Vaya Jacob, Song Wei, Khatib Soliman, Geng Guoyan, Schipper Hyman M
Laboratory of Natural Medicinal Compounds, Migal-Galilee Technological Center, Kiryat-Shmona 11016, Israel.
Free Radic Biol Med. 2007 Mar 15;42(6):864-71. doi: 10.1016/j.freeradbiomed.2006.12.022. Epub 2006 Dec 22.
Up-regulation of heme oxygenase-1 (HO-1) and altered cholesterol metabolism are characteristic of Alzheimer-diseased (AD) neural tissues. Central oxidation of cholesterol to oxysterols has been implicated in neuroembryogenesis, synaptic plasticity, and membrane repair. In the current study, we demonstrated that transient transfection of rat astroglia with human (h)ho-1 cDNA for 3 days significantly decreased intracellular cholesterol concentrations and increased levels of four oxysterol species (measured by GC/MS) compared to untreated control cultures and HO-1-transfected cells exposed to the HO inhibitor, tin mesoporphyrin (SnMP). Relative to control preparations, oxidative stress was augmented in mitochondria (isolated by subcellular fractionation) and culture media derived from HO-1-transfected astrocytes, as evidenced by enhanced oxidation of the synthetic reporter molecules, linoleoyl tyrosine (LT), linoleoyl tyrosine cholesterol ester (LTC), or linoleoyl tyrosine deoxyguanosyl ester (LTG; measured by GC/MS and LC/MS/MS). We also observed enhanced oxidation of exogenous LTC in human neuroblastoma (M17) cells exposed for 18 h to conditioned media collected from HO-1-transfected astrocytes relative to control media. In AD and other pathological states, glial HO-1 induction may transduce ambient noxious stimuli (e.g., beta-amyloid) into altered patterns of glial sterol metabolism which, in turn, may affect neuronal membrane turnover, survival, and adaptability.
血红素加氧酶-1(HO-1)的上调和胆固醇代谢改变是阿尔茨海默病(AD)神经组织的特征。胆固醇在中枢氧化为氧甾醇与神经胚胎发育、突触可塑性和膜修复有关。在本研究中,我们证明,与未处理的对照培养物以及暴露于HO抑制剂中卟啉锡(SnMP)的HO-1转染细胞相比,用人类(h)ho-1 cDNA瞬时转染大鼠星形胶质细胞3天可显著降低细胞内胆固醇浓度,并增加四种氧甾醇种类的水平(通过气相色谱/质谱法测量)。相对于对照制剂,在从HO-1转染的星形胶质细胞分离的线粒体(通过亚细胞分级分离)和培养基中氧化应激增强,这可通过合成报告分子亚油酰酪氨酸(LT)、亚油酰酪氨酸胆固醇酯(LTC)或亚油酰酪氨酸脱氧鸟苷酯(LTG)的氧化增强来证明(通过气相色谱/质谱法和液相色谱/串联质谱法测量)。我们还观察到,相对于对照培养基,暴露于从HO-1转染的星形胶质细胞收集的条件培养基18小时的人神经母细胞瘤(M17)细胞中外源LTC的氧化增强。在AD和其他病理状态下,胶质细胞HO-1的诱导可能将周围的有害刺激(例如β-淀粉样蛋白)转化为胶质细胞甾醇代谢的改变模式,这反过来可能影响神经元膜更新、存活和适应性。