Contreras Miguel Agustin, Haq Ehtishamul, Uto Takuhiro, Singh Inderjit, Singh Avtar Kaur
Department of Pediatrics and The Charles Darby Children's Research Institute, Medical University of South Carolina, Charleston, SC, USA.
Arch Biochem Biophys. 2008 Sep 15;477(2):211-8. doi: 10.1016/j.abb.2008.06.012. Epub 2008 Jun 24.
Krabbe disease is a neuroinflammatory disorder in which galactosylsphingosine (psychosine) accumulates in nervous tissue. To gain insight into whether the psychosine-induced effects in nervous tissue extend to peripheral organs, we investigated the expression of cytokines and their effects on peroxisomal structure/functions in twitcher mouse liver (animal model of Krabbe disease). Immunofluorescence analysis demonstrated TNF-alpha and IL-6 expression, which was confirmed by mRNAs quantitation. Despite the presence of TNF-alpha, lipidomic analysis did not indicate a significant decrease in sphingomyelin or an increase in ceramide fractions. Ultrastructural analysis of catalase-dependent staining of liver sections showed reduced reactivity without significant changes in peroxisomal contents. This observation was confirmed by assaying catalase activity and quantitation of its mRNA, both of which were found significantly decreased in twitcher mouse liver. Western blot analysis demonstrated a generalized reduction of peroxisomal matrix and membrane proteins. These observations indicate that twitcher mouse pathobiology extends to the liver, where psychosine-induced TNF-alpha and IL-6 compromise peroxisomal structure and functions.
克拉伯病是一种神经炎症性疾病,其中半乳糖基鞘氨醇(精神鞘氨醇)在神经组织中蓄积。为深入了解精神鞘氨醇在神经组织中诱导的效应是否会扩展到外周器官,我们研究了细胞因子在震颤小鼠肝脏(克拉伯病动物模型)中的表达及其对过氧化物酶体结构/功能的影响。免疫荧光分析证实了肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达,mRNA定量分析也证实了这一点。尽管存在TNF-α,但脂质组学分析并未表明鞘磷脂有显著减少或神经酰胺组分增加。对肝切片进行过氧化氢酶依赖性染色的超微结构分析显示反应性降低,而过氧化物酶体内容物无明显变化。通过检测过氧化氢酶活性及其mRNA定量分析证实了这一观察结果,两者在震颤小鼠肝脏中均显著降低。蛋白质印迹分析表明过氧化物酶体基质和膜蛋白普遍减少。这些观察结果表明,震颤小鼠的病理生物学扩展到了肝脏,其中精神鞘氨醇诱导的TNF-α和IL-6损害了过氧化物酶体的结构和功能。