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Article

Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2

Retraction in: /10.3892/or.2019.7087
  • Authors:
    • Jian Wu
    • An‑Quan Shang
    • Chu Chen
    • Wei‑Wei Wang
    • Cun‑Quan Xiong
    • Nai‑Zhou Guo
  • View Affiliations / Copyright

    Affiliations: Laboratory of Medicine, The First People's Hospital of Yancheng City, Yancheng, Jiangsu 224005, P.R. China, Department of Laboratory Medicine, Tongji Hospital of Tongji University, Shanghai 200092, P.R. China, Kangda College of Nanjing Medical University, Lianyungang, Jiangsu 222000, P.R. China, Department of Laboratory Medicine, The Sixth People's Hospital of Yancheng City, Yancheng, Jiangsu 224005, P.R. China, College of Pharmacy, Jiangsu Vocational College Medicine, Yancheng, Jiangsu 224002, P.R. China
  • Pages: 3033-3039
    |
    Published online on: August 17, 2018
       https://doi.org/10.3892/or.2018.6656
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Abstract

The aim of the present study was to investigate the mechanisms and protective effect of serum‑free‑medium‑type fetal placental mesenchymal stem cell (fPMSC) culture supernatant on A549 lung epithelial cells following treatment with hydrogen peroxide (H2O2). A549 lung epithelial cells were stimulated with different concentrations of H2O2, and the survival rate of the cells was examined by Cell Counting Kit‑8 (CCK‑8) assay. It was concluded that the H2O2 concentration when the cell survival rate was at 50% was the optimum condition to create an oxidative damage model. Hoechst 33258 staining and western blot analysis was used to validate the A549 lung epithelial cell model. Serum‑free medium was used to culture fPMSCs, and A549 lung epithelial cells treated with H2O2 were cultured with passage 3 MSC supernatant for 24 h. This was termed the supernatant group. Simultaneously, a damage group that was stimulated with H2O2 only, and a vitamin C (VC) group that was treated with H2O2 followed by 100 µmol/l VC in culture medium was also established. The apoptosis of the three groups was detected by flow cytometry, and western blotting was used to detect apoptosis‑associated and nuclear factor erythroid 2‑like 2 (Nrf2)‑kelch‑like ECH‑associated protein 1 (Keap1)‑antioxidant response element/oxidative stress‑associated protein expression. Following the CCK‑8 test, 600 µmol/l H2O2 was selected to stimulate the A549 lung epithelial cells for 24 h, which resulted in a A549 cell survival rate of 56.41±3.31%. Hoechst 33258 staining and western blotting also confirmed the reliability of the model. Flow cytometry demonstrated that the apoptotic rate of the cells in the VC and supernatant groups was reduced compared with that in the injury group. The difference between the supernatant group and the injury group was statistically significant. The detection of apoptosis‑associated proteins by western blotting revealed that the expression of apoptosis regulator BAX and Caspase‑3 in the VC and supernatant groups was decreased. Furthermore, the expression of B‑cell lymphoma‑2 was increased compared with that in the injury group, and the difference was statistically significant (P<0.05). Compared with that in the injury group, the expression of Nrf2 increased in the VC and supernatant groups, whereas the expression of Keap1 was decreased, and the difference was statistically significant (P<0.05). In conclusion, fPMSC supernatant exhibited an antioxidant capacity in A549 lung epithelial cells treated with H2O2 as a model of acute lung injury. The supernatant was found to reduce oxidative damage and inhibit apoptosis.
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View References

1 

Jin Z, Suen Chun K and Ma D: Perioperative ‘remote’ acute lung injury: Recent update. J Biomed Res. 31:197–212. 2017.PubMed/NCBI

2 

Hayes M, Curley G, Ansari B and Laffey JG: Clinical review: Stem cell therapies for acute lung injury/acute respiratory distress syndrome - hope or hype? Crit Care. 16:2052012. View Article : Google Scholar : PubMed/NCBI

3 

Han F, Luo Y, Li Y, Liu Z, Xu D, Jin F and Li Z: Seawater induces apoptosis in alveolar epithelial cells via the Fas/FasL-mediated pathway. Respir Physiol Neurobiol. 182:71–80. 2012. View Article : Google Scholar : PubMed/NCBI

4 

Al-Biltagi MA, Abo-Elezz AA, Elshafiey RM, Suliman GA, Mabrouk MM and Mourad HA: The predictive value of soluble endothelial selectin plasma levels in children with acute lung injury. J Crit Care. 32:31–35. 2016. View Article : Google Scholar : PubMed/NCBI

