Inhibition of the expression of aquaporin‑1 by RNA interference in pulmonary epithelial cells and its effects on water transport

Zhang,Qiuyue; Fu,Jianhua; Xue,Xindong

doi:10.3892/mmr.2015.4519

Inhibition of the expression of aquaporin‑1 by RNA interference in pulmonary epithelial cells and its effects on water transport

Authors:
- Qiuyue Zhang
- Jianhua Fu
- Xindong Xue
View Affiliations

Affiliations: Department of Pediatrics, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Pediatrics, China Medical University Affiliated to Shengjing Hospital, Shenyang, Liaoning 110004, P.R. China
Published online on: November 6, 2015 https://doi.org/10.3892/mmr.2015.4519
Pages: 281-286

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

In the present study, the effect of aquaporin‑1 (AQP1) on fluid transportation in pulmonary epithelial cells, and the role of AQP1 in alveolar fluid clearance were investigated to provide an experimental foundation to elucidate the pathogenesis of hyperoxic lung edema. An siRNA transfection technique was used to silence AQP1 in the A549 cell line. The transfected cells were randomized into a hyperoxia exposure and an air control group, with a negative control group set for each group. Cell volume was determined using flow cytometry, and Pf values were used to determine osmotic water permeability. Cell volume was found to be reduced in the AQP1‑silenced A549 cells, compared with the negative control group 72 h following air exposure. In addition, cell volume was reduced in the AQP1‑silenced A549 cells, compared with the negative control group 48 and 72 h following hyperoxia exposure. The osmotic water permeability of the AQP1‑silenced cells was reduced in the air control and hyperoxia exposure groups, compared with the negative control group 48 and 72 h following exposure. The volume and cell membrane osmotic water permeability of the A549 cells were reduced, compared with those in the control group following AQP1‑silencing, which indicated that the downregulation of AQP1 impedes extracellular to intracellular fluid transportation. Therefore, the disturbance in alveolar fluid clearance resulting from the downregulation of AQP1 following hyperoxia exposure may be one of the key mechanisms responsible for hyperoxic lung edema.

