[Frontiers in Bioscience, Landmark, 20, 139-156, January 1, 2015]

Isoflurane attenuates LPS-induced acute lung injury by targeting miR-155-HIF1-alpha

Rong Hu 1 , Ying Zhang 1 , Xiaohua Yang 2 , Jia Yan 1 , Yu Sun 1 , Zhifeng Chen 1 , Hong Jiang 1

1Department of Anesthesiology, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University, School of Medicine, No. 639, Zhi Zao Ju Road, Shanghai, China, 2Central Laboratory, Shanghai Chest Hospital affiliated to Shanghai Jiao Tong University

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Cell culture and treatment
    3.2. CCK8 assay
    3.3. Plasmid construction
    3.4. Luciferase reporter assays
    3.5. Transfection with HIF-1α siRNA
    3.6. Gene transfection with miR-155, miR-155 EV, miR-155 inhibitor, and miR-155 inhibitor EV
    3.7. Animals and experimental protocols
    3.8. Quantitative real-time PCR (Q RT-PCR)
    3.9. Western blot analysis
    3.10. Histopathology
    3.11. IL-1β and TNF-α level in bronchoalveolar lavage fluid
    3.12. Immunohistochemistry
    3.13. Statistical Analysis
4. Results
    4.1. Isoflurane alleviates LPS-induced cell injury
    4.2. MiR-155 and HIF-1α are involved with the isoflurane effect on LPS-induced cell injury
    4.3. MiR-155 targets the HIF-1α gene
    4.4. MiR-155 negatively regulates the expression of HIF-1α
    4.5. HO-1 is involved in the effect of isoflurane in vitro
    4.6. Postconditioning with isoflurane attenuates LPS-induced ALI
    4.7. Inverse expression of HIF1-ALPHA and miR-155 after postconditioning with isoflurane
    4.8. Overexpression of miR-155 impaired isoflurane protection against LPS-induced injury
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

Isoflurane alleviates the inflammatory response in endotoxin-induced acute lung injury (ALI). In this study, we investigated the protective mechanism of isoflurane postconditioning in lipopolysaccharide (LPS)-induced ALI. Exposure to isoflurane decreased miR-155 and upregulated HIF-1 alpha and HO-1 mRNA and protein. The effects of isoflurane on HIF-1 alpha mRNA and protein could be inhibited by overexpression of miR-155. Furthermore, mice overexpressing miR-155 had higher levels of TNF-alpha and IL-1 beta in BALF when exposed to isoflurane after LPS challenge. Conversely, downregulation of miR-155 promoted isoflurane effects on HIF-1 alpha expression. These results suggest that isoflurane posttreatment at 1 MAC for 4 hour alleviates LPS-induced ALI and cell injury by triggering miR-155-HIF-1 alpha pathway, leading to upregulation of HO-1.

7. REFERENCES

1. E. Abraham: Neutrophils and acute lung injury. Crit Care Med, 31(4 Suppl), S195-9 (2003)
DOI: 10.1097/01.CCM.0000057843.47705.E8

2. L. B. Ware and M. A. Matthay: The acute respiratory distress syndrome. N Engl J Med, 342(18), 1334-49 (2000)
DOI: 10.1056/NEJM200005043421806

3. V. Dengler, G. P. Downey, R. M. Tuder, H. K. Eltzschig and E. P. Schmidt: Neutrophil intercellular communication in acute lung injury. Emerging roles of microparticles and gap junctions. Am J Respir Cell Mol Biol, 49(1), 1-5 (2013)
DOI: 10.1165/rcmb.2012-0472TR

4. H. K. Eltzschig and P. Carmeliet: Hypoxia and inflammation. N Engl J Med, 364(7), 656-65 (2011)
DOI: 10.1056/NEJMra0910283

5. W. G. Kaelin, Jr. and P. J. Ratcliffe: Oxygen sensing by metazoans: the central role of the HIF hydroxylase pathway. Mol Cell, 30(4), 393-402 (2008)
DOI: 10.1016/j.molcel.2008.04.009

6. G. L. Semenza: Hydroxylation of HIF-1: oxygen sensing at the molecular level. Physiology (Bethesda), 19, 176-82 (2004)
DOI: 10.1152/physiol.00001.2004

7. J. Hanze, B. G. Eul, R. Savai, S. Krick, P. Goyal, F. Grimminger, W. Seeger and F. Rose: RNA interference for HIF-1alpha inhibits its downstream signalling and affects cellular proliferation. Biochem Biophys Res Commun, 312(3), 571-7 (2003)
DOI: 10.1016/j.bbrc.2003.10.153

8. J. A. Neubauer and J. Sunderram: Heme oxygenase-1 and chronic hypoxia. Respir Physiol Neurobiol, 184(2), 178-85 (2012)
DOI: 10.1016/j.resp.2012.06.027

