[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


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


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.


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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