Hepatocyte growth factor inhibits hypoxia/reoxygenation-induced activation of xanthine oxidase in endothelial cells through the JAK2 signaling pathway

Zhang,Ying  Qian; Hu,Shun  Ying; Chen,Yun  Dai; Guo,Ming  Zhou; Wang,Shan

doi:10.3892/ijmm.2016.2708

Hepatocyte growth factor inhibits hypoxia/reoxygenation-induced activation of xanthine oxidase in endothelial cells through the JAK2 signaling pathway

Authors:
- Ying Qian Zhang
- Shun Ying Hu
- Yun Dai Chen
- Ming Zhou Guo
- Shan Wang
View Affiliations

Affiliations: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China, Department of Gastroenterology, Chinese PLA General Hospital, Beijing 100853, P.R. China, Central Laboratory, Chinese PLA General Hospital, Beijing 100853, P.R. China
Published online on: August 18, 2016 https://doi.org/10.3892/ijmm.2016.2708
Pages: 1055-1062
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Vascular endothelial cells (ECs) appear to be one of the primary targets of hypoxia/reoxygenation (H/R) injury. In our previous study, we demonstrated that hepatocyte growth factor (HGF) exhibited a protective effect in cardiac microvascular endothelial cells (CMECs) subjected to H/R by inhibiting xanthine oxidase (XO) by reducing the cytosolic Ca2+ concentration increased in response to H/R. The precise mechanisms through which HGF inhibits XO activation remain to be determined. In the present study, we examined the signaling pathway through which HGF regulates Ca2+ concentrations and the activation of XO during H/R in primary cultured rat CMECs. CMECs were exposed to 4 h of hypoxia and 1 h of reoxygenation. The protein expression of XO and the activation of the phosphoinositide 3-kinase (PI3K), janus kinase 2 (JAK2) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways were detected by western blot analysis. Cytosolic calcium (Ca2+) concentrations and reactive oxygen species (ROS) levels were measured by flow cytometry. The small interfering RNA (siRNA)‑mediated knockdown of XO inhibited the increase in ROS production induced by H/R. LY294002 and AG490 inhibited the H/R-induced increase in the production and activation of XO. The PI3K and JAK2 signaling pathways were activated by H/R. The siRNA‑mediated knockdown of PI3K and JAK2 also inhibited the increase in the production of XO protein. HGF inhibited JAK2 activation whereas it had no effect on PI3K activation. The siRNA-mediated knockdown of JAK2 prevented the increase in cytosolic Ca2+ induced by H/R. Taken together, these findings suggest that H/R induces the production and activation of XO through the JAK2 and PI3K signaling pathways. Furthermore, HGF prevents XO activation following H/R primarily by inhibiting the JAK2 signaling pathway and in turn, inhibiting the increase in cytosolic Ca2+.