9. A. Paine, B. Eiz-Vesper, R. Blasczyk and S. Immenschuh: Signaling to heme oxygenase-1 and its anti-inflammatory therapeutic potential. Biochem Pharmacol, 80(12), 1895-903 (2010)
DOI: 10.1016/j.bcp.2010.07.014

10. C. C. Blouin, E. L. Page, G. M. Soucy and D. E. Richard: Hypoxic gene activation by lipopolysaccharide in macrophages: implication of hypoxia-inducible factor 1alpha. Blood, 103(3), 1124-30 (2004)
DOI: 10.1182/blood-2003-07-2427

11. H. J. Knowles, D. R. Mole, P. J. Ratcliffe and A. L. Harris: Normoxic stabilization of hypoxia-inducible factor-1alpha by modulation of the labile iron pool in differentiating U937 macrophages: effect of natural resistance-associated macrophage protein 1. Cancer Res, 66(5), 2600-7 (2006)
DOI: 66/5/2600

12. A. Kuschel, P. Simon and S. Tug: Functional regulation of HIF-1alpha under normoxia--is there more than post-translational regulation? J Cell Physiol, 227(2), 514-24 (2012)
DOI: 10.1002/jcp.22798

13. J. Westra, E. Brouwer, I. A. van Roosmalen, B. Doornbos-van der Meer, M. A. van Leeuwen, M. D. Posthumus and C. G. Kallenberg: Expression and regulation of HIF-1alpha in macrophages under inflammatory conditions; significant reduction of VEGF by CaMKII inhibitor. BMC Musculoskelet Disord, 11, 61 (2010)
DOI: 10.1186/1471-2474-11-61

14. D. P. Bartel: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 116(2), 281-97 (2004)
DOI: S0092867404000455

15. H. Guo, N. T. Ingolia, J. S. Weissman and D. P. Bartel: Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature, 466(7308), 835-40 (2010)
DOI: nature09267

16. R. M. O’Connell, K. D. Taganov, M. P. Boldin, G. Cheng and D. Baltimore: MicroRNA-155 is induced during the macrophage inflammatory response. Proc Natl Acad Sci U S A, 104(5), 1604-9 (2007)
DOI: 0610731104

17. S. T. Cheung, E. Y. So, D. Chang, A. Ming-Lum and A. L. Mui: Interleukin-10 inhibits lipopolysaccharide induced miR-155 precursor stability and maturation. PLoS One, 8(8), e71336 (2013)
DOI: 10.1371/journal.pone.0071336

18. E. Tili, J. J. Michaille, A. Cimino, S. Costinean, C. D. Dumitru, B. Adair, M. Fabbri, H. Alder, C. G. Liu, G. A. Calin and C. M. Croce: Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-alpha stimulation and their possible roles in regulating the response to endotoxin shock. J Immunol, 179(8), 5082-9 (2007)
DOI: 179/8/5082

19. U. Bruning, L. Cerone, Z. Neufeld, S. F. Fitzpatrick, A. Cheong, C. C. Scholz, D. A. Simpson, M. O. Leonard, M. M. Tambuwala, E. P. Cummins and C. T. Taylor: MicroRNA-155 promotes resolution of hypoxia-inducible factor 1alpha activity during prolonged hypoxia. Mol Cell Biol, 31(19), 4087-96 (2011)
DOI: MCB.01276-10

20. M. E. Ostermann, S. P. Keenan, R. A. Seiferling and W. J. Sibbald: Sedation in the intensive care unit: a systematic review. JAMA, 283(11), 1451-9 (2000)
DOI: jce80003

21. M. Flondor, C. Hofstetter, K. A. Boost, C. Betz, M. Homann and B. Zwissler: Isoflurane inhalation after induction of endotoxemia in rats attenuates the systemic cytokine response. Eur Surg Res, 40(1), 1-6 (2008)
DOI: 000107614

22. Q. F. Li, Y. S. Zhu, H. Jiang, H. Xu and Y. Sun: Heme oxygenase-1 mediates the anti-inflammatory effect of isoflurane preconditioning in LPS-stimulated macrophages. Acta Pharmacol Sin, 30(2), 228-34 (2009)
DOI: aps200819

23. Q. F. Li, Y. S. Zhu, H. Jiang, H. Xu and Y. Sun: Isoflurane preconditioning ameliorates endotoxin-induced acute lung injury and mortality in rats. Anesth Analg, 109(5), 1591-7 (2009)
DOI: 109/5/1591

24. X. Dong, R. Hu, Y. Sun, Q. Li and H. Jiang: Isoflurane post-treatment improves pulmonary vascular permeability via upregulation of heme oxygenase-1. Exp Lung Res, 39(7), 295-303 (2013)
DOI: 10.3109/01902148.2013.817627

