Yan Wang1, Xiao-Yu Jiang, Li Liu, Hui-Qing Jiang. 1. Department of Gastroenterology, The Second Hospital of Hebei Medical University, The Hebei Institute of Gastroenterology, No. 215, Heping West Road, Shijiazhuang 050000, Hebei Province, China.
Abstract
AIM: To investigate the role of phosphatidylinositol 3-kinase (PI 3-K)/Akt signaling pathway in the balance of HSC activation and apoptosis in rat hepatic stellate cells (HSC). METHODS: An activated HSC cell line was used in this study. LY 294002, the PI 3-K/Akt signal pathway blocker was used to investigate the molecular events on apoptosis in HSC and to interpret the role of this pathway in HSC apoptosis. Immunocytochemistry, Western blot and reverse transcription polymerase chain reaction (RT-PCR) analysis were applied to detect the expression of PI 3-K, and simultaneously phosphorylated-Akt (p-Akt) and total-Akt were determined by Western blot. The HSC apoptosis was examined by annexin-V/propidium iodide double-labelled flow cytometry and transmission electron microscopy. RESULTS: The apoptosis rates in LY 294002 (30.82%+/-2.90%) and LY 294002+PDGF-BB (28.16%+/-2.58%) groups were significantly increased compared with those of control (9.02%+/-1.81%) and PDGF-BB (4.35%+/-1.18%). PDGF-BB augmented PI 3-K and p-Akt expression. LY 294002 significantly reduced the contents of PI 3-K and p-Akt. mRNA transcription evaluated by RT-PCR showed similar tendencies as protein expression. CONCLUSION: Inhibition of PI 3-K/Akt signaling pathway induces apoptosis in HSC.
AIM: To investigate the role of phosphatidylinositol 3-kinase (PI 3-K)/Akt signaling pathway in the balance of HSC activation and apoptosis in rat hepatic stellate cells (HSC). METHODS: An activated HSC cell line was used in this study. LY 294002, the PI 3-K/Akt signal pathway blocker was used to investigate the molecular events on apoptosis in HSC and to interpret the role of this pathway in HSC apoptosis. Immunocytochemistry, Western blot and reverse transcription polymerase chain reaction (RT-PCR) analysis were applied to detect the expression of PI 3-K, and simultaneously phosphorylated-Akt (p-Akt) and total-Akt were determined by Western blot. The HSC apoptosis was examined by annexin-V/propidium iodide double-labelled flow cytometry and transmission electron microscopy. RESULTS: The apoptosis rates in LY 294002 (30.82%+/-2.90%) and LY 294002+PDGF-BB (28.16%+/-2.58%) groups were significantly increased compared with those of control (9.02%+/-1.81%) and PDGF-BB (4.35%+/-1.18%). PDGF-BB augmented PI 3-K and p-Akt expression. LY 294002 significantly reduced the contents of PI 3-K and p-Akt. mRNA transcription evaluated by RT-PCR showed similar tendencies as protein expression. CONCLUSION: Inhibition of PI 3-K/Akt signaling pathway induces apoptosis in HSC.
Authors: Frank R Murphy; Razao Issa; Xiaoying Zhou; Shabna Ratnarajah; Hideaki Nagase; Michael J P Arthur; Christopher Benyon; John P Iredale Journal: J Biol Chem Date: 2002-01-16 Impact factor: 5.157
Authors: G Robino; M Parola; F Marra; A Caligiuri; R M De Franco; E Zamara; G Bellomo; P Gentilini; M Pinzani; M U Dianzani Journal: J Biol Chem Date: 2000-12-22 Impact factor: 5.157