View Figures

View References

1	Chang LW: Part of China's urban premature incidence of bronchial pulmonary dysplasia and risk factors. In: The 9th national neonatal meeting; 2009
2	Qi QY, Chen W, Li XL, Wang YW and Xie XH: H₂S protecting against lung injury following limb ischemia-reperfusion by alleviating inflammation and water transport abnormality in rats. Biomed Environ Sci. 27:410–418. 2014.PubMed/NCBI
3	Elias AS, Oliveira GP, Ornellas DS, Morales MM, Capelozzi VL, Haddad R, Pelosi P, Rocco PR and Garcia CS: Effects of early and late pneumothorax drainage on the development of pulmonary oedema. Respir Physiol Neurobiol. 195:27–36. 2014. View Article : Google Scholar : PubMed/NCBI
4	Wang Q, Ishikawa T, Michiue T, Zhu BL, Guan DW and Maeda H: Molecular pathology of pulmonary edema after injury in forensic autopsy cases. Int J Legal Med. 126:875–882. 2012. View Article : Google Scholar : PubMed/NCBI
5	Zhang QY, Fu JH and Xue XD: Expression and function of aquaporin-1 in hyperoxia-exposed alveolar epithelial type II cells. Exp Ther Med. 8:493–498. 2014.PubMed/NCBI
6	Lecuona E, Trejo HE and Sznajder JI: Regulation of Na, K-ATPase during acute lung injury. J Bioenerg Biomembr. 39:391–395. 2007. View Article : Google Scholar : PubMed/NCBI
7	Winer J, Jung CK, Shackel I and Williams PM: Development and validation of real-time quantitative reverse transcriptase- polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem. 270:41–49. 1999. View Article : Google Scholar : PubMed/NCBI
8	Jordan CT, Yamasaki G and Minamoto D: High-resolution cell cycle analysis of defined phenotypic subsets within primitive human hematopoietic cell populations. Exp Hematol. 24:1347–1355. 1996.PubMed/NCBI
9	Preston GM, Carroll TP, Guggino WB and Agre P: Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science. 256:385–387. 1992. View Article : Google Scholar : PubMed/NCBI
10	Verkman AS: Physiological importance of aquaporin water channels. Ann Med. 34:192–200. 2002. View Article : Google Scholar : PubMed/NCBI
11	Itoh T, Rai T, Kuwahara M, Ko SB, Uchida S, Sasaki S and Ishibashi K: Identification of a novel aquaporin, AQP12, expressed in pancreatic acinar cells. Biochem Biophys Res Commun. 330:832–838. 2005. View Article : Google Scholar : PubMed/NCBI
12	Borok Z and Verkman AS: Lung edema clearance: 20 years of progress: Invited review: Role of aquaporin water channls in fluid transport in lung and airways. J Appl Physiol (1985). 93:2199–2206. 2002. View Article : Google Scholar
13	Jiao GY, Li ER and Yu R: Decreased expression of AQP1 and AQP5 in acute injured lungs in rats. Chin Med J (Engl). 155:963–967. 2002.
14	Towne JE, Harrod KS, Krane CM and Menon AG: Decreased expression of aquaporin (AQP)1 and AQP5 in mouse lung after acute viral infection. Am J Respir Cell Mol Biol. 22:34–44. 2000. View Article : Google Scholar
15	Matthay MA, Folkesson HG and Clerici C: Lung epithelial fluid transport and the resolution of pulmonary edema. Physiol Rev. 82:569–600. 2002. View Article : Google Scholar : PubMed/NCBI
16	Krane CM, Deng B, Mutyam V, McDonald CA, Pazdziorko S, Mason L, Goldman S, Kasaian M, Chaudhary D, Williams C and Ho MW: Altered regulation of aquaporin gene expression in allergen and IL-13-induced mouse models of asthma. Cytokine. 46:111–118. 2009. View Article : Google Scholar : PubMed/NCBI
17	Wang F, Huang H, Lu F and Chen Y: Acute lung injury and change in expression of aquaporins 1 and 5 in a rat model of acute pancreatitis. Hepatogastroenterology. 57:1553–1562. 2010.
18	Wu XM, Wang HY, Li GF, Zang B and Chen WM: Dobutamine enhances alveolar fluid clearance in a rat model of acute lung injury. Lung. 187:225–231. 2009. View Article : Google Scholar : PubMed/NCBI
19	Jin LD, Wang LR, Wu LQ, Shan YL, Zhao XY, Xiong XQ, Zhou JH, Lin LN and Jin LL: Effects of COX-2 inhibitor on ventilator-induced lung injury in rats. Int Immunopharmacol. 16:288–295. 2013. View Article : Google Scholar : PubMed/NCBI
20	Gao Z, Xu J, Sun D, Zhang R, Liang R, Wang L and Fan R: Traditional Chinese medicine, Qing Ying Tang, ameliorates the severity of acute lung injury induced by severe acute pancreatitis in rats via the upregulation of aquaporin-1. Exp Ther Med. 8:1819–1824. 2014.PubMed/NCBI
21	Song Y, Ma T, Mathay MA and Verkman AS: Role of aquaporin-4 in airspace-to-capillary water permeability in intact mouse lung measured by a novel gravimetric method. J Gen Physol. 155:17–27. 2000.
22	King LS, Nielsen S, Agre P and Brown RH: Decreased pulmonary vascular permeability in aquaporin-1-null humans. Proc Natl Acad Sci USA. 99:1059–1063. 2002. View Article : Google Scholar : PubMed/NCBI
23	Li J, Xu M, Fan Q, Xie X, Zhang Y, Mu D, Zhao P, Zhang B, Cao F, Wang Y, et al: Tanshinone IIA ameliorates seawater exposure-induced lung injury by inhibiting aquaporins (AQP) 1 and AQP5 expression in lung. Respir Physiol Neurobiol. 176:39–49. 2011. View Article : Google Scholar : PubMed/NCBI
24	Lai KN, Leung JC, Metz CN, Lai FM, Bucala R and Lan HY: Role for macrophage migration inhibitory factor in acute respiratory distress syndrome. J Pathol. 199:496–508. 2003. View Article : Google Scholar : PubMed/NCBI
25	Song Y, Fukuda N, Bai C, Ma T, Matthay MA and Verkman AS: Role of aquaporins in alveolar fluid clearance in neonatal and adult lung and in oedema formation following acute lung injury: Studies in transgenic aquaporin null mice. J Physiol. 525:771–779. 2000. View Article : Google Scholar
26	Modi N: Clinical implications of postnatal alterations in body water distribution. Semin Neonatol. 8:301–306. 2003. View Article : Google Scholar
27	Chen F, Zhang C, Jia X, Wang S, Wang J, Chen Y, Zhao J, Tian S, Han X and Han L: Transcriptome profiles of human lung epithelial cells A549 interacting with aspergillus fumigatus by RNA-Seq. PLoS One. 10:e01357202015. View Article : Google Scholar : PubMed/NCBI
28	Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE and Mello CC: Potent and spectific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391:806–811. 1998. View Article : Google Scholar : PubMed/NCBI
29	Aagaard L and Rossi JJ: RNAi therapeutics: Principles, prospects and chanllenges. Adv Drug Deliv Rev. 59:75–86. 2007. View Article : Google Scholar : PubMed/NCBI