View Figures

View References

1	Yu G, Peng T, Feng Q and Tyml K: Abrupt reoxygenation of microvascular endothelial cells after hypoxia activates ERK1/2 and JNK1, leading to NADPH oxidase-dependent oxidant production. Microcirculation. 14:125–136. 2007. View Article : Google Scholar : PubMed/NCBI
2	Zhang T, Yang D, Fan Y, Xie P and Li H: Epigallocatechin-3-gallate enhances ischemia/reperfusion-induced apoptosis in human umbilical vein endothelial cells via AKT and MAPK pathways. Apoptosis. 14:1245–1254. 2009. View Article : Google Scholar : PubMed/NCBI
3	Dhar-Mascareño M, Cárcamo JM and Golde DW: Hypoxia-reoxygenation-induced mitochondrial damage and apoptosis in human endothelial cells are inhibited by vitamin C. Free Radic Biol Med. 38:1311–1322. 2005. View Article : Google Scholar : PubMed/NCBI
4	Pearlstein DP, Ali MH, Mungai PT, Hynes KL, Gewertz BL and Schumacker PT: Role of mitochondrial oxidant generation in endothelial cell responses to hypoxia. Arterioscler Thromb Vasc Biol. 22:566–573. 2002. View Article : Google Scholar : PubMed/NCBI
5	Jang H-J, Koo BK, Lee HS, Park JB, Kim JH, Seo MK, Yang HM, Park KW, Nam CW, Doh JH and Kim HS: Safety and efficacy of a novel hyperaemic agent, intracoronary nicorandil, for invasive physiological assessments in the cardiac catheterization laboratory. Eur Heart J. 34:2055–2062. 2013. View Article : Google Scholar : PubMed/NCBI
6	Meneshian A and Bulkley GB: The physiology of endothelial xanthine oxidase: from urate catabolism to reperfusion injury to inflammatory signal transduction. Microcirculation. 9:161–175. 2002. View Article : Google Scholar : PubMed/NCBI
7	Landmesser U, Spiekermann S, Preuss C, Sorrentino S, Fischer D, Manes C, Mueller M and Drexler H: Angiotensin II induces endothelial xanthine oxidase activation: role for endothelial dysfunction in patients with coronary disease. Arterioscler Thromb Vasc Biol. 27:943–948. 2007. View Article : Google Scholar : PubMed/NCBI
8	Spiekermann S, Landmesser U, Dikalov S, Bredt M, Gamez G, Tatge H, Reepschläger N, Hornig B, Drexler H and Harrison DG: Electron spin resonance characterization of vascular xanthine and NAD(P)H oxidase activity in patients with coronary artery disease: Relation to endothelium-dependent vasodilation. Circulation. 107:1383–1389. 2003. View Article : Google Scholar : PubMed/NCBI
9	Berry CE and Hare JM: Xanthine oxidoreductase and cardiovascular disease: molecular mechanisms and pathophysiological implications. J Physiol. 555:589–606. 2004. View Article : Google Scholar
10	Sulikowski T, Domanski L, Ciechanowski K, Adler G, Pawlik A, Safranow K, Dziedziejko V, Chlubek D and Ciechanowicz A: Effect of trimetazidine on xanthine oxidoreductase expression in rat kidney with ischemia-reperfusion injury. Arch Med Res. 39:459–462. 2008. View Article : Google Scholar : PubMed/NCBI
11	McNally JS, Saxena A, Cai H, Dikalov S and Harrison DG: Regulation of xanthine oxidoreductase protein expression by hydrogen peroxide and calcium. Arterioscler Thromb Vasc Biol. 25:1623–1628. 2005. View Article : Google Scholar : PubMed/NCBI
12	Kayyali US, Donaldson C, Huang H, Abdelnour R and Hassoun PM: Phosphorylation of xanthine dehydrogenase/oxidase in hypoxia. J Biol Chem. 276:14359–14365. 2001.PubMed/NCBI
13	Wang G, Qian P, Jackson FR, Qian G and Wu G: Sequential activation of JAKs, STATs and xanthine dehydrogenase/oxidase by hypoxia in lung microvascular endothelial cells. Int J Biochem. Cell Biol. 40:461–470. 2008.
14	Reviriego-Mendoza MM and Santy LC: The cytohesin guanosine exchange factors (GEFs) are required to promote HGF-mediated renal recovery after acute kidney injury (AKI) in mice. Physiol Rep. 3:e124422015. View Article : Google Scholar : PubMed/NCBI
15	White HM, Acton AJ, Kamocka MM and Considine RV: Hepatocyte growth factor regulates neovascularization in developing fat pads. Am J Physiol Endocrinol Metab. 306:E189–E196. 2014. View Article : Google Scholar :
16	Zhang Y, Hu S and Chen Y: Hepatocyte growth factor suppresses hypoxia/reoxygenationinduced XO activation in cardiac microvascular endothelial cells. Heart Vessels. 30:534–544. 2015. View Article : Google Scholar