25. D. N. Carney, P. A. Bunn, Jr., A. F. Gazdar, J. A. Pagan and J. D. Minna: Selective growth in serum-free hormone-supplemented medium of tumor cells obtained by biopsy from patients with small cell carcinoma of the lung. Proc Natl Acad Sci U S A, 78(5), 3185-9 (1981)
DOI: 10.1016/PNAS1981

26. H. J. Kim, J. S. Jeong, S. R. Kim, S. Y. Park, H. J. Chae and Y. C. Lee: Inhibition of endoplasmic reticulum stress alleviates lipopolysaccharide-induced lung inflammation through modulation of NF-kappaB/HIF-1alpha signaling pathway. Sci Rep, 3, 1142 (2013)
DOI: 10.1038/srep01142

27. H. Jiang, Y. Huang, H. Xu, Y. Sun, N. Han and Q. F. Li: Hypoxia inducible factor-1alpha is involved in the neurodegeneration induced by isoflurane in the brain of neonatal rats. J Neurochem, 120(3), 453-60 (2012)
DOI: 10.1111/j.1471-4159.2011.07589.x

28. Y. Sun, Q. F. Li, Y. Zhang, R. Hu and H. Jiang: Isoflurane preconditioning increases survival of rat skin random-pattern flaps by induction of HIF-1alpha expression. Cell Physiol Biochem, 31(4-5), 579-91 (2013)
DOI: 000350078

29. J. M. Sonner, D. Gong, J. Li, E. I. Eger, 2nd and M. J. Laster: Mouse strain modestly influences minimum alveolar anesthetic concentration and convulsivity of inhaled compounds. Anesth Analg, 89(4), 1030-4 (1999)
DOI: 10.3109/1999-1030-4

30. M. A. Croyle, S. M. Callahan, A. Auricchio, G. Schumer, K. D. Linse, J. M. Wilson, L. J. Brunner and G. P. Kobinger: PEGylation of a vesicular stomatitis virus G pseudotyped lentivirus vector prevents inactivation in serum. J Virol, 78(2), 912-21 (2004)
DOI: 10.1128/JVI.78.2.912–921.2004

31. R. Y. Ding, D. M. Zhao, Z. D. Zhang, R. X. Guo and X. C. Ma: Pretreatment of Rho kinase inhibitor inhibits systemic inflammation and prevents endotoxin-induced acute lung injury in mice. J Surg Res, 171(2), e209-14 (2011)
DOI: S0022-4804(11)00723-2

32. G. Matute-Bello, R. K. Winn, M. Jonas, E. Y. Chi, T. R. Martin and W. C. Liles: Fas (CD95) induces alveolar epithelial cell apoptosis in vivo: implications for acute pulmonary inflammation. Am J Pathol, 158(1), 153-61 (2001)
DOI: S0002-9440(10)63953-3

33. L. Cao, C. Feng, L. Li and Z. Zuo: Contribution of microRNA-203 to the isoflurane preconditioning-induced neuroprotection. Brain Res Bull, 88(5), 525-8 (2012)
DOI: S0361-9230(12)00101-3

34. I. S. Chung, J. A. Kim, H. S. Choi, J. J. Lee, M. Yang, H. J. Ahn and S. M. Lee: Reactive oxygen species by isoflurane mediates inhibition of nuclear factor kappaB activation in lipopolysaccharide-induced acute inflammation of the lung. Anesth Analg, 116(2), 327-35 (2013)
DOI: ANE.0b013e31827aec06

35. Y. L. Pang, B. S. Chen, S. P. Li, C. C. Huang, S. W. Chang, C. F. Lam and Y. C. Tsai: The preconditioning pulmonary protective effect of volatile isoflurane in acute lung injury is mediated by activation of endogenous iNOS. J Anesth, 26(6), 822-8 (2012)
DOI: 10.1007/s00540-012-1456-9

36. J. Reutershan, D. Chang, J. K. Hayes and K. Ley: Protective effects of isoflurane pretreatment in endotoxin-induced lung injury. Anesthesiology, 104(3), 511-7 (2006)
DOI: 00000542-200603000-00019

37. X. Xu, J. A. Kim and Z. Zuo: Isoflurane preconditioning reduces mouse microglial activation and injury induced by lipopolysaccharide and interferon-gamma. Neuroscience, 154(3), 1002-8 (2008)
DOI: S0306-4522(08)00540-X

38. Y. Saini, K. K. Greenwood, C. Merrill, K. Y. Kim, S. Patial, N. Parameswaran, J. R. Harkema and J. J. LaPres: Acute cobalt-induced lung injury and the role of hypoxia-inducible factor 1alpha in modulating inflammation. Toxicol Sci, 116(2), 673-81 (2010)
DOI: kfq155