5 

Trajano Lima ET, Sternberg C, Caetano M, Silva Santos MA, Porto LC, Santos JC, Ribeiro ML, Magalhães CB, Zin WA, Benjamim CF, et al: Endotoxin-induced acute lung injury is dependent upon oxidative response. Inhal Toxicol. 23:918–926. 2011. View Article : Google Scholar : PubMed/NCBI

6 

Poljsak B, Suput D and Milisav I: Achieving the balance between ROS and antioxidants: when to use the synthetic antioxidants. Oxid Med Cell Longev. 2013:9567922013. View Article : Google Scholar : PubMed/NCBI

7 

Yang C, Jiang J, Yang X, Wang H and Du J: Stem/progenitor cells in endogenous repairing responses: New toolbox for the treatment of acute lung injury. J Transl Med. 14:472016. View Article : Google Scholar : PubMed/NCBI

8 

Miller DM, Singh IN, Wang JA and Hall ED: Administration of the Nrf2-ARE activators sulforaphane and carnosic acid attenuates 4-hydroxy-2-nonenal-induced mitochondrial dysfunction ex vivo. Free Radic Biol Med. 57:1–9. 2013. View Article : Google Scholar : PubMed/NCBI

9 

Suniara RK, Jenkinson EJ and Owen JJ: An essential role for thymic mesenchyme in early T cell development. J Exp Med. 191:1051–1056. 2000. View Article : Google Scholar : PubMed/NCBI

10 

Shalaby SM, El-Shal AS, Abd-Allah SH, Selim AO, Selim SA, Gouda ZA, El Motteleb Abd DM, Zanfaly HE, El-Assar HM and Abdelazim S: Mesenchymal stromal cell injection protects against oxidative stress in Escherichia coli-induced acute lung injury in mice. Cytotherapy. 16:764–775. 2014. View Article : Google Scholar : PubMed/NCBI

11 

Wilson JG, Liu KD, Zhuo H, Caballero L, McMillan M, Fang X, Cosgrove K, Vojnik R, Calfee CS, Lee JW, et al: Mesenchymal stem (stromal) cells for treatment of ARDS: A phase 1 clinical trial. Lancet Respir Med. 3:24–32. 2015. View Article : Google Scholar : PubMed/NCBI

12 

Castro-Manrreza ME and Montesinos JJ: Immunoregulation by mesenchymal stem cells: Biological aspects and clinical applications. J Immunol Res. 2015:3949172015. View Article : Google Scholar : PubMed/NCBI

13 

Zhang G, Zou X, Huang Y, Wang F, Miao S, Liu G, Chen M and Zhu Y: Mesenchymal stromal cell-derived extracellular vesicles protect against acute kidney injury through anti-oxidation by enhancing Nrf2/ARE activation in Rats. Kidney Blood Press Res. 41:119–128. 2016. View Article : Google Scholar : PubMed/NCBI

14 

Pleyer L, Valent P and Greil R: Mesenchymal stem and progenitor cells in normal and dysplastic hematopoiesis-masters of survival and clonality? Int J Mol Sci. 17:10092016. View Article : Google Scholar :

15 

Lv H, Liu Q, Wen Z, Feng H, Deng X and Ci X: Xanthohumol ameliorates lipopolysaccharide (LPS)-induced acute lung injury via induction of AMPK/GSK3β-Nrf2 signal axis. Redox Biol. 12:311–324. 2017. View Article : Google Scholar : PubMed/NCBI

16 

Ware LB and Matthay MA: The acute respiratory distress syndrome. N Engl J Med. 141:460–470. 2000.

17 

Keum YS and Choi BY: Molecular and chemical regulation of the Keap1-Nrf2 signaling pathway. Molecules. 19:10074–10089. 2014. View Article : Google Scholar : PubMed/NCBI

18 

Wang Z, Ji C, Wu L, Qiu J, Li Q, Shao Z and Chen G: Tert-butylhydroquinone alleviates early brain injury and cognitive dysfunction after experimental subarachnoid hemorrhage: Role of Keap1/Nrf2/ARE pathway. PLoS One. 9:e976852014. View Article : Google Scholar : PubMed/NCBI

19 

Liu X, Zheng P, Xiaodong G, Guanghui D, Hongbin CH, Zan ZH, Rongrong H, Xinxin N, Jing SH and Yihua A: A preliminary evaluation of efficacy and safety of Wharton's jelly mesenchymal stem cell transplantation in patients with type 2 diabetes mellitus. Stem Cell Research & Therapy. 5:1–9. 2014. View Article : Google Scholar

20 

Jang YO, Kim YJ, Baik SK, Kim MY, Eom YW, Cho MY, Park HJ, Park SY, Kim BR, Kim JW, et al: Histological improvement following administration of autologous bone marrow-derived mesenchymal stem cells for alcoholic cirrhosis: A pilot study. Liver Int. 34:33–41. 2014. View Article : Google Scholar : PubMed/NCBI

