title: Hepatocyte growth factor inhibits hypoxia/reoxygenation-induced activation of xanthine oxidase in endothelial cells through the JAK2 signaling pathway.
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+.
authors: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors: Ying Qian
authors: YQ
authors: Zhang
authors: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors: Shun Ying
authors: SY
authors: Hu
authors: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors: Yun Dai
authors: YD
authors: Chen
authors: Department of Gastroenterology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors: Ming Zhou
authors: MZ
authors: Guo
authors: Central Laboratory, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors: Shan
authors: S
authors: Wang
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D015687:Cell Hypoxia / Q000187:drug effects; D003600:Cytosol / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D053614:Janus Kinase 2 / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D014969:Xanthine Oxidase / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D017228:Hepatocyte Growth Factor; D014969:Xanthine Oxidase; D019869:Phosphatidylinositol 3-Kinases; D053614:Janus Kinase 2; D010100:Oxygen; D002118:Calcium