39. Y. Saini, K. Y. Kim, R. Lewandowski, L. A. Bramble, J. R. Harkema and J. J. Lapres: Role of hypoxia-inducible factor 1{alpha} in modulating cobalt-induced lung inflammation. Am J Physiol Lung Cell Mol Physiol, 298(2), L139-47 (2010)
DOI: 00252.2009

40. J. W. Zmijewski, E. Lorne, X. Zhao, Y. Tsuruta, Y. Sha, G. Liu, G. P. Siegal and E. Abraham: Mitochondrial respiratory complex I regulates neutrophil activation and severity of lung injury. Am J Respir Crit Care Med, 178(2), 168-79 (2008)
DOI: 200710-1602OC

41. N. Koshikawa, J. Hayashi, A. Nakagawara and K. Takenaga: Reactive oxygen species-generating mitochondrial DNA mutation up-regulates hypoxia-inducible factor-1alpha gene transcription via phosphatidylinositol 3-kinase-Akt/protein kinase C/histone deacetylase pathway. J Biol Chem, 284(48), 33185-94 (2009)
DOI: M109.054221

42. B. Y. Chin, G. Jiang, B. Wegiel, H. J. Wang, T. Macdonald, X. C. Zhang, D. Gallo, E. Cszimadia, F. H. Bach, P. J. Lee and L. E. Otterbein: Hypoxia-inducible factor 1alpha stabilization by carbon monoxide results in cytoprotective preconditioning. Proc Natl Acad Sci U S A, 104(12), 5109-14 (2007)
DOI: 0609611104

43. T. Cramer, Y. Yamanishi, B. E. Clausen, I. Forster, R. Pawlinski, N. Mackman, V. H. Haase, R. Jaenisch, M. Corr, V. Nizet, G. S. Firestein, H. P. Gerber, N. Ferrara and R. S. Johnson: HIF-1alpha is essential for myeloid cell-mediated inflammation. Cell, 112(5), 645-57 (2003)
DOI: S0092867403001545

44. G. Wang, D. Han, Y. Zhang, X. Xie, Y. Wu, S. Li and M. Li: A novel hypothesis: up-regulation of HO-1 by activation of PPARgamma inhibits HMGB1-RAGE signaling pathway and ameliorates the development of ALI/ARDS. J Thorac Dis, 5(5), 706-10 (2013)
DOI: 10.3978/j.issn.2072-1439.2013.08.69

45. J. Athale, A. Ulrich, N. Chou Macgarvey, R. R. Bartz, K. E. Welty-Wolf, H. B. Suliman and C. A. Piantadosi: Nrf2 promotes alveolar mitochondrial biogenesis and resolution of lung injury in Staphylococcus aureus pneumonia in mice. Free Radic Biol Med, 53(8), 1584-94 (2012)
DOI: S0891-5849(12)00498-4

46. A. Yoshimura, T. Naka and M. Kubo: SOCS proteins, cytokine signalling and immune regulation. Nat Rev Immunol, 7(6), 454-65 (2007)
DOI: nri2093

47. M. J. Rauh, L. M. Sly, J. Kalesnikoff, M. R. Hughes, L. P. Cao, V. Lam and G. Krystal: The role of SHIP1 in macrophage programming and activation. Biochem Soc Trans, 32(Pt 5), 785-8 (2004)
DOI: BST0320785

48. Y. Chen, W. Liu, T. Sun, Y. Huang, Y. Wang, D. K. Deb, D. Yoon, J. Kong, R. Thadhani and Y. C. Li: 1,25-Dihydroxyvitamin D promotes negative feedback regulation of TLR signaling via targeting microRNA-155-SOCS1 in macrophages. J Immunol, 190(7), 3687-95 (2013)
DOI: jimmunol.1203273

49. R. M. O’Connell, A. A. Chaudhuri, D. S. Rao and D. Baltimore: Inositol phosphatase SHIP1 is a primary target of miR-155. Proc Natl Acad Sci U S A, 106(17), 7113-8 (2009)
DOI: 0902636106

Abbreviations: ALI, acute lung injury; LPS, lipopolysaccharide; HIF-1α, Hypoxia inducible factor 1 alpha; HO-1, heme oxygenase-1; miRNAs, microRNAs; UTRs, untranslated regions; TLR, toll like receptor; H&E, hematoxylin and eosin; BALF, bronchoalveolar lavage fluid; CLP, cecal ligation and puncture

Key Words: Isoflurane, Posttreatment, Endotoxin, Acute Lung Injury, miR-155, HIF1 alpha

Send correspondence to: Hong Jiang. Department of Anesthesiology, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, No. 639 Zhi Zao Ju Road, Shanghai, China, Tel: 86-21-23271699, Fax: 86-21-23271699, E-mail: dr_jianghong@163.com