17	Nishida M, Carley WW, Gerritsen ME, Ellingsen O, Kelly RA and Smith TW: Isolation and characterization of human and rat cardiac microvascular endothelial cells. Am J Physiol. 264:H639–H652. 1993.PubMed/NCBI
18	Zhang Z, Li W, Sun D, Zhao L, Zhang R, Wang Y, Zhou X, Wang H and Cao F: Toll-like receptor 4 signaling in dysfunction of cardiac microvascular endothelial cells under hypoxia/reoxygenation. Inflamm Res. 60:37–45. 2011. View Article : Google Scholar
19	Ladilov Y, Schäfer C, Held A, Schäfer M, Noll T and Piper HM: Mechanism of Ca(2+) overload in endothelial cells exposed to simulated ischemia. Cardiovasc Res. 47:394–403. 2000. View Article : Google Scholar : PubMed/NCBI
20	Yu G, Bolon M, Laird DW and Tyml K: Hypoxia and reoxygenation-induced oxidant production increase in microvascular endothelial cells depends on connexin40. Free Radic Biol Med. 49:1008–1013. 2010. View Article : Google Scholar : PubMed/NCBI
21	Loor G, Kondapalli J, Iwase H, Chandel NS, Waypa GB, Guzy RD, Vanden Hoek TL and Schumacker PT: Mitochondrial oxidant stress triggers cell death in simulated ischemia-reperfusion. Biochim Biophys Acta. 1813:1382–1394. 2011. View Article : Google Scholar :
22	Kong R, Jia G, Cheng ZX, Wang YW, Mu M, Wang SJ, Pan SH, Gao Y, Jiang HC, Dong DL and Sun B: Dihydroartemisinin enhances Apo2L/TRAIL-mediated apoptosis in pancreatic cancer cells via ROS-mediated up-regulation of death receptor 5. PLoS One. 7:e372222012. View Article : Google Scholar : PubMed/NCBI
23	Przygodzki T, Sokal A and Bryszewska M: Calcium ionophore A23187 action on cardiac myocytes is accompanied by enhanced production of reactive oxygen species. Biochim Biophys Acta. 1740:481–488. 2005. View Article : Google Scholar : PubMed/NCBI
24	Rasband WS: ImageJ. U.S National Institutes of Health; Bethesda, MD: 1997–2012
25	Chatterjee S, Browning EA, Hong N, DeBolt K, Sorokina EM, Liu W, Birnbaum MJ and Fisher AB: Membrane depolarization is the trigger for PI3K/Akt activation and leads to the generation of ROS. Am J Physiol Heart Circ Physiol. 302:H105–H114. 2012. View Article : Google Scholar :
26	Kelley EE, Khoo NK, Hundley NJ, Malik UZ, Freeman BA and Tarpey MM: Hydrogen peroxide is the major oxidant product of xanthine oxidase. Free Radic Biol Med. 48:493–498. 2010. View Article : Google Scholar :
27	Saito S, Ogawa J and Minamiya Y: Pulmonary reexpansion causes xanthine oxidase-induced apoptosis in rat lung. Am J Physiol Lung Cell Mol Physiol. 289:L400–L406. 2005. View Article : Google Scholar : PubMed/NCBI
28	Krause GS, White BC, Aust SD, Nayini NR and Kumar K: Brain cell death following ischemia and reperfusion: A proposed biochemical sequence. Crit Care Med. 16:714–726. 1988. View Article : Google Scholar : PubMed/NCBI
29	Ono T, Tsuruta R, Fujita M, Aki HS, Kutsuna S, Kawamura Y, Wakatsuki J, Aoki T, Kobayashi C, Kasaoka S, et al: Xanthine oxidase is one of the major sources of superoxide anion radicals in blood after reperfusion in rats with forebrain ischemia/reperfusion. Brain Res. 1305:158–167. 2009. View Article : Google Scholar : PubMed/NCBI
30	Therade-Matharan S, Laemmel E, Duranteau J and Vicaut E: Reoxygenation after hypoxia and glucose depletion causes reactive oxygen species production by mitochondria in HUVEC. Am J Physiol Regul Integr Comp Physiol. 287:R1037–R1043. 2004. View Article : Google Scholar : PubMed/NCBI
31	Dai T, Zheng H and Fu GS: Hypoxia confers protection against apoptosis via the PI3K/Akt pathway in endothelial progenitor cells. Acta Pharmacol Sin. 29:1425–1431. 2008. View Article : Google Scholar : PubMed/NCBI
32	Shimizu S, Yonezawa R, Hagiwara T, Yoshida T, Takahashi N, Hamano S, Negoro T, Toda T, Wakamori M, Mori Y and Ishii M: Inhibitory effects of AG490 on H₂O₂-induced TRPM2-mediated Ca(2+) entry. Eur J Pharmacol. 742:22–30. 2014. View Article : Google Scholar : PubMed/NCBI
33	Saksela M, Lapatto R and Raivio KO: Irreversible conversion of xanthine dehydrogenase into xanthine oxidase by a mitochondrial protease. FEBS Lett. 443:117–120. 1999. View Article : Google Scholar : PubMed/NCBI