Du Botao, Ohmichi Masahide, Takahashi Kazuhiro, Kawagoe Jun, Ohshima Chika, Igarashi Hideki, Mori-Abe Akiko, Saitoh Maki, Ohta Tsuyoshi, Ohishi Akira, Doshida Masakazu, Tezuka Naohiro, Takahashi Toshifumi, Kurachi Hirohisa
Department of Obstetrics and Gynecology, Yamagata University, School of Medicine, 2-2-2 Iidanishi, Yamagata 990-9585, Japan.
J Endocrinol. 2004 Dec;183(3):605-15. doi: 10.1677/joe.1.05775.
Although estrogen is known to protect against beta-amyloid (Abeta)-induced neurotoxicity, the mechanisms responsible for this effect are only beginning to be elucidated. In addition, the effect of raloxifene on Abeta-induced neuro-toxicity remains unknown. Here we investigated whether raloxifene exhibits similar neuro-protective effects to estrogen against Abeta-induced neurotoxicity and the mechanism of the effects of these agents in PC12 cells transfected with the full-length human estrogen receptor (ER) alpha gene (PCER). Raloxifene, like 17beta-estradiol (E2), significantly inhibited Abeta-induced apoptosis in PCER cells, but not in a control line of cells transfected with vector DNA alone (PCCON). Since telomerase activity, the level of which is modulated by regulation of telomerase catalytic subunit (TERT) at both the transcriptional and post-transcriptional levels, is known to be involved in suppressing apoptosis in neurons, we examined the effect of E2 and raloxifene on telomerase activity. Although both E2 and raloxifene induced telomerase activity in PCER cells, but not in PCCON cells, treated with Abeta, they had no effect on the level of TERT expression. These results suggest that neither E2 nor raloxifene affects the telomerase activity at the transcriptional level. We therefore studied the mechanism by which E2 and raloxifene induce the telomerase activity at the post-transcriptional level. Both E2 and raloxifene induced the phosphorylation of Akt, and pre-treatment with a phosphatidylinositol 3-kinase inhibitor, LY294002, attenuated both E2- and raloxifene-induced activation of the telomerase activity. Moreover, both E2 and raloxifene induced both the phosphorylation of TERT at a putative Akt phosphorylation site and the association of nuclear factor kappaB with TERT. Our findings suggest that and raloxifene exert neuroprotective effects by E2 telomerase activation via a post-transcriptional cascade in an experimental model relevant to Alzheimer's disease.
尽管已知雌激素可预防β-淀粉样蛋白(Aβ)诱导的神经毒性,但其作用机制才刚刚开始被阐明。此外,雷洛昔芬对Aβ诱导的神经毒性的影响尚不清楚。在此,我们研究了雷洛昔芬是否对Aβ诱导的神经毒性表现出与雌激素相似的神经保护作用,以及这些药物在转染了全长人雌激素受体(ER)α基因的PC12细胞(PCER)中的作用机制。雷洛昔芬与17β-雌二醇(E2)一样,显著抑制PCER细胞中Aβ诱导的细胞凋亡,但对仅转染载体DNA的对照细胞系(PCCON)无此作用。由于端粒酶活性在转录和转录后水平均受端粒酶催化亚基(TERT)调控,且已知其参与抑制神经元凋亡,因此我们检测了E2和雷洛昔芬对端粒酶活性的影响。尽管E2和雷洛昔芬均可在Aβ处理的PCER细胞中诱导端粒酶活性,但对PCCON细胞无此作用,且它们对TERT表达水平无影响。这些结果表明,E2和雷洛昔芬均未在转录水平影响端粒酶活性。因此,我们研究了E2和雷洛昔芬在转录后水平诱导端粒酶活性的机制。E2和雷洛昔芬均可诱导Akt磷酸化,用磷脂酰肌醇3-激酶抑制剂LY294002预处理可减弱E2和雷洛昔芬诱导的端粒酶活性激活。此外,E2和雷洛昔芬均可诱导TERT在假定的Akt磷酸化位点磷酸化以及核因子κB与TERT结合。我们的研究结果表明,在与阿尔茨海默病相关的实验模型中,E2和雷洛昔芬通过转录后级联反应激活端粒酶发挥神经保护作用。