonatal lung injury. Exp Mol Pathol 2021;118:104575.
DOI: https://doi.org/10.1016/j.yexmp.2020.104575
Liu X, Li D, Zhang W, Guo M, Zhan Q. Long non-coding RNA gadd7 interacts with TDP-43 and regulates Cdk6 mRNA decay. EMBO J 2012;31:4415-27.
DOI: https://doi.org/10.1038/emboj.2012.292
Zhao J, Ma W, Zhong Y, Deng H, Zhou B, Wu Y, et al. Transcriptional inhibition of lncRNA gadd7 by CRISPR/dCas9-KRAB protects spermatocyte viability. Front Mol Biosci 2021;8:652392.
DOI: https://doi.org/10.3389/fmolb.2021.652392
Zhao J, Li H, Deng H, Zhu L, Zhou B, Yang M, et al. LncRNA gadd7, increased in varicocele patients, suppresses cell proliferation and promotes cell apoptosis. Oncotarget 2018;9:5105-10.
DOI: https://doi.org/10.18632/oncotarget.23696
Luo Y, Hao T, Zhang J, Zhang M, Sun P, Wu L. MicroRNA-592 suppresses the malignant phenotypes of thyroid cancer by regulating lncRNA NEAT1 and downregulating NOVA1. Int J Mol Med 2019;44:1172-82.
DOI: https://doi.org/10.3892/ijmm.2019.4278
Zaib S, Hayyat A, Ali N, Gul A, Naveed M, Khan I. Role of mitochondrial membrane potential and lactate dehydrogenase A in apoptosis. Anticancer Agents Med Chem 2022;22:2048-62.
DOI: https://doi.org/10.2174/1871520621666211126090906
Sidramagowda Patil S, Hernandez-Cuervo H, Fukumoto J, Krishnamurthy S, Lin M, Alleyn M, et al. Alda-1 attenuates hyperoxia-induced acute lung injury in mice. Front Pharmacol 2020;11:597942.
DOI: https://doi.org/10.3389/fphar.2020.597942
Wu Y, Zhang Z, Li J, Zhong H, Yuan R, Deng Z, et al. Mechanism of adipose-derived mesenchymal stem cell-derived extracellular vesicles carrying miR-21-5p in hyperoxia-induced lung injury. Stem Cell Rev Rep 2022;18:1007-24.
DOI: https://doi.org/10.1007/s12015-021-10311-x
Zou DM, Zhou SM, Li LH, Zhou JL, Tang ZM, Wang SH. Knockdown of long noncoding RNAs of maternally expressed 3 alleviates hyperoxia-induced lung injury via inhibiting thioredoxin-interacting protein-mediated pyroptosis by binding to miR-18a. Am J Pathol 2020;190:994-1005.
DOI: https://doi.org/10.1016/j.ajpath.2019.12.013
Zhu X, Wang F, Lei X, Dong W. Resveratrol alleviates alveolar epithelial cell injury induced by hyperoxia by reducing apoptosis and mitochondrial dysfunction. Exp Biol Med (Maywood) 2021;246:596-606.
DOI: https://doi.org/10.1177/1535370220975106
Zhu Y, Ju H, Lu H, Tang W, Lu J, Wang Q. The function role of ubiquitin proteasome pathway in the ER stress-induced AECII apoptosis during hyperoxia exposure. BMC Pulm Med 2021;21:379.
DOI: https://doi.org/10.1186/s12890-021-01751-9
Lu HY, Zhang J, Wang QX, Tang W, Zhang LJ. Activation of the endoplasmic reticulum stress pathway involving CHOP in the lungs of rats with hyperoxia induced bronchopulmonary dysplasia. Mol Med Rep 2015;12:4494-500.
DOI: https://doi.org/10.3892/mmr.2015.3979
Oanh NTK, Park YY, Cho H. Mitochondria elongation is mediated through SIRT1-mediated MFN1 stabilization. Cell Signal 2017;38:67-75.
DOI: https://doi.org/10.1016/j.cellsig.2017.06.019
Park YY, Nguyen OT, Kang H, Cho H. MARCH5-mediated quality control on acetylated Mfn1 facilitates mitochondrial homeostasis and cell survival. Cell Death Dis 2014;5:e1172.
DOI: https://doi.org/10.1038/cddis.2014.142
Ethics Approval
The experiments were authorized by the Academic Ethics Committee of The Second Affiliated Hospital of Zunyi Medical UniversitySupporting Agencies
National Natural Science Foundation of China , Science and Technology Project of Guizhou ProvinceHow to Cite
Liu, G., Yin, C., Qian, M., Xiao, X., Wu, H., & Fu, F. (2023). LncRNA gadd7 promotes mitochondrial membrane potential decrease and apoptosis of alveolar type II epithelial cells by positively regulating MFN1 in an <em>in vitro</em> model of hyperoxia-induced acute lung injury. European Journal of Histochemistry, 67(2). https://doi.org/10.4081/ejh.2023.3535
Copyright (c) 2023 The Author(s)
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
PAGEPress has chosen to apply the Creative Commons Attribution NonCommercial 4.0 International License (CC BY-NC 4.0) to all manuscripts to be published.
