Wang Yan, Jiang Xiao-Yu, Liu Li, Jiang Hui-Qing
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.
World J Gastroenterol. 2008 Sep 7;14(33):5186-91. doi: 10.3748/wjg.14.5186.
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).
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.
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.
Inhibition of PI 3-K/Akt signaling pathway induces apoptosis in HSC.
探讨磷脂酰肌醇3激酶（PI 3-K）/Akt信号通路在大鼠肝星状细胞（HSC）激活与凋亡平衡中的作用。
本研究采用激活的HSC细胞系。使用PI 3-K/Akt信号通路阻滞剂LY 294002研究HSC凋亡的分子事件，并阐释该通路在HSC凋亡中的作用。应用免疫细胞化学、蛋白质印迹法及逆转录聚合酶链反应（RT-PCR）分析检测PI 3-K的表达，同时通过蛋白质印迹法测定磷酸化Akt（p-Akt）和总Akt。采用膜联蛋白V/碘化丙啶双染流式细胞术和透射电子显微镜检测HSC凋亡情况。
与对照组（9.02%±1.81%）和血小板衍生生长因子BB（PDGF-BB）组（4.35%±1.18%）相比，LY 294002组（30.82%±2.90%）和LY 294002 + PDGF-BB组（28.16%±2.58%）的凋亡率显著升高。PDGF-BB增强了PI 3-K和p-Akt的表达。LY 294002显著降低了PI 3-K和p-Akt的含量。RT-PCR评估的mRNA转录显示出与蛋白表达相似的趋势。
抑制PI 3-K/Akt信号通路可诱导HSC凋亡。