21 

Hare JM, Fishman JE, Gerstenblith G, DiFede Velazquez DL, Zambrano JP, Suncion VY, Tracy M, Ghersin E, Johnston PV, Brinker JA, Breton E, et al: Comparison of allogeneic vs autologous bone marrow-derived mesenchymal stem cells delivered by transendocardial injection in patients with ischemic cardiomyopathy. JAMA. 308:2369–2379. 2012. View Article : Google Scholar : PubMed/NCBI

22 

Ni S, Wang D, Qiu X, Pang L, Song Z and Guo K: Bone marrow mesenchymal stem cells protect against bleomycin-induced pulmonary fibrosis in rat by activating Nrf2 signaling. Int J Clin Exp Pathol. 8:7752–7761. 2015.PubMed/NCBI

23 

Chang YS, Ahn SY, Yoo HS, Sung SI, Choi SJ, Oh WI and Park WS: Mesenchymal stem cells for bronchopulmonary dysplasia: phase 1 dose-escalation clinical trial. J Pediatr. 164:966–972. 2014. View Article : Google Scholar : PubMed/NCBI

24 

Haddad R and Saldanha-Araujo F: Mechanisms of T-cell immunosuppression by mesenchymal stromal cells: What do we know so far? BioMed Res Int. 2014:2168062014. View Article : Google Scholar : PubMed/NCBI

25 

Martinet L, Fleury-Cappellesso S, Gadelorge M, Dietrich G, Bourin P, Fournié JJ and Poupot R: A regulatory cross-talk between Vγ9Vδ2 T lymphocytes and mesenchymal stem cells. Eur J Immunol. 39:752–762. 2009. View Article : Google Scholar : PubMed/NCBI

26 

Tasso R, Augello A, Carida' M, Postiglione F, Tibiletti MG, Bernasconi B, Astigiano S, Fais F, Truini M, Cancedda R, et al: Development of sarcomas in mice implanted with mesenchymal stem cells seeded onto bioscaffolds. Carcinogenesis. 30:150–157. 2009. View Article : Google Scholar : PubMed/NCBI

27 

Chen W, Huang Y, Han J, Yu L, Li Y, Lu Z, Li H, Liu Z, Shi C, Duan F, et al: Immunomodulatory effects of mesenchymal stromal cells-derived exosome. Immunol Res. 64:831–840. 2016. View Article : Google Scholar : PubMed/NCBI

28 

Sabin K and Kikyo N: Microvesicles as mediators of tissue regeneration. Transl Res. 163:286–295. 2014. View Article : Google Scholar : PubMed/NCBI

29 

Sdrimas K and Kourembanas S: MSC microvesicles for the treatment of lung disease: a new paradigm for cell-free therapy. Antioxid Redox Signal. 21:1905–1915. 2014. View Article : Google Scholar : PubMed/NCBI

30 

Hashemian SJ, Kouhnavard M and Nasli-Esfahani E: Mesenchymal stem cells: Rising concerns over their application in treatment of type one diabetes mellitus. J Diabetes Res. 2015:6751032015. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Wu J, Shang AQ, Chen C, Wang WW, Xiong CQ and Guo NZ: Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087. Oncol Rep 40: 3033-3039, 2018.
APA
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., & Guo, N. (2018). Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087. Oncology Reports, 40, 3033-3039. https://doi.org/10.3892/or.2018.6656
MLA
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., Guo, N."Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087". Oncology Reports 40.5 (2018): 3033-3039.
Chicago
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., Guo, N."Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087". Oncology Reports 40, no. 5 (2018): 3033-3039. https://doi.org/10.3892/or.2018.6656
Copy and paste a formatted citation
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Spandidos Publications style
Wu J, Shang AQ, Chen C, Wang WW, Xiong CQ and Guo NZ: Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087. Oncol Rep 40: 3033-3039, 2018.
APA
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., & Guo, N. (2018). Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087. Oncology Reports, 40, 3033-3039. https://doi.org/10.3892/or.2018.6656
MLA
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., Guo, N."Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087". Oncology Reports 40.5 (2018): 3033-3039.
Chicago
Wu, J., Shang, A., Chen, C., Wang, W., Xiong, C., Guo, N."Serum‑free‑medium‑type mesenchymal stem cell culture supernatant exerts a protective effect on A549 lung epithelial cells in acute lung injury induced by H2O2 Retraction in /10.3892/or.2019.7087". Oncology Reports 40, no. 5 (2018): 3033-3039. https://doi.org/10.3892/or.2018.6656
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